Knockdown of c-Met enhances sensitivity to bortezomib in human multiple myeloma U266 cells via inhibiting Akt/mTOR activity
Knockdown of c-Met enhances sensitivity to bortezomib in human multiple myeloma U266 cells via inhibiting Akt/mTOR activity
复制标题
敲低 c-Met 通过抑制 Akt/mTOR 活性增强人多发性骨髓瘤 U266 细胞对硼替佐米的敏感性
DOI：
10.1111/j.1600-0463.2011.02836.x
复制
发表时间：
2012-03-01
期刊：
影响因子：
2.8
通讯作者：
Jiang, Danrong
中科院分区：
文献类型：
--
作者：
Que, Wenzhong;Chen, Junmin;Jiang, Danrong
c-Met是一种受体酪氨酸激酶，在人骨髓瘤细胞系中过表达，并促进骨髓瘤细胞的存活和耐药性。本研究旨在阐明c-Met导致骨髓瘤化疗耐药的机制。使用其中c-Met被有效敲低的稳定U266细胞系并用硼替佐米处理。MTT法检测细胞毒性。流式细胞仪检测细胞周期和凋亡情况。Western blot检测细胞周期相关蛋白的表达、caspase和Akt / mTOR的活性。U266细胞中c-Met敲低降低硼替佐米的平均IC 50，诱导G 0/G1期阻滞，并增加暴露于硼替佐米的U266细胞中caspase介导的凋亡。此外，c-Met敲低降低了细胞周期蛋白D1的水平，并增加了p27和切割的caspase 3和caspase 9的水平。此外，硼替佐米处理的U266细胞中Akt / mTOR活性在c-Met敲低后下调，并且c-Met敲低的U266细胞在Akt和mTOR过表达后恢复化疗抗性。我们的数据表明，c-Met是多发性骨髓瘤的潜在治疗靶点，Akt / mTOR是c-Met保护多发性骨髓瘤细胞免受化疗诱导的生长抑制和凋亡的关键信号组分。
The c-Met is a receptor tyrosine kinase that is overexpressed in human myeloma cell lines and promotes the survival and drug resistance of myeloma cells. This study aimed to elucidate the mechanisms by which c-Met contributes to the chemoresistance in myeloma. Stable U266 cell line in which c-Met was effectively knockdown was employed and treated with bortezomib. Cytotoxicity was evaluated by MTT assay. Cell cycle profile and apoptosis were examined by cytometry analysis. The expression of cell cycle related proteins, and the activities of caspases and Akt / mTOR were detected by Western blot analysis. The c-Met knockdown in U266 cells decreased the average IC50 of bortezomib, induced G0/G1 phase arrest, and increased caspase-mediated apoptosis in U266 cells exposed to bortezomib. In addition, c-Met knockdown decreased the level of cyclin D1 and increased the levels of p27 and cleaved caspase 3 and caspase 9. Moreover, the Akt / mTOR activity in U266 cells treated with bortezomib was downregulated upon c-Met knockdown and c-Met knockdown U266 cells recovered chemoresistance upon the overexpression of Akt and mTOR. Our data demonstrate that c-Met is a potential therapeutic target for multiple myeloma, and Akt / mTOR is a key signaling component through which c-Met protects multiple myeloma cells from chemotherapy-induced growth inhibition and apoptosis.