RNA干扰PLCE1基因表达对食管鳞癌细胞周期和凋亡的影响

摘要

目的：通过siRNA干扰PLCE1基因表达，检测PLCE1对食管鳞癌细胞增殖周期和凋亡的影响，以探讨PLCE1的致癌机制。方法：采用免疫组织化学方法检测食管鳞癌组织和癌旁正常组织中PLCE1蛋白的表达水平；PLCE1特异性siRNA转染食管鳞癌细胞，荧光显微镜观察转染效率；半定量RT-PCR法检测转染后PLCE1沉默效果；流式细胞术检测干扰后细胞周期变化及凋亡情况。结果：PLCE1在食管鳞癌组织的表达高于正常组织（P=0.000）；siRNA转染后PLCE1表达明显减少（P=0.000）；与对照组相比，PLCE1干扰后可致G0/G1期细胞阻滞（P=0.001），细胞凋亡增多（P=0.000）。结论：PLCE1在食管鳞癌组织中有较高表达；沉默PLCE1后可抑制癌细胞的增殖，促进其凋亡。%Objective:To investigate the effects of PLCE1(phospholipase Cε1)gene silencing by siRNA(small in-terfering RNA)interference on the cell cycle and apoptotic of esophageal squamous carcinoma cells( ESCC),to identi-fy the mechanism. Methods:Esophageal cancer tissue specimens and normal esophageal mucosa were detected by im-munohistochemical staining assay to confirm overexpression of PLCE1 in esophageal carcinoma tissues. ESCC 9706 were transfected with the specific siRNA of PLCE1. Fluorescence microscopy was used to examine transfection effi-ciency,while the result of PLCE1 silencing was examine by reverse transcription( RT)-PCR. Flow cytometry and An-nexin-V apoptosis assay were used to detect the cell cycle and apoptosis,respectively. Results:The rate of PLCE1 expression in tumor tissue was significantly higher than the normal tissue(P=0. 000). After transfection,mRNA ex-pression of PLCE1 in ESCC 9706 was significantly reduced( P=0. 000). Compared with the control group,flow cy-tometry results suggested that the down-regulation of PLCE1 can arrested cell cycle in G0/G1 phase(P=0. 001);apoptosis was significantly higer than that of the control group(P=0. 000). Conclusion:PLCE1 showed significantly higher expression in esophageal carcinoma tissue;down-regulating PLCE1 could arrest the cell cycle and promote the apoptosis of ESCC.