Effects of microRNA-221/222 on cell proliferation and apoptosis in prostate cancer cells.
Effects of microRNA-221/222 on cell proliferation and apoptosis in prostate cancer cells.
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microRNA-221/222 对前列腺癌细胞增殖和凋亡的影响。
DOI：
10.1016/j.gene.2015.07.017
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发表时间：
2015-11
期刊：
影响因子：
3.5
通讯作者：
Wang L, Liu C, Li C, Xue J, Zhao S, Zhan P, Jiang A
中科院分区：
文献类型：
--
作者：
Wang L, Liu C, Li C, Xue J, Zhao S, Zhan P, Jiang A
目的探讨miR-221/222在人前列腺癌细胞增殖和凋亡中的作用，并检测miR-221/222对caspase-10表达的影响。方法用miR-221/222模拟物或抑制物转染前列腺癌细胞。MTT法检测细胞增殖情况。采用实时荧光定量PCR检测miR-221/222的表达水平。TNF-α/CHX诱导细胞凋亡，采用Hoechst 33342染色、碘化丙啶（PI）流式细胞术、caspase-3活性测定和Western blot分析。采用荧光素酶活性测定、实时荧光定量PCR和Western blot检测miR-221/222对caspase-10表达的影响。结果miR-221/222能够促进前列腺癌细胞LNCaP和PC3细胞的增殖。转染和诱导凋亡后，Hoechst 33342染色和PI流式细胞术检测显示，miR-221/222模拟物处理的前列腺癌细胞凋亡明显减少。此外，在miR-221/222模拟处理组中，caspase-3活性显著降低，caspase-3的切割形式减少。相反，miR-221/222敲低使前列腺癌细胞对TNF-α/ chx诱导的凋亡敏感。此外，前列腺癌细胞中miR-221/222与caspase-10的表达呈负相关。miR-221/222可以抑制caspase-10的表达，荧光素酶报告基因实验证实了这一点。结论mir -221/222在前列腺癌细胞中通过抑制caspase-10促进细胞增殖，抑制细胞凋亡。我们的研究结果为基于mirna的前列腺癌治疗策略提供了有希望的证据。
ObjectiveTo 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.MethodsProstate 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.ResultsOur 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.ConclusionmiR-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.
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影响因子：
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