Effect of ADAM17-shRNA on proliferation of human breast cancer cell line MCF-7 in hypoxia
10.3969/j.issn.1671-8348.2017.01.004
- VernacularTitle:ADAM17-shRNA在缺氧条件下对人乳腺癌细胞MCF-7增殖的影响
- Author:
Guofu CHEN
;
Lijun WU
;
Xuepeng ZHANG
;
Zhun CAI
;
Xiangchao MENG
- Keywords:
RNA;
polymerase chain reaction;
hypoxia;
depolymerization metalloproteinases 17;
RNAi;
proliferation
- From:
Chongqing Medicine
2017;46(1):29-32
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of shRNA targeting a disintegrin and metalloproteinases 17(ADAM17)on the proliferation of human breast cancer MCF-7 cells in hypoxia environment.Methods The four specific ADAM17-shRNA se-quences aiming at ADAM17 were designed,transfected into MCF-7 cells by electroporation,and cultured in hypoxia environment. The experiment was divided into the control group (blank phosphate buffer solution,PBS),nonsense sequence group (transfected with ADAM17-shNC)and shRNA transfection group (transfected with ADAM17-shRNA,the highest silencing efficiency of shR-NA was selected for following experiments).Real-time PCR was used to detect the expression of ADAM17 mRNA.The prolifera-tion ability and cell cycle change of MCF-7 cells were detected by iCELLigence and flow cytometry (FCM),respectively.Results Compared with control group and nonsense sequence group,the four ADAM17-shRNA transfection groups all had the silence effect on ADAM17 gene expression (P <0.05 ),the difference was statistically significant(P <0.05 ),particularly shRNA1219 had the highest inhibitory rate (F =5.11,P <0.01 ).The cellular proliferation ability and cell growth speed in the shRNA transfection group were significantly decreased compared with the control group and nonsense sequence group (P <0.05).Most cells of shRNA transfection group remained in the G0/G1 phase (73.35 ± 2.45 ),which in the control group and nonsense sequence group was (62.56±2.35)and (62.68 ±1.20)respectively,the difference was statistically significant(P <0.05).The cell cycle progression was significantly delayed.Conclusion ADAM17-shRNA inhibits the proliferation of MCF-7 cells under hypoxic environment.
