肺癌上皮间质转化后引发Nrf2介导的多药耐药的研究

Nrf2-mediated Multidrug Resistance after Epithelial-mesenchymal Transition in Lung Cancer Cells

目的探讨肺癌中上皮间质转化通过Nrf2通路导致多药耐药的可能性。方法 TGF-β1诱导肺腺癌细胞发生上皮间质转化,Western blot法检测癌细胞发生上皮间质转化的标志分子和细胞核内Nrf2的表达变化;比色法测定GSH含量及GST活力的变化;MTT实验检测癌细胞发生上皮间质转化前后对顺铂耐药指数的变化。结果 TGF-β1使癌细胞发生了上皮间质转化:细胞失去极性,上皮标志分子E-cadherin表达下降,而间质标志分子Vimentin和Snail表达上升;细胞发生上皮间质转化后细胞核内Nrf2表达显著增加,GSH含量增高及GST活力增强,癌细胞对顺铂的耐药指数是原来的7.7倍。结论肺癌上皮间质转化后可能通过Nrf2通路介导了癌细胞的多药耐药。

Objective To investigate that possibility of multidrug resistance in lung cancer during Epithelialmesenchymal transition(EMT) through the Nrf2 signaling pahthway. Methods TGF-β1(10ng/ml) was used to induce EMT. The expression of EMT marker genes and nuclear Nrf2 were detected by Western blot. The level of GSH and the activity of GST were measured by colorimetry. MTT assay was used to measure the change of multidrug resistance during EMT. Results TGF-β1(10ng/ml) successfully induced lung adenocarcinoma cells A549 to mesenchymal transition: loss of cell polarity, the decreased expression of epithelial marker E-cadherin, but the increased expression of interstitial marker genes Snail and Vimentin. After the treatment of TGF-β1, the nuclear Nrf2 expression was significantly increased, simultaneously, the level of GSH and the activity of GST were profoundly increased. MTT displayed that the relative multidrug resistance to cisplatin in lung cancer cells was increased by 7.7 times. Conclusion During EMT, lung cancer cells show multidrug resistance, partially through Nrf2 signaling pathway.