Home About us Editorial board Ahead of print Current issue Search Archives Submit article Instructions Subscribe Contacts Login 

 Table of Contents  
Year : 2007  |  Volume : 3  |  Issue : 2  |  Page : 86-91

Antitumor activity of 4-Arylcoumarins from endophytic Streptomyces aureofaciens CMUAc130

1 Department of Microbiology, Faculty of Science, Silpakorn University, Nakorn Pathom - 73000, Thailand
2 School of Life Sciences, Xiamen University, Xiamen - 361005, China
3 Kunming Institute of Botany, The Chinese Academy of Sciences, Kunming - 650204, China
4 Department of Biology, Faculty of Science, Chiang Mai University, Chiang Mai - 50200, Thailand

Correspondence Address:
Thongchai Taechowisan
Department of Microbiology, Faculty of Science, Silpakorn University, Nakorn Pathom - 73000
Login to access the Email id

Source of Support: None, Conflict of Interest: None

DOI: 10.4103/0973-1482.34685

Rights and Permissions
 > Abstract 

In a search for antitumor agents, we carried out a screening of 4-arylcoumarins isolated from endophytic Streptomyces aureofaciens CMUAc130, by examining their possible inhibitory effect on the growth of s.c. transplanted Lewis lung carcinoma (LLC) in BDF-1 mice by intraperitoneal (i.p.) administration. The 4-arylcoumarins showed antitumor activity with T/C values of 80.8 and 50.0% at doses of 1 and 10 mg/kg of 5,7-dimethoxy-4-p-methoxylphenylcoumarin treatment, respectively and 81.5 and 44.9% at doses of 1 and 10 mg/kg of 5,7-dimethoxy-4-phenylcoumarin treatment, respectively, compared to adriamycin, which was used a positive control, with T/C value of 55.9% at 2 mg/kg. Furthermore, we investigated the possible effects of these compounds on expression of the bcl-2 and Bax oncoproteins in A427, a human lung cancer cell lines. The cells were cultured in vitro for 24h in RPMI 1640 with 1.5% (v/v) ethanol, 100 µg/ml 5,7-dimethoxy-4-p-methoxylphenylcoumarin or 5,7-dimethoxy-4-phenylcoumarin. Viability was determined by an MTT assay. Total protein was extracted from cell lysates and the bcl-2 and Bax oncoproteins were identified. Western blotting showed a decrease in bcl-2 and an increase in Bax in A427 cell cultured with 5,7-dimethoxy-4-p-methoxylphenylcoumarin or 5,7-dimethoxy-4-phenylcoumarin. We conclude that 5,7-dimethoxy-4-phenylcoumarin is a more potent inhibitor of cell proliferation than 5,7-dimethoxy-4-p-methoxylphenylcoumarin and has more marked effects on oncoprotein expression.

Keywords: 4-arylcoumarins, antitumor activity, endophytic streptomyces, Lewis lung carcinoma

How to cite this article:
Taechowisan T, Lu C, Shen Y, Lumyong S. Antitumor activity of 4-Arylcoumarins from endophytic Streptomyces aureofaciens CMUAc130. J Can Res Ther 2007;3:86-91

How to cite this URL:
Taechowisan T, Lu C, Shen Y, Lumyong S. Antitumor activity of 4-Arylcoumarins from endophytic Streptomyces aureofaciens CMUAc130. J Can Res Ther [serial online] 2007 [cited 2022 Nov 29];3:86-91. Available from: https://www.cancerjournal.net/text.asp?2007/3/2/86/34685

Table 3: Inhibition of proliferation of A427 cells cultured for 24 h in RPMI 1640 with 1.5% (v/v) ethanol (control), 100 ěg/ml coumarins

Click here to view
Table 3: Inhibition of proliferation of A427 cells cultured for 24 h in RPMI 1640 with 1.5% (v/v) ethanol (control), 100 ěg/ml coumarins

Click here to view
Table 2: Inhibitory effects of coumarins on s.c. transplanted Lewis lung carcinoma cells

Click here to view
Table 2: Inhibitory effects of coumarins on s.c. transplanted Lewis lung carcinoma cells

Click here to view
Table 1: Normalization of densitometric values of bcl-2 and Bax oncoproteins with respect to âtubulin (n = 3)

Click here to view
Table 1: Normalization of densitometric values of bcl-2 and Bax oncoproteins with respect to âtubulin (n = 3)

Click here to view

 > Introduction Top

Coumarins are synthesized by numerous green plant species [1] and by some bacteria and fungi and they have low toxicity in human. They exhibit antibacterial, [2] antioxidant, [3] anticancer [4] and antiallergy activities. [5] Recent report describe that 4-hydroxycoumarin derivatives have antiproliferative effects against five human tumor cell lines (A549 (nonsmall cell lung carcinoma), SK-OV-3 (ovarian carcinoma), SK-MEL-2 (melanoma), XF498 (CNS carcinoma) and HCT-15 (codon carcinoma)) [6] and act directly on malignant cells reducing tumor size [7] but the mechanisms involved have not been described in detail. In a preliminary communication, we have reported the isolation of 5,7-dimethoxy-4-p-methoxylphenylcoumarin and 5,7-dimethoxy-4-phenylcoumarin from the crude extraction of Streptomyces aureofaciens CMUAc130 culture. [8] In the course of searching for the antitumor agent from these compounds, we investigated the antitumor effect of these compounds on Lewis lung carcinoma (LLC) cells in vivo and the biochemical mechanisms of bcl-2 and BAX expression in human lung cancer cell lines, A427 (adenocarcinoma). The 26 kDa membrane protein bcl-2 may extend the life of the cell by blocking free oxygen radicals, inhibiting cytochrome c released from mitochondria and suppressing caspase-9 activation. [9] This would cause oncogenic mutations to accumulate in the normal cell, which may facilitate carcinogenesis. Over-expression of bcl-2 may also delay the response of tumor cells to applied therapies. [10],[11] Over expression of Bax, a 23 kDa membrane protein, results in the formation of homodimers and facilitates the production of channels in the outer mitochondrial membrane and loss of mitochondrial membrane potential that allows cytochrome c release. In the cytoplasm, cytochrome c binds Apaf-1 in the presence of ATP and activates inductor caspase-9, which in turn activates executor caspases that cleave death substrates, key cytoplasmic and nuclear proteins. [12],[13] Coumarins bind nonspecifically to cell surface structures like receptors. They function as first messengers, inducing changes in signal transduction pathways involving GTP-binding proteins. Therefore, coumarins are able to mediate modifications of downstream signals leading to growth (e.g. cell-growth inhibition) and possibly decreasing bcl-2 expression. It has been shown that other coumarin derivatives exert their antiproliferative effect by modulation of the cell cycle, [14] although the mechanism responsible for this activity has not yet been clarified. Accordingly, our findings indicate that 5,7-dimethoxy-4-p-methoxylphenylcoumarin and 5,7-dimethoxy-4-phenylcoumarin may display antitumor activity by inhibiting progression through the G0/G1 and S phases of the cell cycle. The Bcl-2 family regulates the fate of normal cells and comprises approximately eight members divided into two groups. Group 1 includes bcl-2 and bcl-XL, which has a survival action; group 2 includes Bax, Bad, Bid and Bak, which has a survival inhibition action. [9] The susceptibility of a cell to adverse stimuli is determined in most cases by the dimerization of Bcl-2 family proteins through the highly conserved BH1, BH2 and BH3 domains. [15],[16] The loss of survival inhibition Bax proteins or overexpression of the proteins that block them, results in resistance to treatment; this was observed by Pezzella et al. , [17] during a study of apoptosis inhibition in non-small cell lung carcinoma. It has also been reported that an increase in Bax expression sensitizes cells to chemotherapeutic agents. [18] The purpose of the present study was to determine whether 5,7-dimethoxy-4-p-methoxylphenylcoumarin and 5,7-dimethoxy-4-phenylcoumarin alter the tumor proliferation in vivo and the expression of the Bcl-2 oncoprotein family.

 > Materials and Methods Top

Organisms and compounds

Streptomyces aureofaciens CMUAc130 was isolated from the root tissues of Zingiber officinale by surface-sterilization technique. [19],[20] Identification of the isolate to species level was based on morphological, cultural, physiological and biochemical characteristics and also 16S rDNA gene sequencing as described by Taechowisan and Lumyong. [19],[20] Solid medium for sporulation used in this study was International Streptomyces Project Medium 4 (ISP-4) and the liquid medium used for fermentation was ISP-2. [21] The large scale fermentation, crude extraction and purification of the compounds, was carried on the methods of Taechowisan et al . [8]

Structure elucidation of the compounds

The structures of the active compounds have been identified using NMR and mass spectral data. The melting point of the compounds was determined on a Buchi-540 melting point apparatus. Optical rotations were measured on a Perkin-Elmer 241 polarimeter, IR spectra on a Perkin-Elmer 1 spectrometer, 1 H and 13 C NMR spectra on a Bruker DRX 500 spectrometer and EI-MS and FAB-MS respectively on a Hewlett-Packard 5989 B and a Finnigan/Thermo Quest Mat 95 XL mass spectrometer.

Tumor transplantation: Lewis lung carcinoma (LLC, mouse) cells were maintained as monolayer cultures in RPMI 1640 medium supplemented with 10% fetal bovine serum, sodium bicarbonate (2 g), pencillin G (100,000 units) and streptomycin (100 mg). A suspension of 2.5×10 5 cells/0.1 ml/mouse in 0.85% NaCl solution was inoculated subcutaneously into the left flank of female BDF-1 mice (four weeks of age, weighing 14 to 16 g) for the subcutaneous tumor assay. [22] The purified compounds, 5,7-dimethoxy-4-p-methoxylphenylcoumarin and 5,7-dimethoxy-4-phenylcoumarin were suspended in 0.85% NaCl solution (0.1 ml) and administered intraperitoneally (i.p) once daily for consecutive day 1 to day 14 to mice. Control animals were given 0.1 ml of 0.85% NaCl solution by i.p. injection. Each treatment consisted of three replicates. The experiment was repeated twice. Tumors were measured each alternate day using a vernier caliper from the initiation of treatment to the time when grosses ulceration of the tumor was developed in control mice. The tumor size was calculated as:

Tumor vol. (mm 3 ) = 0.5×a×b 2 Where a is the longest diameter and b is the shortest diameter. [22] The effects by treatments were represented as follows:

T/C (%) = (mean value of a treated group/mean value of a control group) × 100. The significance of differences between the experimental group was calculated by Dunnett's t test. P <0.05 was considered significant. Expression of bcl-2 and Bax in cancer cell line by purified compounds. Human lung carcinoma cell line used was A427 (adenocarcinoma) ATCC (American Type Culture Collection, MA, USA). For controls we used HeLa cells, which express the Bax oncoprotein and isolated human peripheral blood lymphocytes from healthy donors, which express the bcl-2 protein. These cells were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum and maintained at 37˚C in a humidified 5% CO2 atmosphere until 80-85% confluence was reached. They were harvested by using 0.25% Trypsin and their viability was determined by Trypan blue exclusion. A427 cells were seeded at a viable density of 1 × 10 7 in 10 ml RPMI 1640-supplemented medium. Untreated cell lines or cells in 1.5% (v/v) ethanol (solvent), 100 µg/ml 5,7-dimethoxy-4-p-methoxylphenylcoumarin or 100 µg/ml 5,7-dimethoxy-4-phenylcoumarin, were cultured for 24 h. Each culture was photographed at 200x with a Nikon H-III Power, FDX-35 inverted microscope to reveal ethanol and purified compounds-induced morphological changes.

MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay

Changes in the number of viable cells were determined by the MTT assay. [23] Briefly, cells were seeded a viable density of 5 × 10 3 /100 µl RPMI/ well in a 96 well microplate. Untreated cell lines or cells in 1.5% (v/v) ethanol (solvent and control), 100 µg/ml 5,7-dimethoxy-4-p-methoxylphenylcoumarin or 5,7-dimethoxy-4-phenylcoumarin, [8] were cultured for 24h and the number of viable cells was estimated by adding 20 µl/well of MTT (5 mg/ml PBS) for 4h, enabling viable cell mitochondrial succinate dehydrogenase to reduce MTT to purple formazan crystals. The medium was aspirated and 100 µl DMSO/well was added. The crystals were dissolved in DMSO and the absorbance at 570 nm was measured with an ELISA reader (Bio-Tec Instrum. EL311). Three separate experiments were conducted and each was performed in triplicate. Statgraphic software was used to determine probability values (p) and statistically significant differences (*). The percentage inhibition of cell viability was measured using the formula:

%IC = [(1- cells with compound/cells in solvent)100] Protein isolation and Western blot analysis. Cells from each culture were lysed to extract total proteins by the TRIZOL method. [24] Brielfly, RNA and DNA were removed from the cell lysates and total proteins were precipitated from the final supernatant by adding 0.2 ml isopropanol per milliliter of TRIZOL used initailly. The precipitate was washed with 0.2% guanidine in 95% ethanol, then dissolved in 1% SDS and stored at -20˚C until it was analyzed. The total protein concentration was determined by the Petersen method. [25] Samples of total protein (30 µg/well) were separates on 10% SDS-PAGE, transferred to nylon membranes (Hybond-C Amersham Life Technologies) and incubated with the first antibody: anti-bcl-2(monoclonal antibody clone Bcl-2-100, labeling 26 kDa bands), anti-Bax (monoclonal antibody clone 2D2, labeling 23 kDa bands) (both from Zymed Laboratories, Inc. San Francisco, CA USA) or anti-β -tubulin (Monoclonal antibody clone Tub 2.1, labeling 55 kDa bands) (Sigma-Aldrich Inc, St. Louis MO, USA). The second (antimouse) antibody tagged with alkaline phosphatase was added with a 5-bromo-4-chloro-5-indolyl phosphate toluidine/nitroblue tetrazolium substrate as a color development reagent (Roche Diagnostics Co, Molecular Biochemical, Indianapolis, IN, USA). The band intensities were quantified by densitometry using RFLP scan software (Scanalytics, v. 2.1, 1994; Scanalytics, a division of CSPIMA, USA). The bcl-2 and Bax band values were normalized to the β -tubulin value (value = 1). Western blotting assays were repeated at least three times and a representative gel is included as [Figure - 1]. Data in [Table - 1] indicate the mean and standard deviation of three normalized densitometry values.

 > Results and Discussion Top

We have studied physiological roles of 5,7-dimethoxy-4-p-methoxylphenylcoumarin and 5,7-dimethoxy-4-phenylcoumarin for the development of biologically active substances. [8] Although there are many coumarin derivatives have special ability to scavenge reactive oxygen species-free radicals, such as hydroxyl radicals, superoxide radicals or hypochlorous acid and to influence processes involving free radical-injury. [26] They have also been found to inhibit lipid peroxidation and to possess vasorelaxant, [27] anticoagulant activities, [28] antiinflammatory/antioxidant activity. [29],[30],[31] Moreover they showed potent in vitro antitumor activity. [6] We selected 5,7-dimethoxy-4-p-methoxylphenylcoumarin and 5,7-dimethoxy-4-phenylcoumarin with a potent antitumor effect on LLC cells in vivo and the biochemical mechanisms of bcl-2 and BAX expression in A427. [Figure - 2] shows chemical structures of 5,7-dimethoxy-4-p-methoxylphenylcoumarin and 5,7-dimethoxy-4-phenylcoumarin. The purified compounds, 5,7-dimethoxy-4-p-methoxylphenylcoumarin and 5,7-dimethoxy-4-phenylcoumarin were evaluated for antitumor activity against subcutaneous transplanted LLC cells, in vivo . These compounds, when given intraperitoneally once daily for the 14 days at dosages of 1 and 10 mg/kg, inhibited tumor growth with T/C values of 80.8 and 50.0% for 5,7-dimethoxy-4-p-methoxylphenylcoumarin treatment, respectively and 81.5 and 44.9% for 5,7-dimethoxy-4-phenylcoumarin treatment, respectively, compared with that of the control [Figure - 3]. When the sample was intraperitoneally administrated once daily for 14 days, the body weights continued to until the fifth day after injection, following gradual increase and weighing almost same as body weights in control group [Table - 2]. Coumarins have been studied for their binding properties to tumor cell membranes in vitro , for their ability to inhibit cell proliferation in cells dependent on the over-expression of oncogenes. In order to measure the interaction, intact target cells were radioactively labeled with coumarin. Metastatic breast carcinoma cells showed high-affinity binding properties, where an adenocarcinoma raised from a human colorectal primary tumor exhibited low affinity to coumarin. The binding properties of coumarin to target cells are enigmatic, possibly due to biological variability. [32] Diffusion of coumarin molecules through the cell is determined by the phospholipid composition of the plasma membrane. Thus, the level of affinity of coumarins for membrane phospholipids constitutes the driving force that allows the entry of coumarins into the cell. [Figure - 4]a shows the untreated A427 cell line culture, while [Figure - 4]b shows this culture treated with ethanol. In both cases, the cells grew separately, adopting irregular forms with cytoplasmic elongations. [Figure - 4]c and d is the culture treated with 5,7-dimethoxy-4-p-methoxylphenylcoumarin and 5,7-dimethoxy-4-phenylcoumarin, respectively. Small, round cells tending to group together were observed in both these cultures and are particularly prevalent in [Figure - 4]d. No morphological changes were induced by ethanol. An MTT assay was performed to determine the inhibition of cell proliferation in the two cell lines cultured for 24-h with 100 µg/ml 5,7-dimethoxy-4-p-methoxylphenylcoumarin or 5,7-dimethoxy-4-phenylcoumarin or with 1.5% (v/v) ethanol (solvent control). [Table - 3] shows the results processed by the SPSS 9.05 software. The 5,7-dimethoxy-4-phenylcoumarin inhibited A427 cell growth by 42%, while 5,7-dimethoxy-4-p-methoxylphenylcoumarin inhibited it by 64% (* P <0.05). These results suggest that 5,7-dimethoxy-4-p-methoxylphenylcoumarin is more active than 5,7-dimethoxy-4-phenylcoumarin inhibited, probably due to the methoxyl- group at the 4-carbon position on the phenyl of these compounds. The effects of 5,7-dimethoxy-4-p-methoxylphenylcoumarin, 5,7-dimethoxy-4-phenylcoumarin and ethanol solvent on bcl-2 and Bax oncoprotein expression after 24-h in the A427 cultures are shown in [Figure - 1]. Bcl-2 expression (survival action) was decreased in the A427 cells treated with 5,7-dimethoxy-4-p-methoxylphenylcoumarin and 5,7-dimethoxy-4-phenylcoumarin and Bax expression was slightly increased with these coumarins. Western blotting assays were repeated at least three times and a representative gel is included as [Figure - 1]. The densitometric values for the bcl-2, Bax and β -tubulin bands are given in [Table - 1]; the mean and SD of three normalized bcl-2 and Bax values with respect to β -tubulin (value = 1) are shown. Research in our laboratory indicates that the proliferation capacity of a human adenocarcinoma cell line (A427), overexpressing the bcl-2 oncogene, can be reduced when incubated with 5,7-dimethoxy-4-p-methoxylphenylcoumarin and 5,7-dimethoxy-4-phenylcoumarin. Using Western blotting, we have demonstrated with these cells that, when coumarins are present for 24-h in the cell culture medium, the bcl-2 protein is decreased, which raises the intriguing possibility that further intracellular growth signal processing is almost abandoned, leading to inhibition of cell proliferation. These observations are in agreement with the well-known antiproliferative effect of coumarins on malignant cell lines, decreasing bcl-2 expression and its survival action.

There is an urgent need for agents with minimal side effects but with significant antitumor activity for particular cancers. 5,7-dimethoxy-4-phenylcoumarin might be effective in preventing or delaying the formation of metastases; they are candidates for adjuvant treatment strategies to delay metastases. Both 5,7-dimethoxy-4-p-methoxylphenylcoumarin and 5,7-dimethoxy-4-phenylcoumarin exhibit low toxicity in normal cells and their effect on malignant cells recommends them both as chemopreventatives and for use in combined antitumor treatment.

 > Acknowledgement Top

We are grateful to Prof. Pittaya Tantiwachwuttikul for isolation the pure compounds. This research project was funded by Thailand Research Fund MRG4980137 and Faculty of Science, Silpakorn University, Nakorn Pathom, Thailand.

 > References Top

1.Murray RD, Mιndez J, Brown SA. The natural coumarins. John Wiley: Chichester; 1982.  Back to cited text no. 1
2.Inoue Y, Kondo H, Taguchi M, Jinbo Y, Sakamoto F, Tsukamoto G. Synthesis and antibacterial activity of thiazolopyrazine-incorporated tetracyclic quinolone antibacterials. J Med Chem 1994;37:586-92.  Back to cited text no. 2
3.Jamkhandi PS, Rajagopal S. Synthesis of 4-hydroxy-l-thiacoumarins. Arch Pharm Ber Dtsch Pharm Ges 1967;300:561-6.  Back to cited text no. 3
4.Badawi MM, Handa SS, Kinghorn AD, Cordell GA, Farnsworth NR. Plant anticancer agents XXVII: Antileukemic and cytotoxic constituents of Dirca occidentalis (Thymelaeaceae). J Pharm Sci 1983;72:1285-7.  Back to cited text no. 4
5.Buckle DR, Cantello BC, Smith H, Spicer BA. 2-cyano-1,3-dicarbonyl compounds with antiallergic activity. J Med Chem 1977;20:265-9.  Back to cited text no. 5
6.Jung JC, Lee JH, Oh S, Lee JG, Park OS. Synthesis and antitumor activity of 4-hydroxycoumarin derivatives. Bioorg Med Chem Lett 2004;14:5527-31.  Back to cited text no. 6
7.Kawaii S, Tomono Y, Ogawa K, Sugiura M, Yano M, Yoshizawa Y. The antiproliferative effect of coumarins on several cancer cell lines. Anticancer Res 2001;21:917-23.  Back to cited text no. 7
8.Taechowisan T, Lu C, Shen Y, Lumyong S. Secondary metabolites from endophytic Streptomyces aureofaciens CMUAc130 and their antifungal activity. Microbiology 2005;151:1691-5.  Back to cited text no. 8
9.Yang J, Liu X, Bhalla K, Kim CN, Ibrado AM, Cai J, et al . Prevention of apoptosis by bcl-2: Release of cytochrome c from mitochondria blocked. Science 1997;275:1129-32.  Back to cited text no. 9
10.Reed JC, Miyashita T, Takayama S, Wang HG, Sato T, Krajewiski S, et al . BCL-2 family proteins regulators of cell death involved in the pathogenesis of cancer and resistance to therapy. J Cell Biochem 1996;60:23-32.  Back to cited text no. 10
11.Meyn RE, Stephens LC, Milas L. Programmed cell death and radioresistance. Cancer Metastasis Rev 1996;15:119-31.  Back to cited text no. 11
12.Yoshida H, Kong YY, Yoshida R, Elia AJ, Hakem A, Hakem R, et al . Apaf-1 is required for mitochondrial pathways of apoptosis and brain development. Cell 1998;94:739-50.  Back to cited text no. 12
13.Yamaguchi H, Wang HG. The protein kinase PKB/Akt regulates cell survival and apoptosis by inhibiting Bax conformational change. Oncogene 2001;20:7780-6.  Back to cited text no. 13
14.Kahn J, Preis P, Waldman F, Tseng A. Coumarin modulates the cell-cycle progression of an MTV-EJras cell line. J Cancer Res Oncol 1994;120:S19-22.  Back to cited text no. 14
15.Sedlak TW, Oltvai ZN, Yank E, Wang K, Boise LH, Thompson CB, et al . Multiple Bcl-2 family members demonstrate selective dimerizations with Bax. Proc Natl Acad Sci USA 1995;92:7834-8.  Back to cited text no. 15
16.Zha H, Aimι-Sempι C, Sato T, Reed JC. Proapoptotic protein Bax heterodimerizes with bcl-2 and homodimerizes with Bax via a novel domain (BH3) distinct from BH1 and BH2. J Biol Chem 1996;271:7440-4.  Back to cited text no. 16
17.Pezzella F, Turley H, Kuru I, Tungekar MH, Pierce CB, Harris A, et al . Bcl-2 protein in non-small-cell lung carcinoma. N Engl J Med 1993;329:690-4.  Back to cited text no. 17
18.Guo B, Cao S, Toth K, Azrac RG, Rustum YM. Overexpression of Bax enhances antitumor activity of chemotherapeutic agents in human head and neck squamous cell carcinoma. Clin Cancer Res 2000;6:718-24.  Back to cited text no. 18
19.Taechowisan T, Lumyong S. Activity of endophytics actinomycetes from roots of Zingiber officinale and Alpinia galanga against phytopathogenic fungi. Ann Microbiol 2003;53:291-8.  Back to cited text no. 19
20.Taechowisan T, Peberdy JF, Lumyong S. Isolation of endophytic actinomycetes from selected plants and their antifungal activity. World J Microbiol Biotechnol 2003;19:381-5.  Back to cited text no. 20
21.Shirling EB, Gottlieb D. Methods for characterization of Streptomyces species. Int J Syst Bacteriol 1966;16:313-40.  Back to cited text no. 21
22.Kato T, Sato K, Kakinuma H, Matsuda Y. Enhanced suppression of tumor growth by combination of angiogenesis inhibitor O-(chloroacetyl-carbamoyl) fumagillol (TNP-470) and cytotoxic agents in mice. Cancer Res 1994;54:5143-7.  Back to cited text no. 22
23.Mosmann T. Rapid colorimetric assay for cellular growth and survival: Applications to proloferation and cytotoxicity assay. J Immunol 1983;65:55-63.  Back to cited text no. 23
24.Chomczynski P, Sacchi N. Single step method of RNA isolation by acid guanidinium thiocyanate phenol chloroform extraction. Anal Biochem 1986;162:156-9.  Back to cited text no. 24
25.Peterson GL. Determination of total protein. Methods Enzymol 1983;81:95-119.  Back to cited text no. 25
26.Mora A, Paya M, Rios JL, Alcaraz MJ. Structure-activity relationships of polymethoxyflavones and other flavonoids as inhibitors of non-enzymic lipid peroxidation. Biochem Pharmacol 1990;40:793-7.  Back to cited text no. 26
27.Hoult JR, Paya M. Pharmacological and biochemical actions of simple coumarins: Natural products with therapeutic potential. Gen Pharmacol 1996;27:713-22.  Back to cited text no. 27
28.Visser LE, Trienekens PH, De Smet PA, Vulto AG, Hofman A, van Duijn CM. et al . Patients with an ApoE epsilon4 allele require lower doses of coumarin anticoagulants. Pharmacogenet Genomics 2000;15:69-74.  Back to cited text no. 28
29.Nicolaides DN, Gautam DR, Litinas KE, Hadjipavlou-Litina DJ, Fylaktakidou, KC. Synthesis and evaluation of the antioxidant and antiinflammatory activities of some benzo[l]khellactone derivatives and analogues. Eur J Med Chem 2004;39:323-32.  Back to cited text no. 29
30.Kontogiorgis CA, Hadjipavlou-Litina DJ. Synthesis and biological evaluation of novel coumarin derivatives with a 7-azomethine linkage. Bioorg Med Chem Lett 2004;14:611-4.  Back to cited text no. 30
31.Fylaktakidou KC, Hadjipavlou-Litina DJ, Litinas KE, Nicolaides DN. Natural and synthetic coumarin derivatives with anti-inflammatory/antioxidant activities. Curr Pharm Des 2004;10:3813-33.  Back to cited text no. 31
32.Zδnker KS. Coumarin(s) interface between cell growth inhibition, expression of oncogenes, tumour cell locomotion and cell-to-cell communication. Irish Coll Phys Surg 1993;22:51-5.  Back to cited text no. 32


  [Figure - 1], [Figure - 2], [Figure - 3], [Figure - 4]

  [Table - 1], [Table - 2], [Table - 3]

This article has been cited by
1 Plants and endophytes – a partnership for the coumarin production through the microbial systems
Chandrashekar Srinivasa, Govindappa Mellappa, Shashank M. Patil, Ramith Ramu, Bhargav Shreevatsa, Chandan Dharmashekar, Shiva Prasad Kollur, Asad Syed, Chandan Shivamallu
Mycology. 2022; : 1
[Pubmed] | [DOI]
2 Therapeutic potentials of endophytes for healthcare sustainability
Ayodeji O. Falade, Kayode E. Adewole, Temitope C. Ekundayo
Egyptian Journal of Basic and Applied Sciences. 2021; 8(1): 117
[Pubmed] | [DOI]
3 Discovery of Novel 3,4-Dichloroisothiazole-Containing Coumarins as Fungicidal Leads
You Lv, Hanlu Liu, Lifan Wang, Kun Li, Wei Gao, Xiaoyu Liu, Liangfu Tang, Tatiana A. Kalinina, Tatiana V. Glukhareva, Zhijin Fan
Journal of Agricultural and Food Chemistry. 2021; 69(14): 4253
[Pubmed] | [DOI]
4 Endophytic Microbial Diversity: A New Hope for the Production of Novel Anti-tumor and Anti-HIV Agents as Future Therapeutics
Aditya Banyal, Vikram Thakur, Rahul Thakur, Pradeep Kumar
Current Microbiology. 2021; 78(5): 1699
[Pubmed] | [DOI]
5 Design, synthesis and fungicidal activity of 3,4-dichloroisothiazolocoumarin-containing strobilurins
You Lv, Kun Li, Wei Gao, Zesheng Hao, Weibo Wang, Xiaoyu Liu, Liangfu Tang, Zhijin Fan
Molecular Diversity. 2021;
[Pubmed] | [DOI]
6 Alkynoates as Versatile and Powerful Chemical Tools for the Rapid Assembly of Diverse Heterocycles under Transition-Metal Catalysis: Recent Developments and Challenges
Imtiaz Khan, Aliya Ibrar, Sumera Zaib
Topics in Current Chemistry. 2021; 379(1)
[Pubmed] | [DOI]
7 Evaluation of the anticarcinogenic potential of the endophyte, Streptomyces sp. LRE541 isolated from Lilium davidii var. unicolor (Hoog) Cotton
Aiai Ma, Kan Jiang, Bin Chen, Shasha Chen, Xinge Qi, Huining Lu, Junlin Liu, Xuan Zhou, Tan Gao, Jinhui Li, Changming Zhao
Microbial Cell Factories. 2021; 20(1)
[Pubmed] | [DOI]
8 Beneficial Relationships Between Endophytic Bacteria and Medicinal Plants
Wei Wu, Wenhua Chen, Shiyu Liu, Jianjun Wu, Yeting Zhu, Luping Qin, Bo Zhu
Frontiers in Plant Science. 2021; 12
[Pubmed] | [DOI]
9 Microbial endophytes: an untapped resource with antitumor and anti-microbial properties
Preetinder Kaur, Gurpreet Kaur
The Applied Biology & Chemistry Journal. 2020; : 9
[Pubmed] | [DOI]
10 Diversified Thiazole Substituted Coumarins and Chromones as Non- Cytotoxic ROS and NO Inhibitors
Uzma Salar, Khalid Mohammed Khan, Almas Jabeen, Shafquat Hussain, Aisha Faheem, Farwa Naqvi, Shahnaz Perveen
Letters in Drug Design & Discovery. 2020; 17(5): 547
[Pubmed] | [DOI]
11 Elucidating Mechanisms of Endophytes Used in Plant Protection and Other Bioactivities With Multifunctional Prospects
Ayomide Emmanuel Fadiji, Olubukola Oluranti Babalola
Frontiers in Bioengineering and Biotechnology. 2020; 8
[Pubmed] | [DOI]
12 Diverse endophytic Streptomyces species with dynamic metabolites and their meritorious applications: a critical review
Reshma Ayswaria, Vineeth Vasu, Rijesh Krishna
Critical Reviews in Microbiology. 2020; 46(6): 750
[Pubmed] | [DOI]
13 Study of the Component Structure of the Metabolites of Bacteria Nocardiopsis umidischolae in the Search for Eco-Friendly Plant Protection Agents
L. N. Grigoryan, Yu. V. Bataeva, E. D. Andreeva, D. Kh. Zakar’yaeva, Z. O. Turaeva, S. V. Antonova
Russian Journal of General Chemistry. 2020; 90(13): 2531
[Pubmed] | [DOI]
14 Synthesis and biological evaluation of 4 arylcoumarin analogues as tubulin-targeting antitumor agents
Peggoty Mutai, Gilles Breuzard, Alessandra Pagano, Diane Allegro, Vincent Peyrot, Kelly Chibale
Bioorganic & Medicinal Chemistry. 2017; 25(5): 1652
[Pubmed] | [DOI]
15 Visible-Light-Promoted Dual C–C Bond Formations of Alkynoates via a Domino Radical Addition/Cyclization Reaction: A Synthesis of Coumarins
Shangbiao Feng, Xingang Xie, Weiwei Zhang, Lin Liu, Zhuliang Zhong, Dengyu Xu, Xuegong She
Organic Letters. 2016; 18(15): 3846
[Pubmed] | [DOI]
16 Silver-Mediated Radical Cyclization of Alkynoates and a-Keto Acids Leading to Coumarins via Cascade Double C–C Bond Formation
Kelu Yan, Daoshan Yang, Wei Wei, Fen Wang, Yuanyuan Shuai, Qiannan Li, Hua Wang
The Journal of Organic Chemistry. 2015; 80(3): 1550
[Pubmed] | [DOI]
17 Antioxidant and antitumor activities of 4-arylcoumarins and 4-aryl-3,4-dihydrocoumarins
Keyun Zhang,Weixian Ding,Jie Sun,Bin Zhang,Fujiao Lu,Ren Lai,Yong Zou,Gabriel Yedid
Biochimie. 2014;
[Pubmed] | [DOI]
18 An efficient synthesis of some new 3-bipyridinyl substituted coumarins as potent antimicrobial agents
Hemali B. Lad,Rakesh R. Giri,D.I. Brahmbhatt
Chinese Chemical Letters. 2013; 24(3): 227
[Pubmed] | [DOI]
19 Isolation of endophytic bacteria producing the anti-cancer alkaloid camptothecine from Miquelia dentata Bedd. (Icacinaceae)
Shweta, S. and Bindu, J.H. and Raghu, J. and Suma, H.K. and Manjunatha, B.L. and Kumara, P.M. and Ravikanth, G. and Nataraja, K.N. and Ganeshaiah, K.N. and Uma Shaanker, R.
Phytomedicine. 2013; 20(10): 913-917
20 Antitumor activity of bacterial exopolysaccharides from the endophyte bacillus amyloliquefaciens sp.isolated from ophiopogon japonicus
Chen, Y.-T. and Yuan, Q. and Shan, L.-T. and Lin, M.-A. and Cheng, D.-Q. and Li, C.-Y.
Oncology Letters. 2013; 5(6): 1787-1792
21 An efficient synthesis of some new 3-bipyridinyl substituted coumarins as potent antimicrobial agents
Lad, H.B. and Giri, R.R. and Brahmbhatt, D.I.
Chinese Chemical Letters. 2013; 24(3): 227-229
22 In vitro growth inhibitory activity of Prosopis cineraria leaves in MCF-7 breast cancer cell line
Sumathi, S. and Dharani, B. and Sivaprabha, J. and Sonia Raj, K. and Padma, P.R.
International Journal of Research in Pharmaceutical Sciences. 2013; 4(1): 40-44
23 Synthesis, biological evaluation and molecular docking studies of 3-(triazolyl)-coumarin derivatives: Effect on inducible nitric oxide synthase
Stefani, H.A. and Gueogjan, K. and Manarin, F. and Farsky, S.H.P. and Zukerman-Schpector, J. and Caracelli, I. and Pizano Rodrigues, S.R. and Muscará, M.N. and Teixeira, S.A. and Santin, J.R. and MacHado, I.D. and Bolonheis, S.M. and Curi, R. and Vinolo, M.A.
European Journal of Medicinal Chemistry. 2012; 58: 117-127
24 Coumarin-based benzopyranone derivatives induced apoptosis in human lung (A549) cancer cells
Musa, M.A. and Badisa, V.L.D. and Latinwo, L.M. and Patterson, T.A. and Owens, M.A.
Anticancer Research. 2012; 32(10): 4271-4276
25 Endophytic streptomycetes from plants with novel green chemistry: Review
Joseph, B. and Sankarganesh, P. and Edwin, B.T. and Raj, S.J.
International Journal of Biological Chemistry. 2012; 6(2): 42-52
26 Characterization of an endophytic whorl-forming streptomyces from Catharanthus roseus stems producing polyene macrolide antibiotic
Rakotoniriana, E.F. and Chataigné, G. and Raoelison, G. and Rabemanantsoa, C. and Munaut, F. and El Jaziri, M. and Urveg-Ratsimamanga, S. and Marchand-Brynaert, J. and Corbisier, A.-M. and Declerck, S. and Quetin-Leclercq, J.
Canadian Journal of Microbiology. 2012; 58(5): 617-627
27 Alpha-glucosidase inhibitor activity and characterization of endophytic actinomycetes isolated from some Indonesian diabetic medicinal plants
Pujiyanto, S. and Lestari, Y. and Suwanto, A. and Budiarti, S. and Darusman, L.K.
International Journal of Pharmacy and Pharmaceutical Sciences. 2012; 4(SUPPL.1): 327-333
28 A novel source of bioactive compounds: Endophytic bacteria isolated from Centella asiatica
Rafat, A. and Philip, K. and Muniandy, S.
Journal of Pure and Applied Microbiology. 2012; 6(1): 11-20
29 Amelioration of oxidative stress induced by oxidative mutagens and COX-2 inhibitory activity of umbelliferone isolated from Glycyrrhiza glabra L
Kaur, P. and Kumar, M. and Singh, B. and Kumar, S. and Kaur, S.
Asian Pacific Journal of Tropical Biomedicine. 2012; 2(1 SUPPL.): S120-S126
30 Synthesis, biological evaluation and molecular docking studies of 3-(triazolyl)-coumarin derivatives: Effect on inducible nitric oxide synthase
Hélio A. Stefani,Karina Gueogjan,Flávia Manarin,Sandra H.P. Farsky,Julio Zukerman-Schpector,Ignez Caracelli,Sergio R. Pizano Rodrigues,Marcelo N. Muscará,Simone A. Teixeira,José R. Santin,Isabel D. Machado,Simone M. Bolonheis,Rui Curi,Marco A. Vinolo
European Journal of Medicinal Chemistry. 2012; 58: 117
[Pubmed] | [DOI]
31 Amelioration of oxidative stress induced by oxidative mutagens and COX-2 inhibitory activity of umbelliferone isolated from Glycyrrhiza glabra L
Prabhjit Kaur,Manish Kumar,Bikram Singh,Subodh Kumar,Satwinderjeet Kaur
Asian Pacific Journal of Tropical Biomedicine. 2012; 2(1): S120
[Pubmed] | [DOI]
32 Palladium-Catalyzed Oxidative Heck Coupling Reaction for Direct Synthesis of 4-Arylcoumarins Using Coumarins and Arylboronic Acids
Yaming Li, Zisong Qi, Huifeng Wang, Xinmei Fu, Chunying Duan
The Journal of Organic Chemistry. 2012; 77(4): 2053
[VIEW] | [DOI]
33 Characterization of an endophytic whorl-formingStreptomycesfromCatharanthus roseusstems producing polyene macrolide antibiotic
Erick Francisco Rakotoniriana,Gabrielle Chataigné,Guy Raoelison,Christian Rabemanantsoa,Françoise Munaut,Mondher El Jaziri,Suzanne Urveg-Ratsimamanga,Jacqueline Marchand-Brynaert,Anne-Marie Corbisier,Stéphane Declerck,Joëlle Quetin-Leclercq
Canadian Journal of Microbiology. 2012; 58(5): 617
[Pubmed] | [DOI]
34 Efficient synthesis and biological evaluation of 4-arylcoumarin derivatives
Jie Sun, Wei Xian Ding, Ke Yun Zhang, Yong Zou
Chinese Chemical Letters. 2011; 22(6): 667
[VIEW] | [DOI]
35 Biodiversity, bioactive natural products and biotechnological potential of plant-associated endophytic actinobacteria
Sheng Qin, Ke Xing, Ji-Hong Jiang, Li-Hua Xu, Wen-Jun Li
Applied Microbiology and Biotechnology. 2011; 89(3): 457
[VIEW] | [DOI]
36 Synthesis of hydrophilic and lipophilic 4-arylcoumarin phosphates
A. N. Selikhov,Y. B. Malysheva,A. V. Nyuchev,N. S. Sitnikov,E. A. Sharonova,A. S. Shavyrin,S. Combes,A. Yu. Fedorov
Russian Chemical Bulletin. 2011; 60(10): 2003
[Pubmed] | [DOI]
37 Synthesis of hydrophilic and lipophilic 4-arylcoumarin phosphates
Selikhov, A.N. and Malysheva, Y.B. and Nyuchev, A.V. and Sitnikov, N.S. and Sharonova, E.A. and Shavyrin, A.S. and Combes, S. and Fedorova, A.Yu.
Russian Chemical Bulletin. 2011; 60(10): 2003-2009
38 Need exploration of endophytic streptomyces - Review
Joseph, B. and Sankarganesh, P. and Justinraj, S. and Edwin, B.T.
Journal of Pure and Applied Microbiology. 2011; 5(1): 153-160
39 Synthesis of 4-(2-hydroxymethylaryl)coumarins
N. S. Sitnikov, A. S. Shavyrin, G. K. Fukin, I. P. Beletskaya, S. Combes, A. Yu. Fedorov
Russian Chemical Bulletin. 2010; 59(3): 626
[VIEW] | [DOI]
40 Chemical and functional diversity of natural products from plant associated endophytic fungi
Verma, V.C. and Kharwar, R.N. and Strobel, G.A.
Natural Product Communications. 2009; 4(11): 1511-1532
41 2-(Azidomethyl)arylboronic acids in the synthesis of coumarin-type compounds
Naumov, M.I., Nuchev, A.V., Sitnikov, N.S., Malysheva, Y.B., Shavyrin, A.S., Beletskaya, I.P., Gavryushin, A.E., (...), Fedorov, A.Yu.
Synthesis. 2009; 10(Z27508SS): 1673-1682
42 Synthesis and biological evaluation of polymethoxylated 4-heteroarylcoumarins as tubulin assembly inhibitor
Ganina, O.G., Daras, E., Bourgarel-Rey, V., Peyrot, V., Andresyuk, A.N., Finet, J.-P., Fedorov, A.Yu., (...), Combes, S.
Bioorganic and Medicinal Chemistry. 2008; 16(19): 8806-8812
43 Synthesis and biological evaluation of polymethoxylated 4-heteroarylcoumarins as tubulin assembly inhibitor
Olga G. Ganina,Etienne Daras,Véronique Bourgarel-Rey,Vincent Peyrot,Alexey N. Andresyuk,Jean-Pierre Finet,Alexey Yu. Fedorov,Irina P. Beletskaya,Sébastien Combes
Bioorganic & Medicinal Chemistry. 2008; 16(19): 8806
[Pubmed] | [DOI]


Similar in PUBMED
   Search Pubmed for
   Search in Google Scholar for
 Related articles
Access Statistics
Email Alert *
Add to My List *
* Registration required (free)

  >Abstract>Introduction>Materials and Me...>Results and Disc...>Acknowledgement>Article Figures>Article Tables
  In this article

 Article Access Statistics
    PDF Downloaded1090    
    Comments [Add]    
    Cited by others 43    

Recommend this journal