Lee Hwa Young, Kim In Kyoung, Lee Hye In, Mo Jin Young, Yeo Chang Dong, Kang Hyeon Hui, Moon Hwa Sik, Lee Sang Haak
a Division of Respiratory, Allergy and Critical Care Medicine , Department of Internal Medicine, College of Medicine, Seoul St. Mary's Hospital, The Catholic University of Korea , Seoul , Republic of Korea.
b Department of Biochemistry , College of Medicine, The Catholic University of Korea , Seoul , Republic of Korea.
Exp Lung Res. 2016;42(1):14-23. doi: 10.3109/01902148.2015.1125970. Epub 2016 Jan 12.
Statins are known to have pleiotropic effects that induce cell death in certain cancer cells. BIM is a member of the bcl-2 gene family, which promotes apoptotic cell death. This study investigated the hypothesis that simvastatin has pro-apoptotic effects in epidermal growth factor receptor (EGFR)-mutated lung cancer cell lines via the upregulation of the expression of the BIM protein.
The cytotoxic effects of simvastatin on gefitinib-sensitive (HCC827, E716-A750del) and -resistant (H1975, T790M + L858R) nonsmall cell lung cancer (NSCLC) cells were compared. Cell proliferation and expression of apoptosis-related and EGFR downstream signaling proteins were evaluated. Expression of BIM was compared in H1975 cells after treatment with simvastatin or gefitinib. SiRNA-mediated BIM depletion was performed to confirm whether the cytotoxicity of simvastatin was mediated by the expression of BIM.
H1975 cells showed significantly reduced viability compared with HCC827 cells after treatment with simvastatin (2 μM) for 48 hours. In simvastatin-treated H1975 cells, expression of pro-apoptotic proteins was increased and the phosphorylation of ERK 1/2 (p-ERK 1/2) was reduced. Expression of BIM was suppressed by gefitinib (1 μM) treatment in H1975 cells, but it was significantly increased by treatment with simvastatin. BIM depletion by siRNA transfection enhanced the viability of H1975 cells that received simvastatin treatment and increased their expression of anti-apoptotic proteins.
Simvastatin restored the expression of BIM to induce apoptotic cell death in NSCLC cells harboring an EGFR-resistant mutation. Our study suggests the potential utility of simvastatin as a BIM-targeted treatment for NSCLC.
已知他汀类药物具有多效性作用，可诱导某些癌细胞死亡。BIM是bcl-2基因家族的成员，可促进凋亡性细胞死亡。本研究调查了辛伐他汀通过上调BIM蛋白表达对表皮生长因子受体（EGFR）突变的肺癌细胞系具有促凋亡作用这一假说。
比较辛伐他汀对吉非替尼敏感（HCC827、E716 - A750del）和耐药（H1975、T790M + L858R）的非小细胞肺癌（NSCLC）细胞的细胞毒性作用。评估细胞增殖以及凋亡相关蛋白和EGFR下游信号蛋白的表达。比较辛伐他汀或吉非替尼处理后H1975细胞中BIM的表达。进行小干扰RNA（siRNA）介导的BIM缺失实验，以确认辛伐他汀的细胞毒性是否由BIM表达介导。
用2μM辛伐他汀处理48小时后，与HCC827细胞相比，H1975细胞的活力显著降低。在经辛伐他汀处理的H1975细胞中，促凋亡蛋白的表达增加，细胞外信号调节激酶1/2（ERK 1/2）的磷酸化水平降低。在H1975细胞中，1μM吉非替尼处理可抑制BIM的表达，但辛伐他汀处理可使其显著增加。通过siRNA转染使BIM缺失可增强接受辛伐他汀处理的H1975细胞的活力，并增加其抗凋亡蛋白的表达。
辛伐他汀恢复BIM的表达，以诱导携带EGFR耐药突变的NSCLC细胞发生凋亡性细胞死亡。我们的研究表明辛伐他汀作为针对BIM的NSCLC治疗药物具有潜在应用价值。
