蜂毒肽诱导人肝癌细胞系SMMC-7721程序性坏死

doi:10.16098/j.issn.0529-1356.2021.01.010

解剖学报 ›› 2021, Vol. 52 ›› Issue (1): 67-72.doi: 10.16098/j.issn.0529-1356.2021.01.010

蜂毒肽诱导人肝癌细胞系SMMC-7721程序性坏死

李亚巍张巍* 李妍侯建成张红朱文赫*

吉林医药学院生物化学教研室，吉林吉林 132013

收稿日期:2019-07-31 修回日期:2019-11-07 出版日期:2021-02-06 发布日期:2021-02-06
通讯作者: 张巍;朱文赫 E-mail:huolizwh@163.com
基金资助:
吉林省科技发展计划项目

Effect of melittin on programmed necrosis of human hepatocellular carcinoma cell line SMMC-7721

LI Ya-wei ZHANG Wei* LI Yan HOU Jian-cheng ZHANG Hong ZHU Wen-he*

Department of Biochemistry, Jilin Medical University, Jilin Jilin 132013, China

Received:2019-07-31 Revised:2019-11-07 Online:2021-02-06 Published:2021-02-06
Contact: ZHANG Wei;ZHU Wen-he E-mail:huolizwh@163.com

摘要/Abstract

摘要：

目的探讨蜂毒肽（MLT）对人肝癌细胞系SMMC-7721的杀伤作用及可能机制。方法采用MTT法检测不同浓度蜂毒肽对人肝癌细胞系SMMC-7721的杀伤及程序性坏死特异性抑制剂-1 (Nec-1)对蜂毒肽杀伤SMMC-7721细胞的抑制作用。Hoechst 33342及PI双染色观察细胞程序性坏死的发生，流式细胞术检测细胞程序性坏死率，透射电子显微镜检测细胞超微结构变化，Western blotting法检测SMMC-7721细胞内受体相互作用蛋白1(RIP1)的表达变化。结果与对照组比较，不同浓度蜂毒肽作用SMMC-7721细胞24 h后，细胞增殖活力明显下降(P<0.05)；细胞染色呈深蓝色和红色的形态；细胞膜完整性被破坏、细胞器肿胀、细胞器膜被破坏、核膜完整性丧失，表明细胞发生坏死；Westen blotting结果显示，SMMC-7721细胞内RIP1表达比例升高。与蜂毒肽给药组比较，Nec-1预处理组细胞增殖活力显著提高，细胞坏死率降低，细胞内RIP1表达下调。结论蜂毒肽通过RIP1介导的程序性坏死途径诱导SMMC-7721细胞死亡。

关键词: 蜂毒肽, 肝癌细胞系SMMC-7721, 程序性坏死特异性抑制剂-1, 受体相互作用蛋白1, 免疫印迹法;, 人

Abstract:

Objective To investigate the lethal effect of melittin on human hepatocellular carcinoma cell line SMMC-7721 and its possible mechanism. Methods MTT assay was used to investigate the killing effect of different concentrations melittin on human hepatocellular carcinoma cells SMMC-7721. Meanwhile, the inhibitory effect of specific programmed necrosis inhibitors necrostain-1(Nec-1) on melittin killing SMMC-7721 cells was detected. Programmed cell necrosis observed by Hoechst 33342 and PI double staining. The necrotosis rate was detected by flow cytometry. Ultrastructural changes of cell was detected by transmission electron microscopy. The expression of receptor interacting protein 1(RIP1) was detected by Western blotting. Results Compared with the control group, the proliferation activity of SMMC-7721 cells was significantly decreased after treatment with different concentrations of melittin for 24 hours (P<0.05). Cells stained in dark blue and red. Cell membrane integrity was destroyed, organelle swelling, organelle membrane was destroy, that demonstrates cell was necrosis. Westen blotting result showed an increased proportion of RIP1 expression in SMMC-7721 cells. Compared with the melittin group, cell proliferation activity was significantly increased, cell necrotosis rate was decreased, and intracellular RIP1 expression was down-regulated in the Nec-1 pretreatment group. Conclusion Melittin induces cell death in SMMC-7721 cells through the RIP1-mediated programmed necrosis pathway.

Key words: Melittin, Hepatoma cell line SMMC-7721, Specific programmed necrosis inhibitors necrostatin-1, Receptor interacting protein 1, Western blotting, Human

中图分类号:

R735.9

李亚巍张巍李妍侯建成张红朱文赫. 蜂毒肽诱导人肝癌细胞系SMMC-7721程序性坏死[J]. 解剖学报, 2021, 52(1): 67-72.

LI Ya-wei ZHANG Wei LI Yan HOU Jian-cheng ZHANG Hong ZHU Wen-he. Effect of melittin on programmed necrosis of human hepatocellular carcinoma cell line SMMC-7721[J]. Acta Anatomica Sinica, 2021, 52(1): 67-72.

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蜂毒肽诱导人肝癌细胞系SMMC-7721程序性坏死

Effect of melittin on programmed necrosis of human hepatocellular carcinoma cell line SMMC-7721

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