MiR-93-5p Affects Cervical Cancer Cells Biological Behaviour Through Target TNFAIP1

Worldwide,the incidence of cervical cancer ranks second in female cancer.In China,new cases of cervical cancer would increase more than 11 million a year,and about 20,000 to 30,000 women die of cervical cancer a year.Although the treatment is applied to cervical cancer surgery combined with radiotherapy,chemotherapy and other advanced technology,but there isn’t one way to completely treat cervical cancer.It is mainly because we have no idea about the mechanism of cervical cancer progression.Currently,many miRNAs have been reported aberrantly expressed in cervical cancer and directly involve in cancer carcinogenesis and progression.For example,the expression of miR-497 is significantly lower in cervical cancer cells and cancer tissues than which in cervical epithelial cells and the adjacent normal tissue.Abnormal expression of miR-497 often give rise to a high risk of cancer metastasis.MiR-196a in squamous cervical cancer is abnormal high expression,which can negatively regulate netrin 4,and the expression of netrin 4 protein is associated with a invasive tumor and poor prognosis.These evidences suggested that miRNAs promote or inhibit cervical cancer development via various pathways and further affect biology behaviors.In this study,the expression of miR-93-5p in cervical cancer tissues and adjacent tissues was tested and analysed by QRT-PCR.The result showed that the expression of miR-93-5p in cervical cancer was significantly higher than the corresponding adjacent tissues.At the same time,we choose four types of cervical cancer cells(Hela,Siha,CaSki,C-33A)and an endometrial cancer cell(HEC-1 A),in which miR-93-5p were tested by QRT-PCR analysis.We found that miR-93-5p is highest expression in CaSki,lower expression in Hec-la.So we choose this tow types of cell as object of study.We transfected with miR-93-5p to HEC-1A and miR-93-ASO to HEC-1A to study miR-93-5p effects on HEC-1 A and CaSki proliferation,migration and invasion.The results showed that overexpression of miR-93-5p can enhance the proliferation,migration and invasion of of HEC-1 A cells.In contrast,miR-93-ASO inhibits the proliferation,migration and invasion of CaSki cells.The bioinformatics analysis results showed that TNFAIP1 may be a downstream target genes of mir-93-5p.TNFAIP1 is induced by tumor necrosis factor α(TNF-α)or interleukin 6(IL6),which plays an important role in TNFa signaling pathway and DNA synthesis or apoptosis.We detected the endogenous TNFAIP1 of five cell lines by Western bloting and the result shows that HEC-1A has the highest expression of endogenous TNFAIP1 and CaSki has lowest one.Luciferase assay proved that TNFAIP1 can be regulated by miR-93-5p.The results of Western blotting and QRT-PCR have proved that mir-93-5p can inhibit the expression of TNFAIP1in transcription and protein levels.On the other hand,we detected TNFAIP1 expression in cervical cancer tissues and normal tissues by QRT-PCR.The results showed that TNFAIP1 has a lower expression in cervical cancer tissues than which in adjacent tissues.Compared with control,HEC-1 A transfected with si-TNFAIP1 had a higher proliferative ability and stronger capability for metastasis.Instead,over expression TNFAIP1 dramaticly inhibited the CaSki ability of proliferation and metastasis.These result consistents with the effect of miR-93-5p or miR-93-ASO on cervical cancer cells.Thus,our research results demonstrated miR-93-5p regulates cell growth and metastasis of cervical cancer cells.Meanwhile,it is possible provided a new proof for leading to cervical cancer carcinogenesis through miRNAs signal pathway.Most important of all,it provides theoretical basis for disclosure muti-regulation mechanism of cervical cancer and for treatment of cervical cancer and its related diseases through genetic diagnosis and ultimately use of molecular interventions in the future.