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Putative role of epithelial membrane protein-1 in the pathogenesis of colorectal carcinoma
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Abstract:
Objective: To elucidate the putative involvement of epithelial membrane protein-1 (EMP1) in the pathogenesis of colorectal carcinoma (CRC). Methods: Tumor ( n =63) and peri-tumor ( n =31) tissue specimens were collected from 63 patients with CRC. EMP1 protein content in these specimens was assessed by immunohistochemistry and Western blotting. The relationship between EMP1 protein content in the tumor tissue and the pathologic grade of the lesions was analyzed. To further evaluate the putative role for EMP1 in the development of CRC, we also performed analysis in vitro . To this end, CRC SW-480 cells were transfected with an EMP1 lentiviral vector pLenti6-EMP1 or a control vector plenti6/V5-DEST. EMP1 mRNA abundance and protein content in transfected cells were determined by quantitative real-time PCR and Western blotting respectively. Effects of EMP1 overexpression on the proliferation, apoptosis and invasion of SW-480 cells were evaluated by methyl thiazolyl tetrazoliun (MTT) assay, flow cytometry (FCM) and transwell invasion assays, respectively. Results: EMP1 protein content was significantly lower in CRC tissue than in peritumor tissues ( P <0.05). The level of EMP1 protein was not correlated with gender, age, and tumor location ( P >0.05) but was positively correlated with lymph node metastasis, clinic stage and histological grade of the lesions ( P <0.05). Compared with SW-480 cells transfected with the control vector, SW-480 cells overexpressing EMP1 had a lower survival fraction (\[60.94±4.04\]% vs \[100.00±0.00\]%， P <0.05), a higher cell apoptosis rate (\[12.10±1.30\]% vs \[3.10±060\]%， P <0.05), a decreased invasive capacity (\[178.00±21.00\] vs \[87.00±12.00\], P <0.05), higher caspase-9 protein content (0.764±0.073 vs 0.231±0.029, P <0.05) and lower VEGFC protein content (0.663±0.065 vs 0.185±0022, P <0.05). Conclusion: EMP1 protein content is significantly lower in the CRC tissue than in the non-CRC tissue. Overexpression of EMP1 in CRC cells in vitro results in a significant inhibition of invasive activity through regulating the expression of caspase-9 and VEGF-C. These findings suggest that EMP1 is involved in the pathogenesis of colorectal carcinoma.
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