| ObjectiveOvarian cancer is the top 10 common cancer in China.It ranks third in incidence rate in malignant tumor of the female reproductive system,while its mortality just behind cervical cancer.The reason is that it’s often asymptomatic before ovarian cancer occurrence and metastasis-Because most of the ovarian cancer patients are diagnosed with terminal stage of cancer, they miss the most effective treatment.Therefore,finding the specific genes associated with ovarian cancer provides new theoretical basis for the diagnosis and treatment of ovarian cancer.For this purpose,a large number of research dedicated to finding specific genes associated with ovarian cancer at home and abroad. TMEM(Transmembrane Protein) as a part of the membrane proteins involve in cell signal transduction, proliferation and differentiation.Therefore it becomes one of research hot spot for cancer related gene.Most of TMEM family are cancer genes, such as TMEM16a, TMEM17a, TMEM17b, and some are opposite,such as TMEM100.The function of TMEM49 in different types of human tumors has been recently reported,but the association of TMEM49 and ovarian cancer is poorly defined. Here,we detect the expression of TMEM49 in normal ovarian tissue and ovarian cancer through immunohistochemical staining and explore the relationship between the expression of TMEM49 in ovarian cancer to figure out its clinical significance. Furthermore,we explore its influence on the biological behavior of ovarian cancer cell by RNA interference to study its relevance to ovarian cancer occurrence, development and metastasis, and reveal the molecular mechanism in the process of cancer and provide a new idea for targeted therapy of ovarian cancer.MethodsImmunohistochemical staining was used to detect the expression of TMEM49 in 30 ovarian cancer specimens and also normal ovarian tissure. Western blot and qRT-PCR were used to detect the level of TMEM49 expression in five different ovarian cancer cell lines, then we selected the high TMEM49-expression cell line to construct TMEM49-shRNA stable transfection cell line. Cell proliferation, cell cycle and apoptosis,cell metastasis and adhesion experiment were conducted to explore the influence of TMEM49 for cell biological behavior of ovarian cancer cell. And then using qRT-PCR and Western blot to analyse the expression of Caspase3, Bad and Bax,which in apoptosis pathway,and MMP2,KLF10 and CXCL12 which in metastasis pathway in different cell line,in order to detect TMEM49 interference for this two pathway in ovarian cancer cell.Results1.TMEM49 expression in ovarian cancer tissues was higher than normal ovarian tissue, and there was a significant difference(P<0.05).2.The expression levels of TMEM49 was related with the FIGO surgical stage and histological grade in ovarian carcinoma (P<0.05). The higher FIGO surgical stage and histological grade,the more expression of TMEM49.3.Down-regulation of TMEM49 through RNA interference inhibited cell proliferation, arrested Gl/S transition and induced cell apoptosis in ovarian cancer cell line. Additionally, down-regulation of TMEM49 in ovarian cancer cell notably repressed cell invasion and adhesion.All of these are significant difference(P<0.05).4. Western blot and PCR revealed the expression of Caspase3, Bad and Bax were increased, MMP2, KLF10 and CXCL12 were reduced when silencing TMEM49 by RNA interference(P<0.05).Conclusion1.TMEM49 was overexpressed in ovarian cancer compared to normal ovarian tissue, and was associated with higher FIGO surgical stage and histological grade of disease. It may because of overexpression of TMEM49 involve in tumor growth of ovarian cancer.2.Knockdown of TMEM49 could reduce cell proliferation,metastasis and invasion,arrest G1/S transition,and induce cell apoptosis,it showed that TMEM49 may play a key role in the growth of ovarian cancer, thus TMEM49 may be a potential therapeutic target in human ovarian cancer.3.Silencing TMEM49 could increase the expression of Caspase3, Bad and Bax, which in apoptosis pathway,and surpress MMP2,KLF10 and CXCL12 which in metastasis pathway.It may indicate that TMEM49 seems to be associated with the apoptosis and metastasis up signaling in growth and progression of ovarian cancer. |
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