Effect Of MiR-205on Biological Behavior In Ovarian Cancer Cells

Background and objective:Ovarian cancer is one of the three female reproductive system cancers and second leading cause of death worldwide among gynecological cancers.Because ovarian cancer is asymptomatic in the early stages and there are no efficient tumor specific and sensitive markers to monitor epithelial ovarian cancer,in most women, ovarian cancer is difficult to treat with a five year survival rate of around20%in cancers diagnosed in advanced stage.In recent years,miRNAs regulating gene expression has become hot spot in cancer research.miRNAs are small(19-25ribonucleotides),single-stranded non-coding RNAs found in plant,invertebrate and vertebrate genomes.miRNAs are involved in regulating cell differentiation,cell cycle progression and apoptosis.More than900miRNAs have been identified in human to date.miR-205,located in1q32.2,is a piece of uccuucauucc-accggagucug encoded microRNA, showing highly conserved in different kinds of lines.miR-205may function as an oncogene or tumor suppressor gene depending on the cellular contexts.Consistent with its dual role,several studies have demonstrated its tumor promoting and suppressive roles in different cancer cell lines.Our previous comparative proteomics studies have found that two differential expression proteins Lamin A/C and Ezrin are highly related to the invasion and metastasis in ovarian cancer.Meanwhile,we have predicted the existence of potential binding sites of miR-205in the3’UTR of Lamin A/C and Ezrin by bio-informatics (three independent software TargetScan, DIANA-miroT, PicTar).So we speculate that there may be a close relationship between miR-205and the two differentially expressed proteins in ovarian cancer.This paper aims to explore the effect of miR-205in ovarian cancer cell proliferation,migration and invasion,and the relationship between miR-205and the two differentially expressed proteins Lamin A/C and Ezrin.Methods:(1)Four human ovarian cancer cell lines were cultured under sameconditions:HO-8910(low-metastatic),HO-8910pm(high-metastatic), SKOV-3(low-metastatic),SKOV-3ip(high-metastatic).(2)The miR-205expression in four cell lines was detected by quantitative real time PCR.(3)The miR-205mimics/inhibitor was transiently transfected into the four cell lines.The miR-205expression in the transfected cell lines was detected by quantitative real time PCR.(4)The migration and invasion effects on transfected cell lines were detected by Transwell migration assay and invasion assay.(5)Proliferation of the transfected cell lines were detected by MTT assay.(6)The Cell Clonies of the transfected cell lines were detected by Clonogenic assay.(7)Lamin A/C and Ezrin expression with the transfected cell lines were detected by Western blot.Results:(1)Under the same culture condition,the number of seeding and culture time,the cells of HO-8910pm and the SKOV-3ip showed faster growth than that of HO-8910and SKOV-3.(2) The expression of miR-205in the transfected HO-8910pm and SKOV-3ip were higher than that of HO-8910and SKOV-3.(3)The expressions of miR-205in the transfected HO-8910and SKOV-3were higher than miR-205control(P<0.05);the expressions of miR-205in the transfected HO-8910pm and SKOV-3ip were lower than miR-205control(P<0.05);(4) The migration and invasion ability in the transfected HO-8910and SKOV-3were higher than miR-205control(P<0.05);the migration and invasion ability in the transfected HO-8910pm and SKOV-3ip were lower than miR-205control(P<0.05);(5) The proliferation ability in the transfected HO-8910was higher than miR-205control(P<0.05);The proliferation ability in the transfected SKOV-3was higher than miR-205control,but there was no statistically significant difference with the miR-205control(P>0.05);the proliferation ability in the transfected HO-8910pm and SKOV-3ip were lower than miR-205control,but there were no statistically significant differences with the miR-205controls(P>0.05).(6)The number of clonies in the transfected HO-8910and SKOV-3were more than miR-205control(P<0.05);the number of clonies in the transfected HO-8910pm and SKOV-3ip were less than miR-205control(P<0.05);(7)The protein level of Lamin A/C and Ezrin were lower in the transfected HO-8910and SKOV-3than the miR-205control;The protein level of Lamin A/C and Ezrin were higher in the transfected HO-8910pm than the miR-205control;there were no significant effects on SKOV-3ip.Conclusion:(1)miR-205can enhance the proliferation,migration and invasion ability in ovarian cancer.(2)miR-205can down-regulate the levels of Lamin A/C and Ezrin protein.(3) The promotion of OVCA cells invasion by miR-205may partially due to the regulation of Lamin A/C and Ezrin.