MiR-32 Target PTEN Regulate EMT Progress Invasion And Metastasis Of Cervical Cancer Cells

Objective: To search the miRNAs that could bind to phosphatase and tensin homologue deleted on chromosome ten(PTEN) through bioinformatics analysis. Study the relationship between mi R-32 and PTEN.To investigate the molecular mechanism of mi R-32 targted PTEN on invasion and metastasis of cervical cancer Hela cells.Methods: In this study, through the analysis of bioinformatics software to search mi RNAs that can combine with PTEN gene and further analysis found that, among them, miR-32 and PTEN with the best specificity and stability. miR-32 is detected by qRT-PCR, and PTEN and Epithelial-Mesenchymal Transition(EMT) related proteins are detedted by Western blot in cervical carcinoma and carcinoma adjacent tissues. To analyze the relationship between miR-32 expression and clinic pathological characters of cervical cancer. Verification of miR-32 and PTEN binding site: synthesis of miR-32 mimics, inhibitors, negative control(NC), and then transfected into Hela cells, the expression of miR-32 was detected in recombinant cell lines by qRT-PCR, and the expression of PTEN and EMT related proteins were detected by Western blot. the ability of metastasis of cervical cancer cells before and after transfection were detected by Transwell. And the relationship were analysis among the expression of miR-32 and PTEN and EMT.Results:(1)miR-32 bind to PTEN with the best specificity and the strongest stability.(2)miR-32 is high expression in cervical cancer tissues and low expression in carcinoma adjacent tissues. PTEN is low expression in cervical cancer tissues and high expression in carcinoma adjacent tissues. These differences were significant(P<0.05).(3)The expression level of miR-32 in poorly differentiated cervical cancer was higher than that in moderate and high differentiated cervical cancer. The expression level of miR-32 in patients of cervical cancer with Lymph node metastasis and distant metastasis were higher than that in patients of cervical cancer without metastasis(P<0.05).(4)miR-32 downregulate in Hela-miR-32 inhibitors cell line and resulting in increasing of PTEN expression level, upregulation of E-cadherin and downregulation of N-cadherin. miR-32 upregulate in Hela-miR-32 mimics cell line and resulting in decreasing of PTEN expression level, downregulation of E-cadherin and upregulation of N-cadherin.(5)The invasion cell count were significantly lower in Hela-miR-32 inhibitors Hela cells than that of Hela, Hela-miR-NC. The invasion cell count were significantly higher in Hela-miR-32 mimics Hela cells than that of Hela, Hela-miR-NC(P<0.05).Conclusions:(1)miR-32 was high expression in cervical cancer and may have relationship to progress and development of cervical cancer.(2) mi RNA-32 can down-regulate PTEN expression and promote the invasion and metastasis of cervical cancer Hela cells.(3) The molecular mechanism of miRNA-32 targeting PTEN for invasion and metastasis of cervical cancer may be related to the down-regulation of E-cadherin and up-regulation of N-cadherin that promote EMT.