title: miR-99 inhibits cervical carcinoma cell proliferation by targeting TRIB2.
abstract: MicroRNAs (miRNAs) have significant roles in cell processes, including proliferation, apoptosis and stress responses. To investigate the involvement of miR-99 in the inhibition of HeLa cell proliferation, an miR-99 gene expression vector (pU6.1/miR-99), which overexpressed miR-99 in HeLa cells after transient transfection, was constructed. The expression of miR-99 was detected by qPCR. Cell proliferation and apoptosis were analyzed by cell viability, proliferation and apoptosis assays, as well as by electron microscopy. The results showed that overexpression of miR-99 in HeLa cells increased the HeLa cell mortality rate. Moreover, miR-99 overexpression was able to markedly inhibit HeLa cell proliferation according to the 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The cell apoptosis rate was significantly higher in pU6.1/miR-99-treated cells compared with that in the control cultures. Increases in intracellular electron density, as well as the proportion of nuclear plasma, blebbing phenomena and apoptotic bodies were observed in pU6.1/miR-99-treated cells compared with control cultures according to electron microscopy analysis. The Tribbles 2 (TRIB2) 3'-untranslated region was also observed to be targeted by miR-99 and the results further demonstrated that miR-99 was able to negatively regulate TRIB2 expression in HeLa cells The results indicate that miR-99 acts as a tumor suppressor gene in HeLa cells, establishing a theoretical basis for its application in cancer therapeutics.
authors.affiliation: Key Laboratory of Tumour Molecular Biology in Binzhou Medical University, Department of Biochemistry and Molecular Biology, Binzhou Medical University, Yantai, Shandong 264003, P.R. China.
authors.forename: Jia-Xuan
authors.initials: JX
authors.lastname: Xin
authors.forename: Zhen
authors.initials: Z
authors.lastname: Yue
authors.forename: Shuai
authors.initials: S
authors.lastname: Zhang
authors.forename: Zhong-Hua
authors.initials: ZH
authors.lastname: Jiang
authors.forename: Ping-Yu
authors.initials: PY
authors.lastname: Wang
authors.forename: You-Jie
authors.initials: YJ
authors.lastname: Li
authors.forename: Min
authors.initials: M
authors.lastname: Pang
authors.forename: Shu-Yang
authors.initials: SY
authors.lastname: Xie
keywords: cell proliferation; cervical carcinoma; gene expression; miR-99; microRNAs