microRNA-21 在介导多西紫杉醇耐药中的作用在雄激素非依赖性前列腺癌细胞 PC3 中。
Involvement of microRNA-21 in mediating chemo-resistance to docetaxel in androgen-independent prostate cancer PC3 cells.
机构信息
Department of Urology, Cancer Hospital, Fudan University, Shanghai, China.
出版信息
Acta Pharmacol Sin. 2010 Jul;31(7):867-73. doi: 10.1038/aps.2010.48. Epub 2010 Jun 28.
AIM
To investigate whether microRNA-21 was involved in mediating the chemoresistance of prostate cancer cells to docetaxel.
METHODS
A microarray technique was used to determine the miRNA profile in docetaxel-resistant PC3 cells. Real-time PCR was used to confirm the array results. miR-21 mimics and inhibitors were synthesized and introduced to cells using Lipofectamine 2000. Cell proliferation was examined with the CCK-8 assay. Luciferase reporter containing PDCD 3'UTR was constructed and the activity was detected by a dual luciferase assay. PDCD4 protein expression was evaluated using Western blot.
RESULTS
A docetaxel-resistant prostate cancer PC3 cell line (PC3R) was established . Using microarrays, miR-21 was found to be up-regulated in PC3R cells. Ectopic expression of miR-21 increased the resistance to docetaxel in PC3 wild type cells. In contrast, silencing of miR-21 in PC3R cells sensitized the cells to docetaxel. The IC(50) values for miR-21-silencing cells and control cells were 28.31 and 35.89 nmol/L, respectively. PDCD4, a direct target gene of miR-21, could mediate chemoresistance to docetaxel in PC3 cells.
CONCLUSION
Our findings suggest that miR-21 contributed to the resistance of PC3 cells to docetaxel, and that targeting miR-21 may offer a promising therapeutic approach in sensitizing prostate cancer to docetaxel treatment.
目的
探讨微小 RNA-21 是否参与介导前列腺癌细胞对多西紫杉醇的耐药性。
方法
采用微阵列技术检测多西紫杉醇耐药 PC3 细胞中的 miRNA 谱。实时 PCR 用于确认数组结果。合成 miR-21 模拟物和抑制剂，并使用 Lipofectamine 2000 将其引入细胞。用 CCK-8 测定法检测细胞增殖。构建包含 PDCD3'UTR 的荧光素酶报告基因，并通过双荧光素酶测定法检测其活性。使用 Western blot 评估 PDCD4 蛋白表达。
结果
建立了耐多西紫杉醇的前列腺癌细胞系（PC3R）。使用微阵列，发现 miR-21 在 PC3R 细胞中上调。外源性表达 miR-21 增加了 PC3 野生型细胞对多西紫杉醇的耐药性。相反，沉默 miR-21 可使 PC3R 细胞对多西紫杉醇敏感。miR-21 沉默细胞和对照细胞的 IC50 值分别为 28.31 和 35.89 nmol/L。PDCD4 是 miR-21 的直接靶基因，可介导 PC3 细胞对多西紫杉醇的耐药性。
结论
我们的研究结果表明，miR-21 有助于 PC3 细胞对多西紫杉醇的耐药性，靶向 miR-21 可能为增敏前列腺癌对多西紫杉醇治疗提供有前景的治疗方法。
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