title: [miR-143 inhibits cell proliferation through targeted regulating the expression of K-ras gene in HeLa cells].
abstract: Objective:To explore the effect of microRNA miR-143 on the proliferation of cervical cancer HeLa cells through targeted regulating the expression of K-ras gene.Methods:The luciferase report carrier containing wild type 3'-UTR of K-ras gene (K-ras-wt) or mutated 3'-UTR of the K-ras (K-ras-mut) were co-transfected with iR-143 mimic into the HeLa cells respectively, and the targeting effect of miR-143 in the transfectants was verified by the dual luciferase report system. HeLa cells were also transfected with miR-143 mimic (miR-143 mimic group), mimic control (negative control group), and miR-143 mimic plus K-ras gene (miR-143 mimic+ K-ras group), respectively. The expression of miR-143 in the transfected HeLa cells was detected by real-time PCR (RT-PCR), and the expression of K-ras protein was detected by Western blot. The cell proliferation activity of each group was examined by MTT assay. In addition, human cervical cancer tissue samples (n=5) and cervical intraepithelial neoplasia tissue samples (n=5) were also examined for the expression of miR-143 and K-ras protein by RT-PCR and Western blot, respectively.Results:The luciferase report assay showed that co-transfection with miR-143 mimic decreased the luciferase activity of the K-ras-wt significantly, but did not inhibit the luciferase activity of the K-ras-mut. The expression of miR-143 in the HeLa cells transfected with miR-143 mimic was significantly higher than that in the HeLa cells transfected with the mimic control (3.31±0.45 vs 0.97±0.22,P<0.05). The MTT assay revealed that the cell proliferative activity of the miR-143 mimic group was significantly lower than that of the negative control group (P<0.05), and the cell proliferative activity of the miR-143 mimic+ K-ras group was also significantly lower than the control group (P<0.05) but higher than the miR-143 mimic group significantly (P<0.05). The expression levels of K-ras protein in the miR-143 mimic group, the negative control group and the miR-143 mimic+ K-ras group were lowest, moderate, and highest, respectively (115.27±34.08, 521.36±41.89, and 706.52±89.44, allP<0.05). In the tissue samples, the miR-143 expression in the cervical cancer group was significantly lower than that of the cervical intraepithelial neoplasia group (0.32±0.06 vs. 0.93±0.17,P<0.05); whereas the K-ras protein expression in the cervical cancer group was significantly higher than that in the cervical intraepithelial neoplasia group (584.39±72.34 vs. 114.23±25.82,P<0.05).Conclusions:In vitro, miR-143 can inhibit the proliferative activity of HeLa cells through targeted regulating the expression of K-ras gene. In human cervical cancer tissues of a small sample set, the expression of miR-143 is downregulated, and the expression of K-ras is upregulated.
authors.affiliation: Department of Obstetrics and Gynecology, The First Affiliated Hospital of Xinxiang Medical University, Weihui 453100, China.
authors.forename: H X
authors.initials: HX
authors.lastname: Qin
authors.affiliation: Department of Interventional Radiology, The First Affiliated Hospital of Xinxiang Medical University, Weihui 453100, China.
authors.forename: H K
authors.initials: HK
authors.lastname: Cui
authors.affiliation: Department of Obstetrics and Gynecology, The Third Affiliated Hospital of Xinxiang Medical University, Xinxiang 453000, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Pan
authors.affiliation: Department of Obstetrics and Gynecology, The First Affiliated Hospital of Xinxiang Medical University, Weihui 453100, China.
authors.forename: R L
authors.initials: RL
authors.lastname: Hu
authors.affiliation: Department of Obstetrics and Gynecology, The First Affiliated Hospital of Xinxiang Medical University, Weihui 453100, China.
authors.forename: L H
authors.initials: LH
authors.lastname: Zhu
authors.affiliation: Department of Obstetrics and Gynecology, The First Affiliated Hospital of Xinxiang Medical University, Weihui 453100, China.
authors.forename: S J
authors.initials: SJ
authors.lastname: Wang
mesh_terms: D049109:Cell Proliferation / Q000235:genetics*; D015536:Down-Regulation; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D017930:Genes, Reporter; D011905:Genes, ras; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics; D035683:MicroRNAs / Q000502:physiology*; D009154:Mutation; D060888:Real-Time Polymerase Chain Reaction; D014162:Transfection; D002583:Uterine Cervical Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D018631:ras Proteins / Q000378:metabolism
chemical_list: C542359:MIRN143 microRNA, human; D035683:MicroRNAs; D008156:Luciferases; D018631:ras Proteins