|Year : 2022 | Volume
| Issue : 2 | Page : 411-417
Effect of lentivirus-mediated peroxiredoxins 6 gene silencing on the phenotype of human gastric cancer BGC-823 cells
RunHong Mu1, YuPeng Li1, JiaYing Xing2, YanDong Li3, Rui Lin2, SiPing Ye2, YaoYue Zhang4, Han Mu5, Xiao Guo2, LiPing An2
1 Basic Medical College of Beihua University, Affiliated Hospital of Beihua University, Jilin, Jilin, P. R. China
2 Pharmacy College of Beihua University, Jilin, Jilin, P. R. China
3 Department of General Surgery, Affiliated Hospital of Beihua University, Jilin, Jilin, P. R. China
4 Stomatology College of Beihua University, Jilin, Jilin, P. R. China
5 Laboratory Medical College of Beihua University, Jilin, Jilin, P. R. China
|Date of Submission||07-Jul-2021|
|Date of Acceptance||30-Dec-2021|
|Date of Web Publication||20-May-2022|
East Campus of Beihua University, No. 3999 Binjiang East Road, Jiangnan Street, Fengman District, Jilin City, Jilin Province
P. R. China
East Campus of Beihua University, No. 3999 Binjiang East Road, Jiangnan Street, Fengman District, Jilin City, Jilin Province
P. R. China
Source of Support: None, Conflict of Interest: None
Aims: Peroxiredoxins (PRDX6) regulates the occurrence and progression of cancer. The aim of this study is to investigate the effect of PRDX6 knockdown on the biological behavior of human gastric cancer cell line BGC-823 cells.
Settings and Design: Research article.
Subjects and Methods: The differential expression of PRDX6 in gastric cancer and normal gastric tissues was tested by immunohistochemistry. Ribonucleic acid plasmid of PRDX6 gene was packaged using a lentivirus, and BGC-823 cells were transfected with the lentivirus to obtain a BGC-823 cell line in which the expression of PRDX6 was stably silenced.
Statistical Analysis Used: The proliferation activity of BGC-823 cells was detected using the cell counting kit-8 method. The effect of PRDX6 on the migration and invasion of BGC-823 cells was evaluated using the scratch test and Transwell assay, and the expression of related proteins was detected by western blot.
Results: The expression of PRDX6 in gastric cancer was significantly increased (P < 0.05). Compared with those in the untransfected and negative control groups. The proliferation, migration, and invasion of gastric cancer BGC-823 cells were significantly inhibited, and the apoptotic rates were significantly increased in the lentivirus-transfected (short hairpin-PRDX6) group. Western blot analysis showed that the expression of Bax protein increased, whereas that of proliferating cell nuclear antigen, Bcl-2, PI3K, phospho (p-Akt), and phosphorylated-mammalian target of rapamycin (mTOR) decreased significantly compared with that in WT and vector groups (P < 0.05).
Conclusion: The knockdown of PRDX6 gene expression in BGC-823 cells can inhibit the proliferation, migration, and invasion of gastric cancer cells and promote apoptosis, thereby affecting gastric cancer cells.
Keywords: Gastric cancer, migration, peroxiredoxins 6, proliferation
|How to cite this article:|
Mu R, Li Y, Xing J, Li Y, Lin R, Ye S, Zhang Y, Mu H, Guo X, An L. Effect of lentivirus-mediated peroxiredoxins 6 gene silencing on the phenotype of human gastric cancer BGC-823 cells. J Can Res Ther 2022;18:411-7
|How to cite this URL:|
Mu R, Li Y, Xing J, Li Y, Lin R, Ye S, Zhang Y, Mu H, Guo X, An L. Effect of lentivirus-mediated peroxiredoxins 6 gene silencing on the phenotype of human gastric cancer BGC-823 cells. J Can Res Ther [serial online] 2022 [cited 2022 Jul 7];18:411-7. Available from: https://www.cancerjournal.net/text.asp?2022/18/2/411/345522
RunHong Mu and YuPeng Li, these authors contributed to the work equally and should be regarded as co-first authors
| > Introduction|| |
Gastric cancer is one of the most common malignant tumors in the digestive tract. It is a serious threat to human health because of its high incidence, poor prognosis, and high mortality. The occurrence of gastric cancer is relatively occult; its initial symptoms are not obvious and most patients are diagnosed in middle and advanced stages, often accompanied by tumor metastasis, leading to the difficulty of its treatment., Therefore, it is very important to understand the molecular mechanism of gastric cancer occurrence and progression and identify diagnosis and treatment targets for the prevention and treatment of gastric cancer.
The survival of tumor cells is closely related to the state of intracellular and extracellular oxidative balance. Excessive production of reactive oxygen species (ROS) can cause the oxidation of cell membrane lipids, resulting in cellular damage, cardiovascular and cerebrovascular diseases, and cancer.,, Studies have indicated that the pathogenesis of gastric cancer may be the result of the combined action of multiple factors, such as oxidative stress, DNA mismatch, and loss of repair function., Peroxiredoxins (PRDX), a family of antioxidant enzymes, can catalyze the reduction of various intracellular peroxides, thereby protecting cells from the damage caused by ROS. Mammalian cells contain six PRDX subtypes, of which PRDX6 is the only 1-Cys subclass member, using GSH as the electron donor in the family and with dual enzyme-like activities of glutathione peroxidase and phospholipase A2. effectively remove ROS in cells to maintain the normal state of the body. PRDX6 levels increase in cervical cancer, colorectal cancer, and lung cancer.,, Therefore, we speculate that PRDX6 may be associated with the occurrence and progression of gastric cancer.
In conclusion, PRDX6 knockout BGC-823 cell line was successfully constructed in this study.PRDX6 may be involved in inhibiting the proliferation, migration and invasion of gastric cancer BGC-823 cells by regulating the expression of MMP2 and MMP9. The results are expected to provide a new theoretical basis for the further study of the prevention and treatment of gastric cancer.
| > Subjects and Methods|| |
Research objects, main materials, and reagents
Sixty postoperative paraffin-embedded specimens of primary gastric cancer and 20 normal gastric tissues were collected in the Department of General Surgery, Affiliated Hospital of Beihua University from January 2017 to December 2019, from 38 males and 22 females with an average age of 58.6 years (40–78 years). All the cases were diagnosed as primary gastric cancer by pathological examination and not treated with radiotherapy, chemotherapy, or biological immunotherapy.
Human gastric cancer cell line BGC-823 (Experimental Center, College of Pharmacy, Beihua University, Jilin, China); Roswell Park Memorial Institute (RPMI)-1640 (Gibco Company, USA); newborn bovine serum (Gibico Company, USA); cell counting kit-8 (CCK8) Kit (Sigma Company, USA); trypsin digestive solution (Biyuntian Biotechnology Company, Shanghai, China); lipofectamine®2000 (Invitrogen Company, USA); radioimmunoprecipitation assay buffer (RIPA) lysis buffer (Biyuntian Biotechnology Company, Shanghai, China); dimethyl sulfoxide (Sigma Company, USA); PRDX6 rabbit polyclonal antibody (Invitrogen Company, USA); PRDX6, proliferating cell nuclear antigen (PCNA), Bax, Bcl-2, Matrix metalloproteinases (MMP2), MMP9, PI3K, phospho (p-Akt), and phosphorylated-mammalian target of rapamycin (p-mTOR) monoclonal antibodies (Abcam Company, UK); Transwell chamber (Corning-Costa, USA); and plasmid PIRES (Shenggong Bioengineering Co., Ltd., Shanghai, China).
Immunohistochemical staining and result determination
Gastric cancer and normal gastric tissues were paraffin-embedded to prepare 5-μm slices, and experimental procedures were performed according to the instructions of the immunochemical kit. Determination of results was as follows: PRDX6: No staining was scored as 0, light yellow staining as (1) brown yellow staining as (2) and brown as (3) Grading of positive cell percentage: no positive cell was scored as 0, number of positive cells ≤10% as (1) 11%–50% as (2) 51%–75% as (3) and >75% as (4) Specimens with a result ≥3 points derived from multiplying the scores of the above two items were classified as positive expression cases, and those with a result <3 points were classified as negative expression cases, in which 0–2 points were determined as (−), 3–4 points as (+), 6–8 points as (++), and 9–12 points as (+++).
Cell culture and lentivirus transfection
BGC-823 cells were seeded in RPMI-1640 medium containing 10% fetal bovine serum and penicillin-streptomycin (double antibody, 100 U·mL‒1 penicillin and 100 mg·L‒1 streptomycin), and cultured in an incubator under 5% CO2, 37°C, and 95% saturated humidity. The next day, the culture medium was changed and the culture was continued. The cells were subcultured when the cell coverage rate reached 90%.
Noncoding sh ribonucleic acid (shRNAs) were synthesized by Shanghai Biotechnology Company (China). shRNA-F: 5'-CCTGGAGCAAGGATATCAATGCTTA-3'; shRNA-R: 5'-CCTCGAGAATAGCTATAACGGGTTA-3'. Nucleic acid was cloned into pLKO.1 vector. After transfection into HEK293 cells for 48 h, the virus was released into the medium. The lentivirus was harvested and filtered through a 0.45-μm filter for transfecting gastric cancer BGC-823 cells. The cells were divided into blank control (WT), pLKO.1 vector (vector), and PRDX6 knockout (Short hairpin [sh]-PRDX6) groups.
Quantitative real-time polymerase chain reaction
Total ribonucleic acid (RNA) was extracted using the Trizol method, and absorbance (A) values at 260 and 280 nm were measured with a UV spectrophotometer for calculating the concentration and purity of RNA. RNA was reverse-transcribed into cDNA using the Takara reverse transcription kit, according to the manufacturer's instructions, and quantitative real-time polymerase chain reaction (qRT-PCR) was performed with the SYBR premix exaq. Primer 5.0 was used to design the primers (sense PRDX6-F: 5'-TACGGGCCTCCAG-3'; antisense PRDX6-R: 5'-GCCAAGCTCTGTG-3'; β-actin-F: 5'-GATGAGATTGGCATGGCTTT-3'; β-actin-R: 5'-CACCTTCACCGTTCCAGTTT-3'). The reaction conditions of qRT-PCR: 95°C for 10 min, 95°C for 15 s, 60°C for 30 s and 72°C for 30 s, with 40 cycles. Fluorescence signal acquisition was set at 72°C in the extension step. The melting curve was drawn, for which the settings were 60°C for 30 s, 95°C for 45 s, and 20°C for 10 min. Samples in each group were detected three times, Ct values were recorded, and the 2‒ΔΔCT method was used for analysis.
Detection of proteins expression using western blotting
Cells collected from each group were digested with 0.25% trypsin, the digested cell suspension was centrifuged (1000 g × 5 min), Cells were washed two times with precooled phosphate buffer saline(PBS), and dissolved in RIPA buffer containing phenylmethylsulfonyl fluoride and protease inhibitors. The BCA method was used to determine the protein concentration. A total of 10 μg of protein samples from each group was mixed with five times volume of the protein loading buffer and boiled at 100°C for 5 min for denaturing the proteins. The proteins were transferred onto polyvinylidene fluoride membranes on sodium dodecyl sulfate–polyacrylamide gel electrophoresis, and the membranes were blocked at room temperature withTBS+Tween (TBST) solution containing 5% skimmed milk powder for 2 h. The membranes were incubated with the primary antibody diluents (1:2000) at 4°C overnight, and then washed three times with TBST, 10 min each time. The membranes were incubated with the secondary antibody at 37°C for 1 h, and then washed three times with TBST, 10 min each. After adding the Electro-Chemi-Luminescence (ECL) chemiluminescence solution, the images were developed and photographed. β-actin protein was used as the internal reference, and the optical density of protein bands was measured using the ImageJ software. The final results were expressed as the ratio of the target band and internal reference β-actin.
Detection of cell proliferation with the cell counting kit-8 method
The proliferation activity of BGC-823 cells was detected using the CCK-8 kit. A cell suspension of BGC-823 cells in logarithmic phase was prepared and the cells were seeded in 96-well plates at a density of 1 × 103/well, in which triplicate wells were set in each group. The cells were cultured for 1, 2, 3, 4, and 5 days, CCK-8 (10 μL/well) was added to each well, and the cells were cultured for 3 h. Then, the absorbance (A) value of cells in each group at 450 nm was detected with a microplate reader. The experiment was repeated three times. The value of A represented the proliferation activity of BGC-823 cells, and the cell growth curve was drawn with time as abscissa and A value as ordinate.
BGC-823 cells from the three groups were seeded in 6-well plates at a density of 10 × 106/well and incubated overnight. Then, the cells were digested with 0.25% trypsin, the cell suspension was mixed with a pipette tip, and 250 μL of cells were placed in labeled flow cytometry tubes, in which the blank group was used to adjust the voltage. The cells were diluted and resuspended with ddH2O, and 1 μL fluorescein isothiocyanate annexin V was added into the flow cytometry tubes. Cells were vortexed and incubated at 23°C for 10 min. Then, 1 μL PI solution was added in another single-staining group and experimental group, followed by resuspension with 100 μL binding buffer. The cells were detected on the computer, and data were analyzed and plotted with FlowJo V10.
Cell scratch and transwell invasion test
Scratch test: BGC-823 cells from the three groups were seeded in 6-well plates at a density of 10 × 105/well. The cells were cultured for 24 h, then the supernatant was discarded when the fusion rate of the cells reached 80%–90%, and a wound was made by scratching the monolayer of confluent cells with the tip of a sterile 200-μL pipette, where each well was scratched the same. The wells were washed with PBS three times, and serum-free starvation medium was added to each well. The cells were cultured in a 5% CO2 incubator at 37°C for 24 h, and the experiment was repeated three times independently.
Cell invasion test: BGC-823 cells from the three groups were suspended at a density of 10 × 105/well, and 100 μL of cell suspension was added into the upper chamber. The lower chamber contained a culture medium with 600 μL of 20% FBS. After being cultured for 24 h in an incubator, the upper chamber was taken out and the liquid in the lower chamber was discarded. The cells were stained with Swiss Giemsa staining solution for 90 s and washed with PBS. Cells in five random visual fields were photographed and counted under an optical microscope. All tests were repeated three times.
The SPSS17.0 software (SPSS Inc, USA) was used for statistical analysis. PRDX6 Messenger (mRNA) expression level, cell proliferation activity, wound healing rate, and protein expression level in each group had normal distribution and expressed as mean ± s. The positive rate between the two groups was compared using the t-test. P > 0.05 indicated no significant difference in statistics; P < 0.05 (*) indicated significant difference and P < 0.01 (**) extremely significant difference.
| > Results|| |
Increased expression of peroxiredoxins 6 in gastric cancer tissue
The expression levels and cellular distribution of PRDX6 in the 60 specimens of human gastric cancer and 20 normal gastric tissues were examined by immunohistochemical staining. The results showed that predominantly positive immunostaining was localized in the cytoplasm, with a small amount being present on the cell membrane and nucleus. The positive rate of PRDX6 protein was 65.0% in gastric cancer tissue and 15.0% in normal gastric tissue, suggesting that PRDX6 protein was significantly upregulated in gastric cancer tissue (P < 0.01) [Figure 1]a, [Figure 1]b and [Table 1]. Western blotting was used to detect the expression of PRDX6 in gastric cancer tissue and normal gastric tissue. Compared with that in normal gastric tissue, the expression of PRDX6 in gastric cancer tissue was significantly higher (P < 0.05) [Figure 1]c.
|Figure 1: Immunohistochemical staining of peroxiredoxins 6 in the gastric cancer tissue (a) and normal gastric tissue (b) (×200) and Western blot detection (c). Lane1: gastric cancer tissue, Lane2: normal gastric tissue|
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|Table 1: Comparison of PRDX6 expression between gastric cancer and normal gastric tissues|
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Expressions of peroxiredoxins 6 messenger ribonucleic acid and protein in BGC-823 cells after transfection
Lentivirus-mediated PRDX6 interference was used to knockdown the expression of PRDX6. qRT-PCR results showed that mRNA expression in BGC-823 cells in the sh-PRDX6 group was significantly lower than that in WT and vector groups (P < 0.05) [Figure 2]a. Western blot analysis demonstrated that the expression of PRDX6 protein in BGC-823 cells in the sh-PRDX6 group was significantly lower than that in WT and vector groups (P < 0.05) [Figure 2]b.
|Figure 2: Peroxiredoxins 6 messenger ribonucleic acid (a) and protein (b) expression of BGC-823 cells transfected with lentivirus. Lane1: WT group; Lane2: Vector group; Lane3: Short hairpin-peroxiredoxins 6 group|
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Effect of peroxiredoxins 6 on the proliferation of BGC-823 cells in vitro
Results from the CCK8 assay revealed that the viability of BGC-823 cells in the sh-PRDX6 group was significantly lower than that in WT and vector groups on the 3rd, 4th, and 5th day after lentivirus infection (P < 0.05) [Figure 3]a. Western blot analysis showed that the expression of PCNA, a proliferation-associated protein, in the sh-PRDX6 group was significantly lower (P < 0.05) [Figure 3]b, suggesting that PRDX6 promotes the growth of gastric cancer cells by upregulating PCNA.
|Figure 3: Proliferation activities of BGC-823 cells in various groups. (a) Proliferation of BGC-823 detected by cell counting kit-8; (b) Expression of proliferating cell nuclear antigen protein detected by Western blot. Lane1: WT group; Lane2: Vector group; Lane3: Short hairpin-peroxiredoxins 6 group|
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Effect of peroxiredoxins 6 on BGC-823 cell apoptosis
As shown in [Figure 4]a, [Figure 4]b, [Figure 4]c, the apoptotic rate of BGC-823 cells measured by flow cytometry in the sh-PRDX6 group (21.61% ± 0.18%) was significantly increased compared with that in WT (5.85% ± 0.11%) and vector (5.79% ± 0.09%) (P < 0.05) groups, indicating that PRDX6 silencing could significantly increase BGC-823 cell apoptosis. Western blot analysis revealed that the expression level of Bax in BGC-823 cells was significantly increased and that of Bcl-2 was significantly decreased in the sh-PRDX6 group compared with that in WT and vector groups (P < 0.05) [Figure 4]d.
|Figure 4: Effects of peroxiredoxins 6 on the apoptosis and apoptosis-related protein expressions (d) in BGC-823 cells. (a) WT group; (b) Vector group; (c) sh-PRDX6 group. Lane1: WT group; Lane2: Vector group; Lane3: Short hairpin-peroxiredoxins 6 group|
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Effect of peroxiredoxins 6 transfection on the migration and invasion of BGC-823 cells in vitro
Effects of PRDX6 on the migration and invasion of BGC-823 cells were tested using the wound healing and transwell assays. As shown in [Figure 5]a and [Figure 5]b, the wound healing rate of BGC-823 cells in the sh-PRDX6 group (23.4% ± 0.2%) was significantly decreased compared with that in WT (52.6% ± 0.4%) or vector (50.8% ± 0.3%) groups at 24 h after PRDX6 transfection (P < 0.05). Results from the transwell assay revealed that the number of migrating BGC-823 cells in the sh-PRDX6 group (135.6 ± 12.3) was significantly reduced compared with that in WT (81.3 ± 9.5) or vector (84.8 ± 10.2) (P < 0.05) groups. Western blot analysis showed that the expression of MMP2 and MMP9 in the sh-PRDX6 group was significantly higher than that in WT and vector groups (P < 0.05) [Figure 5]c. These findings indicate that PRDX6 knockdown could significantly decrease the migration and invasion abilities of BGC-823 cells.
|Figure 5: Effects of peroxiredoxins 6 on the migration and invasion of BGC-823 cells in vitro. (a) Effect of peroxiredoxins 6 on the migration rate of BGC-823 cells in vitro; (b) Effect of peroxiredoxins 6 on the invasion of BGC-823 cells in vitro; (c) Expressions of matrix metalloproteinases 2 and matrix metalloproteinases 9 proteins by Western blot. Lane1: WT group; Lane2: Vector group; Lane3: Short hairpin-peroxiredoxins 6 group|
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Effects of peroxiredoxins 6 on the PI3K/Akt/mammalian target of rapamycin signaling pathway in BGC-823 cells
Western blotting was used to detect the expression of related proteins before and after PRDX6 knockdown [Figure 6]. Compared with those in WT and vector groups, the expression levels of PI3K, p-Akt, and p-mTOR proteins in the sh-PRDX6 group were significantly decreased (P < 0.05), whereas those of unphosphorylated Akt and mTOR were not significantly different, indicating that PRDX6 participates in the regulation of the PI3K/Akt/mTOR signaling pathway in BGC-823 cells.
|Figure 6: Expressions of PI3K/Akt/mTOR signaling pathway-related proteins detected by Western blot. Lane1: WT group; Lane2: Vector group; Lane3: sh-PRDX6 group|
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| > Discussion|| |
Cancer is usually caused by a persistent imbalance of the redox state in cells, and cancer cells often are in a higher redox state than normal cells. PRDX is a kind of selenium-independent peroxidase family protein found in many organisms. Studies on the expression and function of PRDX in tumors have gradually become a hotspot in recent years. PRDX has been found to be upregulated in many cancers and is involved in almost all processes from the growth and metastasis of tumors to the regulation of their response to treatment. Among PRDX family members, PRDX6 is a bifunctional protein with dual activities of GSH peroxidase and calcium-independent phospholipase A2. PRDX6 can reportedly not only inhibit the carcinogenesis of hepatoma cells but also promote tumor necrosis factor-A-induced apoptosis in cancer cells; PRDX6 is highly expressed in lung cancer tissues, and PRDX6 overexpression can promote the proliferation, invasion, and migration of lung cancer cells; PRDX6 is highly expressed in melanoma cell lines and promotes the growth of melanoma cells by triggering the arachidonic acid-dependent lipid signaling pathway. These results suggest that PRDX6 plays an important role in the proliferation, apoptosis, and migration of gastric cancer cells. Thus, our findings may provide a new target for the diagnosis, treatment, and prognosis of gastric cancer.
It has been PRDX6 is highly expressed in many tumors, and its overexpression increases the resistance of tumor cells to chemoradiotherapeutic agents; knockdown of PRDX6 can increase the chemosensitivity of tumor cells by inducing apoptosis. In this study, immunohistochemistry and western blotting were used to detect the expression of PRDX6 in gastric cancer and normal gastric tissues, respectively. The findings revealed that the expression of PRDX6 in gastric cancer tissue was significantly increased, suggesting that PRDX6 may be involved in the carcinogenesis of normal gastric tissue. To further study the biological function of PRDX6 in gastric cancer cells, we constructed a PRDX6 knockdown stable cell line using lentivirus for subsequent experiments. The CCK8 method was used to detect the effect of PRDX6 on the proliferation of gastric cancer cells. The results showed that the downregulation of PRDX6 could significantly inhibit the proliferation of gastric cancer cells. Moreover, PRDX6 protein could affect the proliferation of gastric cancer cells by affecting PCNA protein.
Invasion and metastasis, important features of malignant tumor, are complex processes, involving a variety of mechanisms. MMPs, a family of zinc-dependent proteolytic enzymes, are involved in the invasion, metastasis, and angiogenesis of tumors. MMP2 and MMP9, the most widely studied MMPs, play an important role in the invasion and metastasis of tumors. Many studies have confirmed that MMP2 can promote the migration and invasion of cancer cells by helping tumor cells escape immune surveillance. MMP9 can promote tumor cells to secrete transforming growth factor-β to promote the proliferation, migration, and invasion of tumor cells. The results showed that compared with the control group, in the sh-PRDX6 group, BGC-823 cell migration and invasion were significantly decreased, and the expression levels of MMP2 and MMP9 were significantly decreased. Combined with previous reports, we hypothesize that PRDX6 may inhibit the occurrence and progression of gastric cancer by affecting the expression and secretion of MMP2 and MMP9.
To some extent, apoptosis is regulated by antiapoptotic Bcl-2 and proapoptotic Bax. Bcl-2 can lead to structural and functional disorders of the mitochondria by inhibiting Bax to block the release of caspase, and then inhibit apoptosis. The results showed that the knockdown of PRDX6 protein expression in BGC-823 cells resulted in the downregulation of Bcl-2 expression and the upregulation of Bax expression to promote BGC-823 cell apoptosis, suggesting that PRDX6 silencing induces apoptosis through the mitochondrial pathway.
PI3K/Akt/mTOR signaling pathway is an important pathway for regulating cell cycle, proliferation, apoptosis, differentiation, and protein transport and has always been a research hotspot in the study of the biological behavior of tumor cells., Many studies have shown that the PI3K/Akt/mTOR signaling pathway can accelerate the activity of tumor cells and regulate the receptors to promote the proliferation and metastasis of tumor cells. In this study, the expression of PRDX6 could significantly decrease the expression levels of PI3K, p-Akt, and p-mTOR in BGC-823 cells, suggesting that PRDX6 regulates the biological behavior of gastric cancer cells through the PI3K/Akt/mTOR signaling pathway.
Furthermore, our findings revealed that PRDX6 gene knockdown mediated by lentivirus could inhibit the proliferation, migration, and invasion of BGC-823 cells and promote apoptosis. The mechanism may be related to the regulation of PI3K/Akt/mTOR signaling protein expression. In conclusion, PRDX6 may be a potential target for the clinical diagnosis and treatment of gastric cancer. However, the other functions of PRDX6 and its molecular mechanism need to be further explored.
We acknowledge the doctors of Department of General Surgery (Affiliated Hospital of Beihua University, Jilin, China) for sample collection and processing. This work was supported by National Natural Science Foundation of China under grant: No. 31900918; Science and Technology Bureau of Jilin Province grants: 20190303047SF and 20170520033JH; It also supported by Jilin Department of Health grant: 2019Q020and Graduate Innovation Program of Beihua University grant: 2021019 and 2021021 and College students' Innovation Project grant 202110201172, 202110201067 and 202110201090.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| > References|| |
Jemal A, Siegel R, Xu J, Ward E. Cancer statistics, 2010. CA Cancer J Clin 2010;60:277-300.
Shah R, Khaitan PG, Pandita TK, Rafiq A, Abrol D, Suri J, et al.
Gastric cancer in Jammu and Kashmir, India: A review of genetic perspectives. J Cancer Res Ther 2020. doi: 10.4103/jcrt.JCRT_12_19.
Nakamura M, Nakamori M, Ojima T, Katsuda M, Yamaue H. Conversion surgery for advancedgastric cancer with peritoneal metastasis. J Clin Oncol 2020;38 Suppl 4:450.
Sakin A, Samanci NS, Secmeler S, Arici S, Can O, Yasar N, et al.
The effect of body mass index on location of recurrence and survival in early-stage colorectal cancer. J Cancer Res Ther 2020;16:S176-82.
García-Magallanes N, Beltran-Ontiveros SA, Leal-León EA, Luque-Ortega F, Romero-Quintana JG, Osuna-Ramirez I, et al.
Underexpression of circulating miR-145-5p and miR-133a-3p are associated with breast cancer and immunohistochemical markers. J Cancer Res Ther 2020;16:1223-8.
Ighodaro OM, Akinloye OA. First line defence antioxidants-Superoxide Dismutase (SOD), Catalase (CAT) and Glutathione Peroxidase (GPX): Their fundamental role in the entire antioxidant defence grid. Alexandria Journal of Medicine 2018;54:287-93. DOI: 10.1016/j.ajme.2017.09.001.
Toppo S, Flohé L, Ursini F, Vanin S, Maiorino M. Catalytic mechanisms and specificities of glutathione peroxidases: Variations of a basic scheme. Biochim Biophys Acta 2009;1790:1486-500.
Kim H, Seo S, Kim K, Park YH, An M, Baik H, et al.
Prognostic significance of Human epidermal growth factor receptor-2 expression in patients with resectable gastric adenocarcinoma. World J Surg Oncol 2019;17:122.
Ruggieri V, Russi S, Zoppoli P, La Rocca F, Angrisano T, Falco G, et al.
The role of MicroRNAs in the regulation of gastric cancer stem cells: A meta-analysis of the current status. J Clin Med 2019;8:639.
Rhee SG, Woo HA, Kil IS, Bae SH. Peroxiredoxin functions as a peroxidase and a regulator and sensor of local peroxides. J Biol Chem 2012;287:4403-10.
Fisher AB. Peroxiredoxin 6: A bifunctional enzyme with glutathione peroxidase and phospholipase A2 activities. Antioxid Redox Signal 2011;15:831-44.
Hu X, Lu E, Pan C, Xu Y, Zhu X. Overexpression and biological function of PRDX6 in human cervical cancer. J Cancer 2020;11:2390-400.
Huang WS, Huang CY, Hsieh MC, Kuo YH, Tung SY, Shen CH, et al.
Expression of PRDX6 correlates with migration and invasiveness of colorectal cancer cells. Cell Physiol Biochem 2018;51:2616-30.
Jo M, Yun HM, Park KR, Hee Park M, Myoung Kim T, Ho Pak J, et al.
Lung tumor growth-promoting function of peroxiredoxin 6. Free Radic Biol Med 2013;61:453-63.
Kim SY, Chun E, Lee KY. Phospholipase A (2) of peroxiredoxin 6 has a critical role in tumor necrosis factor-induced apoptosis. Cell Death Differ 2011;18:1573-83.
Yun HM, Park KR, Lee HP, Lee DH, Jo , Shin DH, et al.
PRDX6 promotes lung tumor progression via its GPx and iPLA2 activities. Free Radic Biol Med 2014;69:367-76.
Schmitt A, Schmitz W, Hufnagel A, Schartl M, Meierjohann S. Peroxiredoxin 6 triggers melanoma cell growth by increasing arachidonic acid-dependent lipid signalling. Biochem J 2015;471:267-79.
Yun HM, Park KR, Park MH, Kim DH, Jo MR, Kim JY, et al.
PRDX6 promotes tumor development via the JAK2/STAT3 pathway in a urethane-induced lung tumor model. Free Radic Biol Med 2015;80:136-44.
Aparna M, Rao L, Kunhikatta V, Radhakrishnan R. The role of MMP-2 and MMP-9 as prognostic markers in the early stages of tongue squamous cell carcinoma. J Oral Pathol Med 2015;44:345-52.
Stellas D, Hamidieh AE, Patsavoudi E. Monoclonal antibody 4c5 prevents activation of mmp2 and mmp9 by disrupting their interaction with extracellular hsp90 and inhibits formation of metastatic breast cancer cell deposits. BMC Cell Biol 2010;11:51.
Zhao SC, Wu QW, Cheng YF, Jia YF, Chen J, Yan CL. Protective effect and mechanism of butylphthalide on MPP + - induced apoptosis of SH-SY5Y cells[J]. Chinese Journal of Gerontology, 2017;(37): 316-8.(in Chinese)
Meng Y, Hu J, Chen Y, Yu T, Hu L. Silencing MARCH1 suppresses proliferation, migration and invasion of ovarian cancer SKOV3 cells via downregulation of NF-κB and Wnt/β-catenin pathways. Oncol Rep 2016;36:2463-70.
Xie L, Dai H, Li M, Yang W, Yu G, Wang X, et al.
MARCH1 encourages tumour progression of hepatocellular carcinoma via regulation of PI3K-AKT-β-catenin pathways. J Cell Mol Med 2019;23:3386-401.
Zhou BP, Liao Y, Xia W, Spohn B, Lee MH, Hung MC. Cytoplasmic localization of p21Cip1/WAF1 by Akt-induced phosphorylation in HER-2/neu-overexpressing cells. Nat Cell Biol 2001;3:245-52.
[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6]