沉默MYCT-1基因表达对人胃粘膜细胞系GES-1增殖凋亡的影响

Effect of Silencing MYCT-1 Gene Rxpression on Proliferation and Apoptosis of Human Gastric Mucosal Cell Line GES-1

为了探讨沉默MYCT-1基因表达对人胃粘膜细胞系GES-1细胞增殖和凋亡的影响，本研究将人胃粘膜GES-1细胞系分为空白组（GES-1细胞系培养）、沉默组（加入2μL 2μmol／LMYCT-1特异性RNA与GES-1细胞系共培养）和对照组（加入2μL 2μmol／LsiRNANC与GES-1细胞系共培养）。采用RT-PCR技术于培养48h后测量各组MYCT-1mRNA的表达水平；采用Westernblotting技术检测各组MYCT-1蛋白的表达水平；采用流式细胞仪技术检测各组细胞分期构成、MTT法测定细胞增殖、AnnexinV-FITC双染法检测细胞凋亡情况。研究显示：沉默组G0／G1期和G2／M期细胞所占比例显著低于对照组和空白组（p〈0．05），沉默组S期细胞所占比例显著高于对照组和空白组（p〈0.05）；对照组和空白组的G0／G1期、G2／M期和S期细胞所占比例差异均不显著（p〉0.05）；沉默组细胞凋亡率显著高于对照组和空白组（p〈0．05），沉默组细胞增殖率显著低于对照组和空白组（p〈0.05）；对照组和空白组的细胞凋亡率和增值率差异均不显著（p〉0．05）；沉默组MYCT-1 mRNA、MYCT-1蛋白显著低于对照组和空白组（p〈0．05）；对照组和空白组的MYCT-1 mRNA和MYCT-1蛋白差异均无统计学意义（p〉0．05）。沉默MYCT-1基因表达可显著促进GES-1细胞凋亡，抑制GES-1细胞增殖作用。

In order to investigate the effect of silencing MYCT-1 gene expression on the proliferation and apoptosis of human gastric mucosal cell line GES-1. The human gastric mucosa GES-1 cells were divided into blank group （GES-1 cell culture）, silent group （adding 2 μL 2 μmol/L MYCT-1 specific RNA and co-cultured with GES-1 cells） and control group （adding 2 μL 2 μmol/L siRNA into NC and co-cultured with GES-1 cell lines）. The mRNA expression level of MYCT-1 were measured by RT-PCR technology after cultured for 48 h. The expression level of MYCT-1 protein was detected by Western blotting. The cell structure of each group was detected by flow cytometry. Cell proliferation and Annexin V-FITC double staining were used to detect apoptosis. The results showed that the proportion of G0/G1 phase and G2/M phase cells in silence group was significantly lower than that in control group and blank group （p 〈0.05）. The proportion of S phase cells in silent group was significantly higher than that in control group （/9〈0.05）. There was no significant difference in the proportion of G0/G1 phase, G2/M phase and S phase cells between the control group and the blank group （p 〉0.05）; The cell proliferation rate of the silent group was significantly lower than that of the control group and the blank group （p〈0.05）. There was no significant difference in apoptosis rate and proliferation rate between the control group and the blank group （p〉0.05）. The levels ofMYCT- I mRNA and MYCT- 1 in the silencing group were significantly lower than those in the control group and the blank group （p〈0.05）. There was no significant difference in MYCT-1 mRNA and MYCT-1 protein between the control group and the blank group （p 〉0.05）. Silencing MYCT-1 gene expression can significantly enhance the apoptosis ofGES- 1 cells and inhibit the proliferation of GES- 1 cells.