Department of Nephrology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
Int J Biol Markers. 2013 Jun 28;28(2):201-7. doi: 10.5301/JBM.2013.10831.
To identify microRNAs (miRNAs) that are overexpressed in renal cell carcinoma (RCC) and characterize the functional role of miR-21.
The miRNA expression profiles between RCC tissue and adjacent normal tissue were compared using microarray analysis. The differential expression of miR-21 was validated by real-time polymerase chain reaction (RT-PCR). 786-O RCC cells were transfected with miR-21 mimic, miR-21 inhibitor, or negative controls and cell proliferation, apoptosis and cell cycle were examined by MTT assay and flow cytometry. The expression of programmed cell death 4 (PDCD4) and tropomyosin 1 (TPM1) was detected by RT-PCR and Western blot analysis.
Compared to adjacent normal tissue, 10 human miRNAs were significantly upregulated and 7 were downregulated in RCC tissue. RT-PCR confirmed that miR-21 was significantly overexpressed in RCC tissue. In vitro expression of miR-21 mimic promoted the growth of 786-O cells, whereas miR-21 inhibitor inhibited cell growth by inducing apoptosis and cell cycle arrest at S phase. Furthermore, miR-21 mimic or inhibitor significantly reduced or increased the expression of PDCD4 and TPM1.
MiR-21 is overexpressed in RCC tissue and modulates the growth, apoptosis and cell cycle progression of RCC cells and regulates the expression of PDCD4 and TPM1.
鉴定在肾细胞癌（RCC）中过度表达的 microRNAs（miRNAs），并表征 miR-21 的功能作用。
通过微阵列分析比较 RCC 组织和相邻正常组织之间的 miRNA 表达谱。通过实时聚合酶链反应（RT-PCR）验证 miR-21 的差异表达。用 miR-21 模拟物、miR-21 抑制剂或阴性对照转染 786-O RCC 细胞，并通过 MTT 测定和流式细胞术检查细胞增殖、凋亡和细胞周期。通过 RT-PCR 和 Western blot 分析检测程序性细胞死亡 4（PDCD4）和原肌球蛋白 1（TPM1）的表达。
与相邻正常组织相比，RCC 组织中 10 个人类 miRNA 显著上调，7 个 miRNA 下调。RT-PCR 证实 miR-21 在 RCC 组织中显著过表达。体外表达 miR-21 模拟物促进 786-O 细胞的生长，而 miR-21 抑制剂通过诱导细胞凋亡和细胞周期停滞在 S 期抑制细胞生长。此外，miR-21 模拟物或抑制剂显著降低或增加 PDCD4 和 TPM1 的表达。
miR-21 在 RCC 组织中过度表达，调节 RCC 细胞的生长、凋亡和细胞周期进程，并调节 PDCD4 和 TPM1 的表达。
