|Year : 2016 | Volume
| Issue : 8 | Page : 248-251
Leucine-rich repeats and immunoglobulin-like domains protein 1 and fascin actin-bundling protein 1 expression in nonsmall cell lung cancer
Juanjuan Zhang, Xiaoyan Wang, Yijie Zhang, Jingcan Wu, Nannan Zhou
Department of Respiratory, Henan University Huaihe Hospital, Kaifeng, Henan, China
|Date of Web Publication||22-Feb-2017|
Department of Respiratory, Henan University Huaihe Hospital, Kaifeng 475000, Henan
Source of Support: None, Conflict of Interest: None
Objective: To assess the clinical significance of leucine-rich repeats and immunoglobulin-like domains protein 1 (LRIG1) and fascin actin-bundling protein 1 (Fascin-1) expression in nonsmall cell lung cancer (NSCLC).
Materials and Methods: Six-one NSCLC patients were included in this study. The expression of LRIG1 and Fascin-1 was assayed in the tumor tissue and relative normal lung tissue of the 61 NSCLC patients by immunohistochemistry. The relationship between LRIG1, Fascin-1 expression pattern and lung cancer patients' clinical pathology characteristics was evaluated.
Results: The positive expression rate of Fascin-1 in cancer tissue and normal tissue was 70.5% (43/61) and 13.1% (8/61), respectively, which indicated cancer tissue much higher than normal tissue (P < 0.05); for LRIG1, the positive expression rate was 54.1% (33/61) and 82.0% (50/61) for tumor tissue and normal tissue with statistical difference (P < 0.05); Fascin-1-positive expression was associated with tumor diameter (P < 0.05) and mediastinal lymph node metastasis (P < 0.05). Moreover, LRIG1-positive expression was correlated with pathology type (P < 0.05), clinical stage (P < 0.05), and mediastinal lymph node metastasis (P < 0.05).
Conclusion: LRIG1 and Fascin-1 were differently expressed in cancer and normal lung tissue in patients with NSCLC, which could be a biomarker for mediastinal lymph node metastasis in NSCLC patients.
Keywords: Clinical pathology, fascin actin-bundling protein 1, immunohistochemistry, leucine-rich repeats and immunoglobulin-like domains protein 1, nonsmall cell lung cancer
|How to cite this article:|
Zhang J, Wang X, Zhang Y, Wu J, Zhou N. Leucine-rich repeats and immunoglobulin-like domains protein 1 and fascin actin-bundling protein 1 expression in nonsmall cell lung cancer. J Can Res Ther 2016;12:248-51
|How to cite this URL:|
Zhang J, Wang X, Zhang Y, Wu J, Zhou N. Leucine-rich repeats and immunoglobulin-like domains protein 1 and fascin actin-bundling protein 1 expression in nonsmall cell lung cancer. J Can Res Ther [serial online] 2016 [cited 2017 Mar 26];12:248-51. Available from: http://www.cancerjournal.net/text.asp?2016/12/8/248/200749
Juanjuan Zhang and Xiaoyan Wang contributed equally to this work.
| > Introduction|| |
Lung cancer is one of the leading causes of cancer-related death worldwide. It was reported that 220,000 lung cancer patients were diagnosed in the year 2013 and about 150,000 dead of lung cancer the same year. In the year 2008, the Ministry of Health of China reported the survey on causes of the residents' death. According to the report, lung cancer had become one of the highest morbidities and mortalities of malignant tumors in our country. However, the etiology of lung cancer is still not fully understood. Therefore, to explore the molecular mechanism of lung cancer development, invasion, and metastasis and to provide new ideas and new targets for further treatment of lung cancer have become a hot research topic.
Leucine-rich repeats and immunoglobulin-like domains protein 1 (LRIG1) is a protein that in humans is encoded by the LRIG1 gene, which has been reported to interact with receptor tyrosine kinases of the EGFR-family. Moreover, several studies have found that LRIG1 expression was associated with lung cancer prognosis., Fascin actin-bundling protein 1 (Fascin-1) is an actin-binding protein, and it was found that the expression of Fascin-1 was increased in multiple human epithelial tumors. In the present study, we discussed the clinical significance of LRIG1 and Fascin-1 expression in nonsmall cell lung cancer (NSCLC) and their association with the clinical characteristics.
| > Materials and Methods|| |
Sixty-one patients with pathology confirmed NSCLC were included in this study. The expression of LRIG1 and Fascin-1 was assayed in the tumor tissue and relative normal lung tissue of the 61 NSCLC patients by immunohistochemistry. The mean age of the included 61 patients was 60.12 ± 11.4 years, with 40 males and 21 females. There were 46 cases with Stage I/II and 15 cases with Stage III. For the pathology type, there were 30 cases of adenocarcinoma, 21 cases of squamous cell carcinoma, 2 cases of big cell carcinoma, and 2 cases of adenosquamous carcinoma.
Reagents and instruments
Ultra-Thin Semiautomatic Microtome (from Leica, Germany), microscope (from Olympus Corporation, Japan), electrothermostat (from Shanghai Jianheng Instrument Co., Ltd., China), microwave oven, low-speed centrifuge (Beckman Coulter, Inc., USA), mouse anti-human Fascin-1 monoclonal antibody (SPM133; Santa Cruz, USA), mouse anti-human LRIG1 monoclonal antibody (Ab36704; Abcam, UK), ready-to-use immunohistochemical assay kit, (from Sangon Biotech Co., Ltd., Shanghai, China), DAB display kit (from Solar Biotech, Beijing, China), second antibody (goat anti-rabbit IgG; Santa Cruz, USA) were used.
Immunohistochemical was detected according to the following steps. Paraffin slices were sliced and hydrated. Antigen recovery was made after washing with distilled water. Animal nonimmune serum was added after phosphate-buffered saline (PBS) irrigation; after incubation, it was poured out. The first antibody was added drop-wise and incubated at room temperature for 60 min. The negative control was then replaced by PBS. After washing with PBS, the biotin-labeled as secondary antibody was added and incubated at room temperature for 10 min. After washing with PBS, Streptomyces anti-biotin streptavidin-peroxidase was added and incubated at room temperature for 30 min. It was washed with PBS and developed with DAB agent. Finally, pieces were sealed with a neutral gum.
Positive result determination
Fascin-1 protein is mainly localized in the cytoplasm or membrane, showing a focal or diffuse yellow granular shape; LRIG1 is mainly expressed in the nucleus with yellow staining, showing a diffuse distribution. Under the microscope, ten fields are in each slice, counting the total number of cells and the number of positive cells in each field. The criteria for the percentage of positive cells are as follows: 0%–5%, negative expression; 6%–25%, weakly positive; 26%–75%, moderately positive; and above 76%, strongly positive. The positive result criteria include weakly positive, moderately positive, and strongly positive.
All the statistical analyses were done by SPSS 13.0 statistical software (SPSS Inc., Chicago, IL, USA). The comparison of sample rate was processed by χ2 test; P < 0.05 was deemed as statistical significance.
| > Results|| |
Leucine-rich repeats and immunoglobulin-like domains protein 1 and fascin actin-bundling protein 1 expression
Immunohistochemical staining showed that Fascin-1 protein was mainly localized in the cytoplasm or membrane of the cells with focal or diffused yellow granule. The positive expression rate of Fascin-1 in cancer tissue and normal tissue was 70.5% (43/61) and 13.1% (8/61), respectively, which indicated cancer tissue was much higher than normal tissue (P < 0.05); For LRIG1, the positive expression rate was 54.1% (33/61) and 82.0% (50/61) for tumor tissue and normal tissue, with statistical difference (P < 0.05) [Table 1].
|Table 1: The positive rate of fascin actin.-bundling protein 1 and fascin actin.-bundling protein 1 in cancer and normal tissue|
Click here to view
Correlation between leucine-rich repeats and immunoglobulin-like domains protein 1, fascin actin-bundling protein 1 expression, and clinical characteristics
Fascin-1-positive expression was associated with tumor diameter (P< 0.05) and mediastinal lymph node metastasis (P< 0.05). Moreover, LRIG1-positive expression was correlated with pathology type (P< 0.05), clinical stage (P< 0.05), and mediastinal lymph node metastasis (P< 0.05) [Table 2].
|Table 2: The association between leucine-rich repeats and immunoglobulin-like domains protein 1, fascin actin-bundling protein 1 expression and clinical characteristics|
Click here to view
| > Discussion|| |
With the recent progress of molecular biology research, it is proven that the expression of more and more protein factors is related to the clinicopathological features and prognosis of patients, such as epidermal growth and vascular endothelial growth factor. In recent years, many studies have found that Fascin-1 protein is highly expressed in many kinds of solid tumors, such as lung, intestinal, liver, and prostatic cancer; this protein is certainly related to the clinicopathological features of patients.,, Gene-encoding fascin protein is localized in the human chromosome 7p22. Fascin-1 protein is a member of the cytoskeletal protein family. This protein can strengthen the actin cytoskeleton to develop resistance to the cell membrane elongation because of cell protrusion movement during cell migration, thereby playing an important role in cell migration. Recent studies show that Fascin-1 is highly expressed in various tumor tissues. Kvarnbrink et al. studied the expression of Fascin-1 in esophageal cancer using the immunohistochemical method. The expression of Fascin-1 in tumor tissues is much higher than its expression in normal esophageal epithelium. The researchers believe that the expression of Fascin-1 can be used to help judge the transformation of normal esophageal epithelium tissues to tumor tissues. Rui et al. detected the expression of Fascin-1 protein in 84 cases of lung cancer tissues and adjacent tissues. The result shows that the positive expression of Fascin-1 in tumor tissues is higher than that in adjacent tissues, and the positive expression is related to the age, N stage, and clinical staging of patients. In this paper, it is found that Fascin-1 protein is mainly localized in the cytoplasm or membrane and shows a focal or diffuse yellow and granular shape. The positive expression rates in tumor tissues and adjacent tissues are 70.5% (43/61) and 13.1% (8/61), respectively. The positive rate in tumor tissues is significantly higher than that in adjacent tissues. This finding is consistent with the results of the study conducted by Rui et al. Therefore, Fascin-1 may be involved in the occurrence and development of large cells in lung cancer, lymph node metastasis, and other pathological processes.
LRIG is a newly discovered anti-oncogene, which widely exists in mammals. Its encoded product is a transmembrane glycoprotein. Many LRIG has been found in its extracellular segment. At present, the study on LRIG is more thorough than before, and studies show that it is more closely related to various malignant tumors. Fei et al. transfected LRIG1 gene into glioma cell line H4 by means of liposome-mediated gene transfection technology. The study shows that mRNA and protein of the transfected LRIG1 gene are significantly increased. The proliferation of glioma cells after transfection is obviously inhibited; apoptosis is significantly increased. Compared with the nontransfected LRIG1 gene, the difference has statistical significance. The relative percentage of H4 cells transfected with LRIG1 through the artificial recombinant basement membrane is significantly lower than the percentage of cells through the membrane of the nontreatment and mock-vehicle groups. The difference has statistical significance.
However, a few reports have been presented on the expression of LRIG1 in large lung tumors and its clinical significance. We detect the expression of LRIG1 protein in lung tumor tissues and adjacent tissues using the immune combination. The expression of LRIG1 in tumor tissues is significantly lower than in adjacent tissues, and the clinical staging of the patients with low expression in tumor tissues is done late (with higher lymph node metastasis). Results indicate that LRIG1 may be involved in the occurrence of lung cancer and lymph node metastasis.
Therefore, it is believed that LRIG1 and Fascin-1 are expressed differently in lung cancer tissues and normal lung tissues adjacent to the tumor. They are also related to the occurrence and development of large cells in lung cancer, as well as lymph node metastasis. Further study on the above-mentioned proteins and their expression regulation mechanism will help develop a targeted therapy for large lung tumors.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| > References|| |
Adams JC. Fascin-1 as a biomarker and prospective therapeutic target in colorectal cancer. Expert Rev Mol Diagn 2015;15:41-8.
Kou C, Zhou T, Han X, Zhuang H, Qian H. LRIG1, a 3p tumor suppressor, represses EGFR signaling and is a novel epigenetic silenced gene in colorectal cancer. Biochem Biophys Res Commun 2015;464:519-25.
An Y, Zhao Z, Ou P, Wang G. Expression of LRIG1 is associated with good prognosis for human non-small cell lung cancer. Medicine (Baltimore) 2015;94:e2081.
Gur G, Rubin C, Katz M, Amit I, Citri A, Nilsson J, et al.
LRIG1 restricts growth factor signaling by enhancing receptor ubiquitylation and degradation. EMBO J 2004;23:3270-81.
Kvarnbrink S, Karlsson T, Edlund K, Botling J, Lindquist D, Jirström K, et al.
LRIG1 is a prognostic biomarker in non-small cell lung cancer. Acta Oncol 2015;54:1113-9.
Omran OM, Al Sheeha M. Cytoskeletal focal adhesion proteins Fascin-1 and paxillin are predictors of malignant progression and poor prognosis in human breast cancer. J Environ Pathol Toxicol Oncol 2015;34:201-12.
Siegel R, Naishadham D, Jemal A. Cancer statistics, 2013. CA Cancer J Clin 2013;63:11-30.
Zhang FR, Tao LH, Shen ZY, Lv Z, Xu LY, Li EM. Fascin expression in human embryonic, fetal, and normal adult tissue. J Histochem Cytochem 2008;56:193-9.
Zhao W, Gao J, Wu J, Liu QH, Wang ZG, Li HL, et al.
Expression of Fascin-1 on human lung cancer and paracarcinoma tissue and its relation to clinicopathological characteristics in patients with lung cancer. Onco Targets Ther 2015;8:2571-6.
Rui W, Hongzhen Y, Xuan W. Fascin-1 expression in lung cancer and its clinical significance. Acta Univ Med Anhui 2014;25:658-61.
Fei Y, Dongshegn G, Hongquan N. Molecular mechanism of LRIG1 cDN-induced apoptosis in human glioma cell line H4. Chin J Cancer 2004;10:1149-54.
[Table 1], [Table 2]