Institute for Chemical Carcinogenesis, State Key Laboratory of Respiratory Diseases, Guangzhou Medical University, Guangzhou 510182, PR China.
Toxicol In Vitro. 2010 Apr;24(3):928-35. doi: 10.1016/j.tiv.2009.11.013. Epub 2009 Nov 16.
MicroRNAs (miRNAs) are a class of small noncoding RNA molecules with profound impact on various biological processes. Some miRNAs are involved in tumorigenesis by regulation of cell cycle progression. Here, we cultured primary murine bronchial epithelial cells and then examined the expression of miR-320 and miR-494 in cells exposed to benzo[a]pyrene (B[a]P). To better characterize roles of miR-320 and miR-494 in cell cycle progression, we used miRNA inhibitors to downregulate expression of miRNAs and determined cell cycle distribution and expression of cyclin-dependent kinases 6 (CDK6) by flow cytometric analysis. Treating cells with 1 microM B[a]P for 24h resulted in time-dependent increases in miR-320 and miR-494 expression. Moreover, G1 arrest and downregulated expression of CDK6 were shown in the treated cells. Flow cytometric analysis indicated a relief of G1 arrest and an elevated expression of CDK6 after inhibition of the expressions of miR-320 and miR-494 in cells exposed to B[a]P. These results suggest that expression levels of miRNA-320 and miR-494, which regulate B[a]P-exposed cell cycle progression, may impact G1/S transition through CDK6, and provide further insights into functions of miRNAs in cell cycle of primary murine bronchial epithelial cells exposed to B[a]P.
微小 RNA(miRNAs)是一类具有深远影响的小型非编码 RNA 分子,参与各种生物过程。一些 miRNAs 通过调节细胞周期进程参与肿瘤发生。在这里,我们培养了原代鼠支气管上皮细胞,然后检测了暴露于苯并[a]芘(B[a]P)的细胞中 miR-320 和 miR-494 的表达。为了更好地描述 miR-320 和 miR-494 在细胞周期进程中的作用,我们使用 miRNA 抑制剂下调 miRNA 的表达,并通过流式细胞术分析确定细胞周期分布和细胞周期蛋白依赖性激酶 6(CDK6)的表达。用 1μM B[a]P 处理细胞 24h 导致 miR-320 和 miR-494 的表达随时间增加。此外,在处理的细胞中观察到 G1 期阻滞和 CDK6 表达下调。流式细胞术分析表明,在暴露于 B[a]P 的细胞中抑制 miR-320 和 miR-494 的表达后,G1 期阻滞得到缓解,CDK6 的表达升高。这些结果表明,调节 B[a]P 暴露细胞周期进程的 miR-320 和 miR-494 的表达水平可能通过 CDK6 影响 G1/S 转换,并为 miRNA 在 B[a]P 暴露的原代鼠支气管上皮细胞细胞周期中的功能提供了进一步的见解。