The influence of silencing Ezrin expression by RNA interference on the biological behaviors of colorectal carcinoma in vitro / 肿瘤

ABSTRACT

Objective: To study the role of membrane-cytoskeleton linker Ezrin in the growth and metastasis of human colorectal carcinoma (CRC) cell lines. Methods: Human CRC cell lines Lovo and SW 480 were cultured. Three pairs of small interfering RNA (siRNA) targeting Ezrin were transfected into the CRC cells. Western blotting was used to detect the transfection rates so as to screen the most effective siRNA which had been transfected into the CRC cells. Real-time PCR and Western blotting were used to detect the downregulation of Ezrin mRNA and protein at different time points. We observed the biological behaviors of CRC cells when the expression level of Ezrin was the lowest. CCK-8 method was used to detect the proliferation of the cells. Flow cytometry was used to detect the cell cycle and apoptosis. Transwell test was used to detect the invasion and migration ability of the transfected CRC cells. Results: Real-time PCR and Western blotting results revealed that Ezrin siRNA notably down-regulated Ezrin expression at both mRNA and protein levels. Down-regulation of Ezrin expression distinctly decreased the proliferation rates of the two cell lines. After RNAi treatment, the cell proportion in S phase decreased slightly and the cell proportion in G1 phase increased slightly in the two cell lines (P > 0.05). The apoptotic index increased in the three siRNA transfected groups compared with the control group (P < 0.05). The migration and invasion ability of CRC cells significantly decreased after siRNA interference (P < 0.01). Conclusion: Ezrin plays an important role in the process of proliferation, migration, and invasion of CRC cells. It has the potential to become a new target to inhibit tumorigenesis and development of CRC.