错配修复基因MSH2再表达对雌激素诱导结肠癌LOVO细胞凋亡的影响  

Effects of MSH2 gene re-expression on estrogen induced-apoptosis of colon cancer cells LOVO

作　　者：吕晨曦[1] 王德智[2] 金鹏[2] 何玉琦[2] 李爱琴[2] 杨欣艳[2] 盛剑秋[2] 

机构地区：[1]大连医科大学 [2]北京军区总医院消化内科,100700

出　　处：《中华消化杂志》2014年第6期388-391,共4页Chinese Journal of Digestion

基　　金：国家自然科学基金（81201958）

摘　　要：目的 观察错配修复基因MSH2再表达对雌激素诱导结肠癌LOVO细胞凋亡的影响并探讨其机制.方法 按转染质粒不同及是否进行雌二醇干预将结肠癌LOVO细胞分为空质粒十乙醇组、空质粒十雌二醇组、MSH2＋乙醇组、MSH2＋雌二醇组、雌激素受体（ER）β＋乙醇组、ERβ＋雌二醇组、ERβ＋ MSH2＋乙醇组、ERβ＋ MSH2＋雌二醇组并予相应处理.用Western印迹法检测ERβ、MSH2的转染效率和凋亡相关caspase3蛋白的表达情况.用细胞计数试剂盒8法检测各组细胞的活性.用凋亡DNA碎片提取试剂盒提取各组细胞的DNA碎片并观察DNA碎片的梯状条带.用流式细胞仪检测各组细胞的凋亡率.多组间比较行单因素方差分析,两组间比较行t检验.结果 转染后,LOVO细胞中ERβ和MSH2蛋白的表达水平均显著升高,且其表达均不受雌二醇的影响.ERβ＋雌二醇组和ERβ＋ MSH2＋雌二醇组caspase3蛋白的裂解活化片段表达水平高于其他组,但该二组之间则无明显差异.空质粒十乙醇组、空质粒十雌二醇组、MSH2＋乙醇组、MSH2＋雌二醇组、ERβ＋乙醇组、ERβ＋雌二醇组、ERβ＋ MSH2＋乙醇组、ERβ＋ MSH2＋雌二醇组的LOVO细胞活性分别为1.72±0.25、1.74±0.31、1.77±0.35、1.74±0.33、1.70±0.34、1.02±0.48、1.71±0.31、1.07±0.18,组间差异有统计学意义（F=3.791,P＜0.05）;其中ERβ＋雌二醇组低于ERβ十乙醇组,ERβ＋ MSH2＋雌二醇组低于ERβ十MSH2＋乙醇组,ERβ＋雌二醇组低于空质粒十雌二醇组,ERβ＋ MSH2＋雌二醇组低于MSH2＋雌二醇组,差异均有统计学意义（t=3.158、3.075、3.648、3.253,P均＜0.05）.ERβ＋雌二醇组和ERβ＋MSH2＋雌二醇组中可见细胞凋亡后DNA碎片形成的DNA梯状条带.空质粒十乙醇组、空质粒十雌二醇组、MSH2＋乙醇组、MSH2＋雌二醇组、ERβ＋乙醇组、ERβ＋雌二醇组、ERβ＋MSH2＋乙醇组、ERβ＋MSH2＋雌二醇组的细胞凋亡率分别为7.86±0.19、7.8Objective To observe the effects of MSH2 gene re expression on estrogen-induced apoptosis of colon cancer cells LOVO,and to explore its mechanisms.Methods According to different plasmid and whether with estradiol intervention,colon cancer LOVO cells were divided into empty plasmid with ethanol group,empty plasmid with estradiol group,MSH2 with ethanol group,MSH2 with estradiol group,estrogen receptor （ER） β with ethanol group,ERβ with estradiol group,ERβ with MSH2 and ethanol group and ERβ with MSH2 and estradiol group,and received corresponding treatment.The expression of MSH2,ERβ protein and apoptosis related caspase 3 protein were detected by Western blotting.Cell viability was measured by cell counting kit-8.Cell DNA fragments of each group were isolated with apoptosis DNA fragments isolation kit.And the DNA ladder was observed.The rate of apoptosis was detected by flow cytometer.Single factor variance analysis was performed for comparison among multiple groups,and t test was used for comparison between the two groups.Results After transfection,the expression of the MSH2 and ERβ at protein level in LOVO cells significantly increased and neither of their expression was effected by estradiol.The expression levels of caspase 3 cleavaged active fragments of ERβ with estradiol group and ERβ with MSH2 and ethanol group were higher than other groups,and there was no significant difference between these two groups.The LOVO cell viability of empty plasmid with ethanol group,empty plasmid with estradiol group,MSH2 with ethanol group,MSH2 with estradiol group,ERβ with ethanol group,ERβ with estradiol group,ERβ with MSH2 and ethanol group and ERβ with MSH2 and estradiol group was 1.72 ±0.25,1.74 ± 0.31,1.77 ± 0.35,1.74±0.33,1.70±0.34,1.02±0.48,1.71±0.31 and 1.07±0.18,respectively,and the differences between the groups were statistically significant （F=3.791,P＜0.05）.Among them,the LOVO cell viability of ERβ with estradiol group was lower than that of ERβ with ethanol group,accordingly,that

关 键 词：结肠肿瘤 雌激素类 雌激素受体β 细胞凋亡 MSH2蛋白 

分 类 号：R735.35[医药卫生—肿瘤]