Study of molecular mechanism of regulation in cervical cancer progression by long non-coding RNA PTENP1 via miR-3611/PTEN gene pathway
Objective To investigate the molecular mechanism of long non-coding RNA PTENP1(hereinafter referred to as"PTENP1")in the proliferation,migration,and invasion of cervical cancer cells.Methods Cancer tissues and adjacent tissues of 54 patients with cervical cancer diagnosed in Hainan Women and Children's Medical Center from January 2019 to December 2022 were selected,and cervical cancer cell lines(HeLa,SiHa,C33A,Caski)and normal cervical epithelial cell line(H8)were selected.The levels of PTENP1,miR-3611,PTEN gene,and PTEN protein were detected by real-time fluorescent quantitative PCR or Western blot.Appropriate cervical cancer cell lines were selected to compare the expression of PTENP1 in cytoplasm and nucleus,and further divided into blank group(no treatment),pcDNA3.1 group(transfected with pcDNA3.1),pcDNA3.1-PTENP1 group(transfected with pcDNA3.1-PTENP1),NC group(negative control,transfected with eilther mimic or inhibitor,negative control),miR-3611 inhibitor group(transfected with miR-3611 inhibitor),pcDNA3.1-PTENP1+NC group(transfected with pcDNA3.1-PTENP1 and NC),and pcDNA3.1-PTENP1+miR-3611 mimics group(transfected with pcDNA3.1-PTENP1 and miR-3611 mimics).The levels of PTENP1,miR-3611,PTEN gene,PTEN protein,and epithelial mesenchymal transformation related indexes in blank group,pcDNA3.1 group,and pcDNA3.1-PTENP1 group were compared.Cell viability and flow cytometry were used to detect cell proliferation and apoptosis.The levels of miR-3611,PTEN gene,and PTENP1 in blank group,NC group,and miR-3611 inhibitor group were compared.The cell proliferation and epithelial mesenchymal transformation related indexes were compared between pcDNA3.1-PTENP1+NC group and pcDNA3.1-PTENP1+miR-3611 mimics group.Double luciferase reporter gene and RNA pull-down experiments were used to verify the targeting relationship between miR-3611 and PTENP1 and PTEN gene.Results The levels of PTENP1,PTEN gene,and PTEN protein in cancer tissues were lower than those in adjacent tissues,and the levels of miR-3611 were higher than those in adjacent tissues(P<0.05).The levels of PTENP1,PTEN gene,and PTEN protein in HeLa,SiHa,C33A,and Caski cells were lower than those in H8 cells,and the levels of miR-3611 was higher than those in H8 cells(P<0.05).HeLa and Caski cells were selected for subsequent experiments.PTENP1 was highly expressed in HeLa and Caski cytoplasm.The levels of PTENP1,apoptosis rate,E-cadherin,and PTEN gene in pcDNA3.1-PTENP1 group were higher than those in blank group,while cell proliferation activity(cultured 48,72,96 h),and levels of ZEB1,Snail,vimentin,miR-3611 were lower than those in blank group(P<0.05).The levels of miR-3611 in miR-3611 inhibitor group was lower than that in blank group,and PTEN gene and PTENP1 were higher than those in blank group(P<0.05).Cell proliferation activity(cultured 48,72,96 h)and the levels of E-cadherin in pcDNA3.1-PTENP1+miR-3611 mimics group were lower than those in pcDNA3.1-PTENP1+NC group,and the levels of ZEB1,Snail,and vimentin were higher than those in pcDNA3.1-PTENP1+NC group(P<0.05).The PTENP1 levels of HeLa and Caski cells transfected with wild type miR-3611 biotin marker were higher than those of blank cells transfected with biotin marker and cells transfected with mutant type miR-3611 biotin marker,the relative fluorescence activity of 293T cells transfected with PTENP1 or PTEN wild type and miR-3611 mimics was lower than that of 293T cells transfected with PTENP1 or PTEN wild type,respectively(P<0.05).Conclusion PTENP1 regulates the expression of PTEN by competitively binding to miR-3611 and affects the proliferation,migration and invasion of cervical cancer cells.
Long non-coding RNAMicroRNACompetitive binding mechanismPTEN geneCervical cancer