Lina Wang1, Chunyan Liu2, Chaoyang Li2, Jing Xue3, Shihu Zhao2, Panpan Zhan2, Yani Lin2, Pengju Zhang2, Anli Jiang2, Weiwen Chen4. 1. Department of Biochemistry and Molecular Biology, Medical School of Shandong University, Jinan 250012, Shandong, China; Jinan Infectious Disease Hospital, Jinan 250012, Shandong, China. 2. Department of Biochemistry and Molecular Biology, Medical School of Shandong University, Jinan 250012, Shandong, China. 3. Medical School of Shandong University, Jinan 250012, Shandong, China. 4. Department of Biochemistry and Molecular Biology, Medical School of Shandong University, Jinan 250012, Shandong, China. Electronic address: chenweiwen@sdu.edu.cn.
Abstract
OBJECTIVE: To investigate the role of miR-221/222 in cell proliferation and apoptosis in human prostate cancer cells, and examine the effects of miR-221/222 on caspase-10 expression. METHODS: Prostate cancer cells were transfected with miR-221/222 mimics or inhibitors. Cell proliferation was assessed by MTT assay. The expression levels of miR-221/222 were detected with quantitative real-time PCR. Apoptosis was induced with TNF-α/CHX treatment, and evaluated by Hoechst 33342 staining, propidium iodide (PI) flow cytometric analysis, caspase-3 activity measurement, and Western blot analysis. Luciferase activity assay, quantitative real-time PCR, and Western blot were performed to evaluate the effects of miR-221/222 on caspase-10 expression. RESULTS: Our results showed that miR-221/222 could promote the proliferation of prostate cancer cells, including LNCaP and PC3 cells. After transfection and apoptosis induction, Hoechst 33342 staining and PI flow cytometric assay showed that apoptosis was dramatically decreased in prostate cancer cells treated with miR-221/222 mimics. Moreover, caspase-3 activity was dramatically decreased, and the cleaved forms of caspase-3 were reduced, in the miR-221/222 mimic-treated group. On the contrary, miR-221/222 knockdown sensitized the prostate cancer cells to TNF-α/CHX-induced apoptosis. In addition, a negative correlation was observed between the expressions of miR-221/222 and caspase-10 in prostate cancer cells. miR-221/222 could repress the expression of caspase-10, which was confirmed by the luciferase reporter assay. CONCLUSION: miR-221/222 promote cell proliferation and repress apoptosis, through suppressing caspase-10, in prostate cancer cells. Our results provide promising evidence for the miRNA-based therapeutic strategy of prostate cancers.
OBJECTIVE: To investigate the role of miR-221/222 in cell proliferation and apoptosis in humanprostate cancer cells, and examine the effects of miR-221/222 on caspase-10 expression. METHODS:Prostate cancer cells were transfected with miR-221/222 mimics or inhibitors. Cell proliferation was assessed by MTT assay. The expression levels of miR-221/222 were detected with quantitative real-time PCR. Apoptosis was induced with TNF-α/CHX treatment, and evaluated by Hoechst 33342 staining, propidium iodide (PI) flow cytometric analysis, caspase-3 activity measurement, and Western blot analysis. Luciferase activity assay, quantitative real-time PCR, and Western blot were performed to evaluate the effects of miR-221/222 on caspase-10 expression. RESULTS: Our results showed that miR-221/222 could promote the proliferation of prostate cancer cells, including LNCaP and PC3 cells. After transfection and apoptosis induction, Hoechst 33342 staining and PI flow cytometric assay showed that apoptosis was dramatically decreased in prostate cancer cells treated with miR-221/222 mimics. Moreover, caspase-3 activity was dramatically decreased, and the cleaved forms of caspase-3 were reduced, in the miR-221/222 mimic-treated group. On the contrary, miR-221/222 knockdown sensitized the prostate cancer cells to TNF-α/CHX-induced apoptosis. In addition, a negative correlation was observed between the expressions of miR-221/222 and caspase-10 in prostate cancer cells. miR-221/222 could repress the expression of caspase-10, which was confirmed by the luciferase reporter assay. CONCLUSION:miR-221/222 promote cell proliferation and repress apoptosis, through suppressing caspase-10, in prostate cancer cells. Our results provide promising evidence for the miRNA-based therapeutic strategy of prostate cancers.
Authors: Feng Yu; Katherine A Pillman; Corine T Neilsen; John Toubia; David M Lawrence; Anna Tsykin; Michael P Gantier; David F Callen; Gregory J Goodall; Cameron P Bracken Journal: Nucleic Acids Res Date: 2017-11-02 Impact factor: 16.971