Crotoxin induces apoptosis and autophagy in human lung carcinoma cells in vitro via activation of the p38 MAPK signaling pathway
Crotoxin induces apoptosis and autophagy in human lung carcinoma cells in vitro via activation of the p38 MAPK signaling pathway
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DOI：
10.1038/aps.2014.62
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发表时间：
2014-10-01
影响因子：
8.2
通讯作者：
Liu, Chun-yu
中科院分区：
文献类型：
--
作者：
Han, Rong;Liang, Hui;Liu, Chun-yu
目的：响尾蛇毒素（CrTX）是南美响尾蛇（Crotalus durissus terrificus）毒液中的主要毒素，在体内外表现出抗肿瘤等药理作用。在此，我们在体外研究了CrTX对人肺癌细胞抗肿瘤作用的分子机制。方法：检测人肺鳞癌SK-MES-1细胞。 CrTX 的细胞毒性在 MIT 和集落形成测定中进行了评估。用流式细胞术研究细胞周期。使用 Hoechst 33258 和膜联蛋白 V-FITC 染色研究细胞凋亡。 Western blot检测相关蛋白水平。结果：CrTX（25、50、100 μmol/L）对SK-MES-1细胞的生长和集落形成有抑制作用，且呈剂量和时间依赖性。 CrTX增加S期细胞的比例并剂量依赖性诱导细胞凋亡，同时下调增殖细胞核抗原（PCNA）的表达，并增加cleaved caspase-3的水平。此外，CrTX剂量依赖性地增加了自噬相关蛋白LC3-II和beclin 1的表达，并降低了细胞中p62的水平。此外，CrTX（50μmol/L）显着增加细胞中p38MAPK的磷酸化。 p38MAPK特异性抑制剂SB203580预处理细胞，可阻断CrTX对细胞增殖的抑制作用，并阻断CrTX诱导的细胞凋亡和裂解caspase-3的表达。结论：p38MAPK信号通路在体外介导CrTX诱导的人肺癌SK-MES-1细胞的凋亡和自噬。
Aim: Crotoxin (CrTX) is the primary toxin in South American rattlesnake (Crotalus durissus terrificus) venom, and exhibits antitumor and other pharmacological actions in vivo and in vitro. Here, we investigated the molecular mechanisms of the antitumor action of CrTX in human lung carcinoma cells in vitro.Methods: Human lung squamous carcinoma SK-MES-1 cells were tested. The cytotoxicity of CrTX was evaluated in both MIT and colony formation assays. Cell cycle was investigated with flow cytometry. Cell apoptosis was studied with Hoechst 33258 and Annexin V-FITC staining. The levels of relevant proteins were analyzed using Western blot assays.Results: CrTX (25, 50, 100 mu mol/L) inhibited the growth and colony formation of SK-MES-1 cells in dose- and time-dependent manners. CrTX increased the proportion of S phase cells and dose-dependently induced cell apoqtosis, accompanied by down-regulating the expression of proliferating cell nuclear antigen (PCNA), and increasing the level of cleaved caspase-3. Furthermore, CrTX dose-dependently increased the expression of autophagy-related proteins LC3-II and beclin 1, and decreased the level of p62 in the cells. Moreover, CrTX (50 mu mol/L) significantly increased p38MAPK phosphorylation in the cells. Pretreatment of the cells with SB203580, a specific inhibitor of p38MAPK, blocked the inhibition of CrTX on cell proliferation, as well as CrTX-induced apoptosis and cleaved caspase-3 expression.Conclusion: The p38MAPK signaling pathway mediates CrTX-induced apoptosis and autophagy of human lung carcinoma SK-MES-1 cells in vitro.