Zhihua Chen1, Mengzhen Zhang1, Yuhuan Qiao1, Junjuan Yang2, Qinan Yin3. 1. a The First Affiliated Hospital of Zhengzhou University , Zhengzhou , China. 2. b Women&infants Hospital Of Zhengzhou , Zhengzhou , China. 3. c National Institutes of Health , Bethesda , MD , USA.
Abstract
BACKGROUND: The human cervical carcinoma oncogenic mechanisms still remain elusive. Thus, we proposed to understand the biological role of a newly discovered therapeutic miRNA. METHODS: MiR-1297 related to human cervical carcinoma was selected for this study. TaqMan qRT- PCR assay was used to profile miRNA, phosphatase and tensin homolog (PTEN) expression in randomly chosen tumour with non-tumour tissues, and the apoptosis factors expression. Cell proliferation was monitored by CCK-8 assay and colony formation assay. Apoptosis was determined by flow cytometry. Protein level was determined by western blotting. 3'UTR was performed to validate the direct binding sites of miR-1297 on PTEN. SPSS was used for statistical analyses. RESULTS: MiR-1297 is repressed and PTEN activated in human cervical cancer tissues. After miR-1297 overexpression, HeLa cells had an increase in cell proliferation and decrease in apoptosis. PTEN expression is negatively correlation with miR-1297. PTEN silencing display the similar pattern as miRNA-1297 overexpression to inhibit HeLa cell growth and apoptosis in vitro. CONCLUSIONS: Our data indicate that miR-1297 contribute to the human cervical carcinoma through PTEN. miR-1297 could be a reasonable miRNA for future studies.
BACKGROUND: The human cervical carcinoma oncogenic mechanisms still remain elusive. Thus, we proposed to understand the biological role of a newly discovered therapeutic miRNA. METHODS:MiR-1297 related to human cervical carcinoma was selected for this study. TaqMan qRT- PCR assay was used to profile miRNA, phosphatase and tensin homolog (PTEN) expression in randomly chosen tumour with non-tumour tissues, and the apoptosis factors expression. Cell proliferation was monitored by CCK-8 assay and colony formation assay. Apoptosis was determined by flow cytometry. Protein level was determined by western blotting. 3'UTR was performed to validate the direct binding sites of miR-1297 on PTEN. SPSS was used for statistical analyses. RESULTS:MiR-1297 is repressed and PTEN activated in human cervical cancer tissues. After miR-1297 overexpression, HeLa cells had an increase in cell proliferation and decrease in apoptosis. PTEN expression is negatively correlation with miR-1297. PTEN silencing display the similar pattern as miRNA-1297 overexpression to inhibit HeLa cell growth and apoptosis in vitro. CONCLUSIONS: Our data indicate that miR-1297 contribute to the human cervical carcinoma through PTEN. miR-1297 could be a reasonable miRNA for future studies.