Hsa-miR-218 inhibits growth of cervical cancer HeLa cells by targeting to LASP1
10.3969/j.issn.1000-4718.2015.09.007
- VernacularTitle:Hsa－miR－218靶向调控 LASP1对宫颈癌 HeLa 细胞生长的影响
- Author:
Yu QIU
;
Jianping HUANG
;
Qinxian ZHOU
;
Huan WANG
;
Huigang SHI
;
Jinhua HE
- Publication Type:Journal Article
- Keywords:
Hsa-miR-218;
LIM and SH3 protein 1;
Cervical cancer;
HeLa cells
- From:
Chinese Journal of Pathophysiology
2015;(9):1572-1577
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To study the effect of hsa-miR-218 on cervical cancer HeLa cell growth and the underlying molecular mechanism .METHODS:The lentivirus expression vector pmiR-218 targeting to hsa-miR-218 was constructed . pmiR-218 was transfected into HeLa cells .The number of viable HeLa cells was counted by the method of Trypan blue ex-clusion.The inhibitory rate of cell activity was detected by WST-8 assay.The expression of LIM and SH3 protein 1 (LASP1) at mRNA and protein levels was determined by real-time PCR and Western blot.The interaction between miR-218 and LASP1 was examined using a luciferase reporter assay .RESULTS:The lentivirus expression vector pmiR-218 tar-geting to hsa-miR-218 was constructed successfully and confirmed by DNA sequencing .Over-expression of miR-218 inhibi-ted the activity of HeLa cells with the inhibitory rates of 15%, 26%and 65%at 24 h, 48 h and 72 h, respectively .The difference between transfection group and blank control /negative control group was statistically significant .The luciferase activity was reduced when co-transfection with miR-218 mimics and LASP1-3’ UTR plasmid.The relative expression of miR-218 was increased after transfection with pmiR-218.Over-expression of miR-218 down-regulated the LASP1 expression at mRNA and protein levels by 25%and 75%respectively.Compared with blank control group and negative control group , the difference was statistically significant (P<0.05).CONCLUSION:pmiR-218 effectively inhibits the growth of HeLa cells in a time-dependent manner.miR-218 targets to the 3’UTR of LASP1, thus down-regulating the expression of LASP1 in HeLa cells .
