Zhiwei Zhong1, Zhuo Dong, Lihua Yang, Zhaohui Gong. 1. Institute of Biochemistry and Molecular Biology, School of Medicine, Ningbo University, 818 Fenghua Road, Ningbo 315211, China.
Abstract
PURPOSE: MicroRNAs regulate critical genes associated with lung cancer. Human mutS homolog 2 (hMSH2), one of the core mismatch repair genes, is affected in lung cancer development. The aim of this study is to investigate the role of miR-21 in hMSH2 gene expression and the effect of miR-21 on cell proliferation and cell cycle in lung cancer. METHODS: The targets of miR-21 were predicted by a bioinformatics tool, and hMSH2 was validated as a direct target of miR-21 by luciferase activity assay. MiRNA mimics or inhibitors were used to stimulate or attenuate the effect of endogenous miR-21 on hMSH2 expression. MiR-21 and hMSH2 expressions were assessed with real-time RT-PCR and Western blotting. Cell cycle was determined by flow cytometry, and cell growth was analyzed by MTT assay and real-time cell analysis system. RESULTS: MiR-21 expression was inversely correlated with hMSH2 expression in human lung cancer cell lines. Further validation showed hMSH2 was directly regulated by miR-21. The up-regulation of miR-21 significantly promoted cell proliferation and revealed a higher proportion of cells at S phase. However, knockdown of miR-21 expression resulted in cell cycle arrest at G2/M phase and inhibited cell proliferation. CONCLUSIONS: These data suggest miR-21 is a key regulator of hMSH2 and modulates cell cycle and proliferation by targeting hMSH2 in human lung cancer.
PURPOSE: MicroRNAs regulate critical genes associated with lung cancer. HumanmutS homolog 2 (hMSH2), one of the core mismatch repair genes, is affected in lung cancer development. The aim of this study is to investigate the role of miR-21 in hMSH2 gene expression and the effect of miR-21 on cell proliferation and cell cycle in lung cancer. METHODS: The targets of miR-21 were predicted by a bioinformatics tool, and hMSH2 was validated as a direct target of miR-21 by luciferase activity assay. MiRNA mimics or inhibitors were used to stimulate or attenuate the effect of endogenous miR-21 on hMSH2 expression. MiR-21 and hMSH2 expressions were assessed with real-time RT-PCR and Western blotting. Cell cycle was determined by flow cytometry, and cell growth was analyzed by MTT assay and real-time cell analysis system. RESULTS:MiR-21 expression was inversely correlated with hMSH2 expression in humanlung cancer cell lines. Further validation showed hMSH2 was directly regulated by miR-21. The up-regulation of miR-21 significantly promoted cell proliferation and revealed a higher proportion of cells at S phase. However, knockdown of miR-21 expression resulted in cell cycle arrest at G2/M phase and inhibited cell proliferation. CONCLUSIONS: These data suggest miR-21 is a key regulator of hMSH2 and modulates cell cycle and proliferation by targeting hMSH2 in humanlung cancer.
Authors: Xi Liu; Lorenzo F Sempere; Haoxu Ouyang; Vincent A Memoli; Angeline S Andrew; Yue Luo; Eugene Demidenko; Murray Korc; Wei Shi; Meir Preis; Konstantin H Dragnev; Hua Li; James Direnzo; Mads Bak; Sarah J Freemantle; Sakari Kauppinen; Ethan Dmitrovsky Journal: J Clin Invest Date: 2010-03-08 Impact factor: 14.808
Authors: Kenji Zennami; Su Mi Choi; Ross Liao; Ying Li; Wikum Dinalankara; Luigi Marchionni; Fatema H Rafiqi; Akira Kurozumi; Koji Hatano; Shawn E Lupold Journal: Mol Cancer Res Date: 2018-12-05 Impact factor: 5.852