抑癌基因WTX对胃癌SGC-7901细胞增殖、凋亡及细胞周期的影响

目的:探讨WTX基因对人胃癌SGC-7901细胞生物学行为的影响。方法:将WTX重组质粒或空载体质粒用Attractene法转染SGC-7901细胞,以无处理SGC-7901细胞为空白对照,检测不同时间e GFP标记的转染效率;RT-PCR法检测WTX m RNA水平;CCK-8法测定细胞增殖情况;流式细胞技术检测转染效率、凋亡及细胞周期的变化。结果:转染WTX基因48 h后,e GFP表达最强,转染效率达(33.10±4.16)%;与空白对照组和空载体组比较,WTX转染组WTX m RNA表达明显升高;细胞增殖能力明显降低,S期细胞明显增多,而G1期和G2/M期细胞减少(均P<0.05)。各组细胞均未见明显的细胞凋亡。结论:WTX可通过诱导S期阻滞抑制胃癌细胞SGC-7901生长,但不影响细胞凋亡。

Influence of tumor suppressor WTX gene on proliferation, apoptosis and cell cycle of human gastric cancer SGC-7901 cells

Objective: To investigate the influence of WTX gene on biological behaviors of human gastric cancer SGC-7901 cells. Methods: The recombinant plasmids bearing WTX gene or empty plasmid vectors were transfected into SGC-7901 cells by using Attractene reagent, and the untreated SGC-7901 cells were used as blank control. The EGFP-tagged transfection efficiency at different transfection times was determined, the WTX mRNA expression was measured by RT-PCR method, the cell proliferation was detected by CCK-8 assay, and the apoptosis and cell cycle were analyzed by flow cytometry. Results: The strongest expression of eGFP presented at 48 h after WTX gene transfection, when the transfection efficiency reached (33.10±4.16) %. In SGC-7901 cells of WTX transfection group compared with either blank control group or empty vector group, the WTX mRNA expression was increased significantly, proliferative ability was decreased significantly, and the number of S-phase cells was increased while the number of G1- and G2/M-phase cells was decreased significantly (all P<0.05). There was no significant apoptosis in any of the groups of cells. Conclusion: WTX gene can inhibit proliferation through inducing S-phase arrest in SGC-7901 cells, but has no influence on cell apoptosis.