基础研究(Basic Research)

抑癌基因WTX 对胃癌SGC-7901 细胞增殖、凋亡及细胞周期的影响

Published at: 2015年第24卷第4期

杨蔚 1,2 , 张毅 3 , 钟玲 4 , 李思齐 3 , 杨金伟 2 , 陈嘉勇 3
1 昆明医科大学,云南 昆明 650500
2 云南省第一人民医院 普外二科 ,云南 昆明 650032
3 昆明医科大学第二附属医院 急诊科,云南 昆明 650101
4 云南省第一人民医院 急诊ICU,云南 昆明 650032
通讯作者 嘉勇 陈 Email: littlemark800306@yahoo.com.cn
DOI: 10.3978/j.issn.1005-6947.10.3978/j.issn.1005-6947.2015.04.010
基金:
云南省卫生科技计划资助项目 2012NS046
云南省科技厅- 昆明医科大学联合专项基金资助项目 2010CD165
云南省应用基础研究重点资助项目 2011FA029
云南省应用基础研究面上资助项目 2013FZ295

摘要

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eGFP 标记的转染效率;RT-PCR 法检测WTX mRNA 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,eGFP 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 mRNA 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

Abstract

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 EGFPtagged 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.


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引用

引用本文: 蔚 杨, 毅 张, 玲 钟, 思齐 李, 金伟 杨, 嘉勇 陈. 抑癌基因WTX 对胃癌SGC-7901 细胞增殖、凋亡及细胞周期的影响[J]. 中国普通外科杂志, 2015, 24(4): 511-516.
Cite this article as: YANG Wei, ZHANG Yi, ZHONG Ling, LI Siqi, YANG Jinwei, CHEN Jiayong . Influence of tumor suppressor WTX gene on proliferation, apoptosis and cell cycle of human gastric cancer SGC-7901 cells [J]. Chin J Gen Surg, 2015, 24(4): 511-516.