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

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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. (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

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

CLC Number: 

  • Q78