蝎毒重组蛋白对人肺腺癌A549细胞的抑制作用及其机制  

Inhibitory effect and mechanism of recombinant protein of Buthus martensii venom on human lung adenocarcinoma cell line A549

作　　者：赵灿国[1] 董伟华[1] 孔天翰[2] 

机构地区：[1]广州医学院病理生理学教研室,510182 [2]广州医学院蛇毒研究所,510182

出　　处：《中华生物医学工程杂志》2008年第6期409-412,共4页Chinese Journal of Biomedical Engineering

基　　金：广东省科技计划项目（20088030301370）;广州医学院科研基金项目（2006ZR018）

摘　　要：目的探讨蝎毒重组蛋白对人肺腺癌A549细胞的抑制作用及其抗肿瘤机制。方法四甲基偶氮唑蓝（MTT）比色法检测蝎毒重组蛋白对人肺癌A549细胞增殖的抑制作用。荧光显微镜及流式细胞仪（FCM）技术观测蝎毒重组蛋白对肿瘤细胞内钙离子浓度（[Ca^2＋]i）的影响，从而了解Ca^2＋的可能来源。结果蝎毒重组蛋白对人肺癌A549细胞生长有明显抑制作用（P〈0．05），各浓度组抑制率分别为（25．61±11．0HD）％、（40．91±7．32）％、（65．35±14．51）％。荧光显微镜显示，蝎毒重组蛋白作用细胞48h后细胞内钙离子荧光强度增强，提示[Ca^2＋]i升高。流式细胞仪显示蝎毒重组蛋白在不同情况下可显著增加肿瘤[Ca^2＋]i（P〈0．05）。结论蝎毒重组蛋白对人肺腺癌A549细胞增殖具有显著抑制作用，可能与升高肿瘤[Ca^2＋]i，继而诱导肿瘤细胞凋亡有关；其升高肿瘤[Ca^2＋]i是通过开放肿瘤细胞膜钙通道和肿瘤细胞内钙库释放两条途径实现。Objective To investigate the inhibitory effect and antitumor mechanism of recombinant protein of Buthus martensii venom on human lung adenocarcinoma cell line A549. Methods The inhibition of proliferation in vitro was measured by methyl thiazolyl tetrazolium （MTF） assay. Influence of recombinant protein of Buthus martensii venom on [ Ca2＋ ] i was observed by fluorescence microscope and flow cytometry, in order to explore the source of Ca^2＋ when [Ca^2＋ ] i was alterating. Results The protein of Buthus martensii venom could inhibit A549 cell proliferation（P〈0.05）, the inhihiton rates of different concertration were （25.61 ±11.00）%, （40.91±7.32）% and （65.35± 14.51 ）%. Fluorescent microscope assay showed enhanced fluorescent intensity after 48 hours, which meant the increasing [ Ca^2＋ ] i concentration. Flow cytometry assay showed the recombinant protein of Buthus martensii venom significantly increased the concentration of [ Ca^2＋ ] i in tumor cells under different circumstances（P〈0.05）. Conclusion The recombinant protein of Buthus martensii venom can obviously inhibit the proliferation of A549 cell line, and its mechanism is associated with tumor cell apoptosis through increasing [ Ca^2＋ ] i level of tumor cell as a result of openning the calcium ion channel on the membrane and releasing calcium ion in tumor cells.

关 键 词：重组蛋白质类 蝎毒抗癌多肽 肺肿瘤 A549细胞 抗肿瘤 细胞增殖 细胞凋亡 钙离子 

分 类 号：R285[医药卫生—中药学]