Department of Oncology, Binzhou Medical College Affiliated Hospital, 661#, Yellow-River Second Street, 256603 Binzhou, Shandong Province, China.
Diagn Pathol. 2014 Jul 10;9:143. doi: 10.1186/1746-1596-9-143.
Overexpression of microRNA-182 (miR-182) is found in various human cancers, including non-small cell lung cancer (NSCLC). Our aim is to investigate the association of miR-182 expression with the sensitivity of NSCLC to cisplatin.
TaqMan RT-PCR or Western blot assay was performed to detect the expression of mature miR-182 and programmed cell death 4 (PDCD4) protein. miR-182 and (or) PDCD4 depleted cell lines were generated using miR-182 inhibitor and (or) siRNA. The viabilities of treated cells were analyzed using MTT assay.
The expression level of miR-182 in A549 cell line was significantly higher than that in NHBE cell line (p < 0.01). Transfection of miR-182 inhibitor induced sensitivity of A549 cells to cisplatin. A549 cells transfected with PDCD4 siRNA became more resistant to cisplatin therapy. We found an increase PDCD4 protein level following the transfection of miR-182 inhibitor using Western blot analysis. In addition, the enhanced growth-inhibitory effect by miR-182 inhibitor was weakened after the addition of PDCD4 siRNA.
The results of the present study demonstrated that overexpression of miR-182 may involve in chemoresistance of NSCLC cells to cisplatin by down-regulating PDCD4.
The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1793467320130186.
在多种人类癌症中发现 microRNA-182（miR-182）表达过度，包括非小细胞肺癌（NSCLC）。我们的目的是研究 miR-182 表达与 NSCLC 对顺铂敏感性的关系。
采用 TaqMan RT-PCR 或 Western blot 检测成熟 miR-182 和程序性细胞死亡 4（PDCD4）蛋白的表达。采用 miR-182 抑制剂和（或）siRNA 生成 miR-182 和（或）PDCD4 耗竭细胞系。采用 MTT 分析检测处理细胞的活力。
A549 细胞系中 miR-182 的表达水平明显高于 NHBE 细胞系（p<0.01）。转染 miR-182 抑制剂诱导 A549 细胞对顺铂的敏感性。用 PDCD4 siRNA 转染的 A549 细胞对顺铂治疗的耐药性增加。我们发现 Western blot 分析显示，转染 miR-182 抑制剂后 PDCD4 蛋白水平升高。此外，加入 PDCD4 siRNA 后，miR-182 抑制剂增强的生长抑制作用减弱。
本研究结果表明，miR-182 的过表达可能通过下调 PDCD4 参与 NSCLC 细胞对顺铂的化疗耐药。
