Effect of apoptosis and proliferation of endometrial cancer HEC-1B cell by siRNA down-regulating expression of Ezrin

Abstract

Objective To observe the changes of cell cycle, apoptosis and proliferation of endometrial cancer cells after the expression and down-regulation of Ezrin in endometrial cancer cells and to explore whether Ezrin may be a candidate gene for targeted therapy. Methods Endometrial cancer cells were from Shanghai Institute of Cell Research, of Chinese Academy of Medical Sciences in February 2017 and divided into blank control group and siEzrin group according to the intervention methods.Western blot and qRT-PCR was used to detect the expression of Ezrin protein and mRNA in endometrial cell lines.Small interfering RNA(siRNA) was used to transfect HEC-1B cell and down-regulate Ezrin.Cell cycle and apoptosis were detected by flow cytometry.MTT assay was used to detect multiplication. Results Western blot showed that Ezrin protein was expressed in Ishikawa(31.742±5.832)、HEC-1A(16.326±3.135)、HEC-1B(17.636±4.426) and KLE(14.862±5.109) and qRT-PCR showed that mRNA was expressed in Ishikawa (2.513±0.725), HEC-1A (1.655±0.692), HEC-1B (3.237±0.411) and KLE (0.962±0.235) cell lines, and expressed highest in HEC-1B cells (F=6.173, P<0.05; F=7.042, P<0.05). Flow cytometry assay showed that compared with blank control group less cells stayed in G1 phase and G2 phase, more stayed in S phase (t=3.118, P<0.05; t=5.435, P<0.05; t=3.332, P<0.05). The apoptotic rate of HEC-1B cells increased from (9.84±2.37)% to (17.64±5.96)%(t=8.963, P<0.01) after Ezrin was down-regulated.MTT assay showed that the proliferation of HEC-1B cells in 72 h and 96 h siEzrin transfection group was lower than that in blank control group (t=3.209, P<0.05; t=3.726, P<0.05). Conclusion Down-regulating of Ezrin may promote more endometrial cancer cells stay in S phase and promote apoptosis, inhibit proliferation, Ezrin may become target candidate gene in target therapy. Key words: Endometrial cancer; Ezrin protein; Cell cycle; Apoptosis; Proliferation