RNA干扰IDH2基因对HCT-8细胞增殖能力的影响

J4 ›› 2012, Vol. 38 ›› Issue (4): 649-652.

RNA干扰IDH2基因对HCT-8细胞增殖能力的影响

徐晓芳¹|宋晗星^*|邢沈阳¹|吕强²|高江明²|赵娜²|常乐²|高峰¹|宫鹏涛²|李建华²|张国才²|张西臣²

(1.吉林大学畜牧兽医学院动物科学系|吉林长春130062；2.吉林大学畜牧兽医学院预防兽医系|吉林长春130062)

收稿日期:2012-04-11 出版日期:2012-07-28 发布日期:2012-07-28
通讯作者: 邢沈阳 E-mail:(Tel：0431-87835138，E-mail：xingsy@yahoo.com.cn)
作者简介:徐晓芳(1986-)|女|辽宁省辽阳市人|在读动物科学硕士|主要从事肿瘤遗传学研究。
基金资助:
国家“973”项目资助课题(2006CB910505);吉林省科技厅科技发展计划项目资助课题(20101585，2003055025，20106044)

Influence of IDH2 on proliferation of coloncancer cells HCT-8 by siRNA

XU Xiao-fang¹| SONG Han-xing^*, XING Shen-yang¹, LV Qiang², GAO Jiang-ming², ZHAO Na², CHANG Le², GAO Feng¹, GONG Peng-tao², LI Jian-hua², ZHANG Guo-cai², ZHANG Xi-chen²

(1.Deprtment of Animal Science|College of Animal Science and Veterinary Medicine, Jilin University,Changchun 130062, China；2.Deparment of |Preventive Veterinary Medicine|College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China)

Received:2012-04-11 Online:2012-07-28 Published:2012-07-28

摘要/Abstract

摘要：

目的：研究RNA干扰(RNAi)对结肠癌HCT-8细胞IDH2基因表达的抑制作用及干扰后对HCT-8细胞增殖的影响，为结肠癌的基因靶向治疗提供理论依据。方法：实验分siRNA干扰质粒转染组、阴性质粒对照组和HCT-8细胞空白对照组。构建靶向基因IDH2的siRNA真核绿色荧光载体重组质粒PGPU6/GFP/Neo-IDH2，利用FuGENE HD转染剂转染HCT-8细胞，通过荧光定量PCR检测IDH2 mRNA的表达情况及MTT法检测结肠癌HCT-8细胞增殖变化。结果：在荧光显微镜下进行细胞计数，细胞转染率为74%。实时荧光定量PCR检测，siRNA重组质粒PGPU6/GFP/Neo-IDH2对IDH2基因表达的抑制率为85.02%，与空白对照组比较差异有统计学意义(P<0.05)。MTT检测，干扰质粒转染组细胞的增殖能力明显低于空白对照组(P<0.05)。结论：抑制IDH2基因的表达可影响结肠癌细胞的增殖能力，提示IDH2基因在结肠癌的发生发展中起重要作用，抑制IDH2基因的表达可能成为一种治疗结肠癌的方法。

关键词: RNA干扰；IDH2基因；人结肠癌HCT-8细胞

Abstract:

Abstract:Objective
To analyze the inhibitory effect of RNA interference (RNAi) on the expression of IDH2 gene of colon cancer cells HC７-8 and the influnce in the proliferation of colon cancer cells in vitro,and to provide a theoretical basis for colon cancer targeted therapy.Methods The experiment was divided into siRNA plasmid transfection,negative plasmid control and HCT-8 cells control groups.The recombinant plasmid which contained the siRNA of the IDH2 gene in the green fluorescence eukaryotic vector (PGPU6/GFP/Neo-IDH2) and the negative control were constructed and then they were transfected into colon cancer cells by using FuGENE HD Transfection Reagent. The expression of IDH2 gene mRNA was detected by real-time PCR. The proliferation abilities of colon cancer cells were detected by MTT.Results Counted by fluorescence microscope,the transfection rate of cells was 74%. The inhibitory rate of IDH2 mRNA in PGPU6/GFP/Neo-IDH2 group was 85.02%,which had significant difference compared with HCT-8 cells control group (P<0.05). Compared with neagtive control group,the proliferation ability of colon cancer cells was deceased by MTT assay (P<0.05).Conclusion Inhibition of the expression of IDH2 gene can infect the proliferation ability of colon cancer cells. It shows that IDH2 gene plays a role in the development of colon cancer and it may be beneficial in finding a new therapy for colon cancer.

Key words: RNA interference, IDH2 gene, colon cancer cells HCT-8

中图分类号:

徐晓芳|宋晗星|邢沈阳|吕强|高江明|赵娜|常乐|高峰|宫鹏涛|李建华|张国才|张西臣. RNA干扰IDH2基因对HCT-8细胞增殖能力的影响[J]. J4, 2012, 38(4): 649-652.

XU Xiao-fang, SONG Han-xing, XING Shen-yang, LV Qiang, GAO Jiang-ming, ZHAO Na,C. Influence of IDH2 on proliferation of coloncancer cells HCT-8 by siRNA[J]. J4, 2012, 38(4): 649-652.

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