Expression And Correlation Of MiR-135a-5p,GATA3 And STAT3 In Cervical Cancer

Objectives To investigate the expression of miR-135a-5p,GATA3 and STAT3 in different pathological types of cervical tissues and cervical cancer cell lines,and analyze the correlation between the expressions of miR-135a-5p and GATA3 mRNA,and the correlation between GATA3 and STAT3 mRNA,to investigate the effect of miR-135a-5p targeting GATA3 on STAT3 expression in cervical cancer cells,and analyze the correlation between miR-135a-5p and GATA3 mRNA expression in cervical cancer patients with clinicopathological parameters.Methods A total of 155 cervical specimens were collected from the Tangshan Gongren Hospital from September 2014 to September 2017.The specimens were selected according to the inclusion and exclusion criteria of the specimens.The specimens were confirmed by the experimental staff and the hospital pathologists.The grades and types included 37 patients in the normal control group and 81 patients in the cervical intraepithelial neoplasia(CIN)group(39 in the CINI group,42 in the CINII-III group)and 37 in the cervical cancer(CSCC)group.RT-qPCR was used to detect miR-135a-5p,GATA3 and STAT3 mRNA in normal cervical(normal control)and cervical cancer(CSCC)tissues,human normal cervical epithelial cells(HcerEpic),HeLa and SiHa cervical cancer cells.The expression levels of miR-135a-5p,GATA3 and STAT3 mRNA in cervical cancer tissues was correlated.The expression of GATA3 and STAT3 in different pathological types of cervical tissues was detected by immunohistochemistry.HcerEpic was detected by Western Blot.Protein expression of GATA3 and STAT3 in HeLa and SiHa cells;Dualluciferase reporter gene verification of miR-135a-5p and GATA3 3' untranslated region(UTR);knockdown of miR-135a-5p with inhibitor in HeLa cells,then use RT-qPCR assay detect the expression of GATA3 and STAT3 mRNA,Western Blot assay detect the expression of GATA3 and STAT3 protein level;the relationship between the expression of miR-135a-5p and GATA3 mRNA with clinical pathological parameters of cervical cancer was analyzed.Results 1 RT-qPCR showed miR-135a-5p was significantly up-regulated in cervical cancer tissues and cells(P<0.01,P<0.05,P<0.05);GATA3 mRNA expression level: compared with normal control group,the expression of GATA3 mRNA in CINII-III and CSCC group were statistically significant(P<0.05);The expression of STAT3 mRNA increased with the degree of cervical lesions(P<0.05).Immunohistochemistry experiment showed with the progression of cervical lesions,GATA3 positive cells showed a decreasing trend,while STAT3 positive cells increased significantly.Western Blot showed that compared with HcerEpic,GATA3 protein was lowly expressed in cervical cancer cells,while STAT3 protein was over-expressed in cervical cancer cells,the differences were statistically significant(P<0.05).2 Pearson correlation analysis showed that there was a negative correlation between miR-135a-5p and GATA3 mRNA expression in cervical cancer tissues(r=-0.6656,P<0.05);GATA3 was negatively correlated with STAT3 mRNA expression(r =-0.4534,P < 0.05).3 Using TargetScan to predict the target gene of miR-135a-5p,which showed that miR-135a-5p has a binding site with GATA3 3'UTR,and Dual-luciferase-reporter gene further verified that miR-135a-5p can targeted the 3'UTR of GATA3 to inhibit the transcriptional activity of GATA3.4 Knockdown the expression of miR-135a-5p with inhibitor,4 Knockdown the expression of miR-135a-5p with inhibitor,RT-qPCR and Western Blot assay showed that GATA mRNA and protein expression levels was up-regulated in miR-135a-5p inhibitor group,while STAT3 mRNA and protein expression levels was down-regulated,the difference was statistically significant.,the difference was statistically significant.5 Correlation analysis of clinical and pathological parameters of cervical cancer confirmed that the expression of miR-135a-5p in cervical cancer tissues was correlated with clinical stage and lymph node metastasis(P<0.05);GATA3 mRNA expression was correlated with clinical stage,depth of interstitial invasion and lymph node metastasis(P<0.05).Conclusions 1 In cervical cancer,the expression of miR-135a-5p was up-regulated,the expression of GATA3 was down-regulated,the expression of STAT3 was up-regulated.2 miR-135a-5p can inhibit the transcriptional activity of GATA3,which in turn activates STAT3,then promote the occurrence and development of cervical cancer.3 Knocking down the expression of miR-135a-5p in cervical cancer cells would abolish the inhibition of GATA3 transcriptional activity and reduce the expression of STAT3,which may become a new target for targeted therapy.Figure 9;Table 5;Reference 116.