Wang Lina, Liu Chunyan, Li Chaoyang, Xue Jing, Zhao Shihu, Zhan Panpan, Lin Yani, Zhang Pengju, Jiang Anli, Chen Weiwen
Department of Biochemistry and Molecular Biology, Medical School of Shandong University, Jinan 250012, Shandong, China; Jinan Infectious Disease Hospital, Jinan 250012, Shandong, China.
Department of Biochemistry and Molecular Biology, Medical School of Shandong University, Jinan 250012, Shandong, China.
Gene. 2015 Nov 10;572(2):252-8. doi: 10.1016/j.gene.2015.07.017. Epub 2015 Jul 9.
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.
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.
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.
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.
研究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。
用miR - 221/222模拟物或抑制剂转染前列腺癌细胞。通过MTT法评估细胞增殖。用定量实时PCR检测miR - 221/222的表达水平。用TNF-α/CHX处理诱导凋亡，并通过Hoechst 33342染色、碘化丙啶（PI）流式细胞术分析、caspase - 3活性测定和蛋白质印迹分析进行评估。进行荧光素酶活性测定、定量实时PCR和蛋白质印迹以评估miR - 221/222对caspase - 10表达的影响。
我们的结果表明，miR - 221/222可促进前列腺癌细胞（包括LNCaP和PC3细胞）的增殖。转染并诱导凋亡后，Hoechst 33342染色和PI流式细胞术检测显示，用miR - 221/222模拟物处理的前列腺癌细胞凋亡显著减少。此外，在miR - 221/222模拟物处理组中，caspase - 3活性显著降低，caspase - 3的裂解形式减少。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，在前列腺癌细胞中观察到miR - 221/222与caspase - 10的表达呈负相关。荧光素酶报告基因测定证实，miR - 221/222可抑制caspase - 10的表达。
miR - 221/222通过抑制caspase - 10促进前列腺癌细胞的增殖并抑制凋亡。我们的结果为基于miRNA的前列腺癌治疗策略提供了有前景的证据。
