Acquired Cisplatin Resistance in Human Lung Adenocarcinoma Cells Is Associated with Enhanced Autophagy
Acquired Cisplatin Resistance in Human Lung Adenocarcinoma Cells Is Associated with Enhanced Autophagy
复制标题
人肺腺癌细胞获得性顺铂耐药与自噬增强相关
DOI：
10.1089/cbr.2009.0701
复制
发表时间：
2010-02-01
影响因子：
3.4
通讯作者：
Wu, Gang
中科院分区：
文献类型：
--
作者：
Ren, Jing-Hua;He, Wen-Shan;Wu, Gang
自噬激活是接受化疗的肿瘤细胞的一个标志，但自噬在肺腺癌对顺铂类化疗获得性耐药中的作用尚不清楚。我们的目的是通过检测亲代肺腺癌细胞株A549及其8倍、更耐药的亚细胞系A549/DDP的自噬活性来解决这个问题，A549/DDP是通过增加顺铂浓度而获得的。将A549/DDP和A549细胞分别置于无血清培养液和电离辐射中。为了检测应激诱导的自噬，采用免疫荧光法和免疫印迹法检测自噬标志物LC3-II的表达。为了确定已知的自噬抑制剂3-MA在克服获得性顺铂耐药中的作用，采用了四甲基偶氮唑盐比色法和流式细胞术。Western blotting分析表明，在应激条件下，A549/DDP细胞与亲本A549细胞相比，LC3-II表达增加。免疫荧光染色显示，LC3-II蛋白主要定位于A549/DDP细胞的胞浆中。我们还发现，3-MA可以增强顺铂对获得性耐药细胞(A549/DDP)的生长抑制和凋亡作用。总之，我们的结果提供了证据，证明自噬上调在A549/DDP细胞对顺铂耐药中起主要作用。
Activation of autophagy is a hallmark in tumor cells treated with chemotherapy, but the role of autophagy in acquired resistance of lung adenocarcinoma to cisplatin-based chemotherapy remains to be clarified. Our aim was to address that question by surveying the autophagic activity in parental lung adenocarcinoma cell line A549 and its 8-fold, more resistant subcell line, A549/DDP, which was obtained by treating cisplatin with increasing concentrations. A549/DDP and A549 cells were exposed to serum-free culture medium or ionizing radiation. To measure the stress-induced autophagy, LC3-II, as an autophagosome marker, was measured by immunofluorescence and Western blotting. To determine the effect of 3-MA, a known inhibitor of autophagy, on overcoming acquired cisplatin resistance, the MTT assay and flow cytometry were performed. Western blotting analysis demonstrated that LC3-II was increased in A549/DDP cells, compared with those of parental A549 cells, under stress conditions. Meanwhile, immunofluorescence staining showed that LC3-II protein was located mainly in the cytoplasm of A549/DDP. We also found that 3-MA can enhance the growth inhibition and apoptotic effect of cisplatin in acquired resistant cells (A549/DDP). Collectively, our results provide evidence that the upregulation of autophagy plays a major role in cisplatin resistance of A549/DDP cells.