Effect Of RNAi Targeting CLIC1 On The Physiological Function Of Lung Adenocarcinoma Cells

Lung cancer is the most life-threatening neoplasia in the world with high incidence and mortality rates. Especially, human lung adenocarcinoma became more and more prevalent in recent years. Lung cancer derived from lung epithelial; the process of neoplasm formation may take 20-30 years. The tumorigenesis features heterogeneity and metastasis, most of patiences were diagnosed and treated until late stages, contributing to the low efficiency of therapy and bad prognosis. Based on the analytic results from differential proteomics, we choose chloride intracellular channel protein 1 (CLIC1) that has over-expression in lung adenocarcinoma for further functional studies, aiming to reveal whether CLIC1 functions as a potential biomarker for prognosis and a targeted protein for anti-cancer treatment of lung adenocarcinoma.AIM:To observe the mRNA level and protein expression level of CLIC1 after transiently transfecting human lung carcinoma cell A549 using siRNA, to detect the effect of RNAi targeting CLIC1 on the proliferation and metastasis of lung carcinoma cell A549. The effect of RNAi targeting CLIC1 on physiological function was investigated.Methods:Real-time quantitative polymerase chain reaction and Western blot methods were used to detect the knock down of CLIC1 mRNA level and protein expression level after transfection for 0,2,48,and 72 h. WST-1 method was used to detect the proliferation of the cells every day in five days after transfection and the cell growth curve was depicted. Cell cycle and apoptosis were determined on cells via flow cytometry after transfecting 48 h. Transwell chamber and scrach healing were used to analyze the migration and invasion of A549 cells.Results:The mRNA level of CLIC1 was knocked down 97% after transfecting 24 h; the protein level of CLIC1 was knocked down over 80% after transfecting 72 h. Compared with two control groups, cell proliferation of the siRNA group was suppressed (P＜0.05); however, cell cycle and primitive apoptosis of the other three groups did not change dramatically after transfecting 48 h. At the same time, the migration and invasion of the siRNA group were suppressed 60%(P<0.01), and-68%(P＜0.05), respectively.Conclusion:Silencing CLIC1 could effectively inhibit the proliferation of human lung carcinoma cells and suppress the metastasis of the tumor. CLIC1 could be a new target for suppressing the proliferation and the metastasis of the tumor.