Impact of siRNA-mediated down-regulation of CD147 on human breast cancer cells
- VernacularTitle:沉默 CD147基因对人乳腺癌 MDA－MB－231细胞的影响
- Author:
Zhenqian LI
1
;
Daoming LI
;
Jiangwei LI
;
Pei HUANG
;
Hui QIN
Author Information
1. 450052,郑州大学第一附属医院病理科 河南省肿瘤病理重点实验室
- Keywords:
Breast neoplasms;
Cell line,tumor;
Antigens,CD147;
Matrix metalloproteinase 2;
Tissue inhibitor of metalloproteinases
- From:
Chinese Journal of Pathology
2015;(10):734-738
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the influence of siRNA-mediated down-regulation of CD147 on growth, proliferation and movement of human breast cancer cell line MDA-MB-231.Methods The protein expression of CD147, MMP-2 and TIMP-2 of the MDA-MB-231 cells were analyzed by ABC.Lentiviral expression vector of CD 147 gene was constructed and transfected into MDA-MB-231 cells.RT-PCR and Western blot were used to detect the mRNA and protein level changes of CD 147 genes to identify the optimal time point , followed by detection of changes of mRNA and protein expression of MMP-2 and TIMP-2 genes. CCK-8 reagent method and cell scratch test were used to detect the proliferation and migration change of MDA-MB-231 cells.The nude mouse model of breast cancer by hypodermic injection with MDA-MB-231 cells was established to document the effect of CD 147 siRNA on the tumor transplants.Results After transfection of lentiviral expression vector of CD 147 gene, protein of CD147, MMP-2 and TIMP-2 were weakly or negative expressed , significantly weaker than those of control group ( P<0.01 ) .After 72 hours of transfection, average down-regulation rate of CD147 and MMP-2 were 96.03%±0.84% and 96.03%± 0.84%, respectively.Both CD147 mRNA and MMP-2 mRNA expression were down-regulated(P<0.05), while TIMP-2 mRNA expression showed no significant deference ( P >0.05 ) .No less than 2 days after transfection, cell growth of MDA-MB-231 cell line was found significantly inhibited ( P <0.05 ) .After 24 hours of tranfection, average migration distance of MDA-MB-231 cell line and control group were (0.64 ± 0.12)mm and (4.69 ±0.85)mm, respectively, which indicated a lower migrate speed.Down regulation of CD147 led to reduction of volume and mass of nude mouses .The growth of the carcinoma transplant was inhibited upon siRNA-mediated down-regulation of CD147 ( P <0.05 ) , with an average tumor mass of (1.85 ±0.98) g and both reduction of tumor size and tumor mass .Conclusions CD147 may alter the MMP-2/TIMP-2 balance in MDA-MB-231 cells. CD147 gene silencing inhibits the proliferation and migration of MDA-MB-231 cells and the growth of carcinoma transplants in nude mice .
