Expression Of MiR-196a Target Gene HoxB8 In Cervical Cancer Cells And Its Clinical Significance

Objective This study is to investigate the expression of miR-196 a and HoxB8 mRNA in different cervical cancer cell lines,affection of HoxB8 gene to the apoptosis of cervical cancer cells,and the expression and clinical significance of HoxB8 in cervical cancer.Method 1.The expression of miR-196 a and HoxB8 mRNA in cervical cancer Caski,SiHa,C33-A and HeLa cell lines were detected by real-time quantitative PCR;The effection of the miR-196 a simulation on HoxB8 expression was detected by qRT-PCR;the activity of Caspase3/7 was detected after the HoxB8 siRNA was transfected to cervical cancer SiHa cells;2.Clinical and pathological data of 67 patients with HPV16 positive IIb-IIIb stage cervical squamous carcinoma in department of Gynecologic Oncology of Affiliated Tumor Hospital of Guangxi Medical University between February 2011 to December 2012 was collected.Through immunohistochemical staining method,We check expression of HoxB8,and then further analyze the correlation between immunohistochemical results,clinicopathologic factors and prognosis.Survival rate was calculated with Kaplan-Meier method.TheLog-rank method was used for comparing survival curves.Correlation between clinicopathologic factors was analysed with chi-square or Fisher's exact test.Cox regression model was used for multiple factors analysis.Results 1.Real-time quantitative PCR showed that miR-196 a expression in HPV16 positive Caski and SiHa cell lines was significantly lower than that in HPV16-negative C33-A and HeLa,the difference was statistically significant(P<0.001).Correspondingly,the expression of HoxB8 in HPV16-positive Caski and SiHa cell lines was significantly higher than that in HPV16-negative C33-A and HeLa,the difference was statistically significant(P<0.001).Overexpression of miR-196 a significantly reduced HoxB8 expression.Capase3/7 activity array results showed that Capase3/7 activity in Si Ha cell HoxB8 siRNA transfer group was significantly higher than the negative control group and normal control group,and the difference was statistically significant(P < 0.05).2.For HoxB8,10 samples presented negative expression while 57 samples presented positive espression(85%).The correlation analysis showed that HoxB8 had no significant statistical significance with age,tumor size,preoperative SCC,differentiation,and FIGO stage.In 67 cases,the median overall survival time was 48 months,1 year,3 years and 5 years overall survival rate were 91%,66% and 54% respectively.Single factor and multi-factor survival analysis showed that tumor diameter>4cm,pure radiotherapy were the independent risk factors for the prognosis of patients with IIb-IIIb stage cervical cancer,the difference was statistically significant(P < 0.05).Conclusion MiR-196 a may through the downstream target gene HoxB8 affect cervical cancer cell apoptosis.There was no significant correlation between HoxB8 and IIb-IIIb stage cervical squamous carcinoma patient age,tumor size,preoperative SCC,differentiation,and FIGO stages.The tumordiameter>4cm and Pure radiation therapy in patients with OS shortened,tumor diameter and therapy method were independent prognostic factors of OS.There was no obvious correlation between HoxB8 and prognosis of IIb-IIIb stage cervical squamous carcinoma patients.