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

doi:10.16098/j.issn.0529-1356.2021.01.010

PDF(6413 KB)

Acta Anatomica Sinica ›› 2021, Vol. 52 ›› Issue (1) : 67-72. DOI: 10.16098/j.issn.0529-1356.2021.01.010

Cancer Biology

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*

Author information +

History +

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

Cite this article

EndNote

Ris (Procite)

Bibtex

Download Citations

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 https://doi.org/10.16098/j.issn.0529-1356.2021.01.010

References

［1］ Pinoangeles A, Lazaridis T. Effects of peptide charge, orientation, and concentration on melittin transmembrane pores［J］. Biophys J, 2018, 114(12):2865-2874.

［2］ Terwilliger TC, Eisenberg D. The structure of melittin. Ⅱ. Interpretation of the structure ［J］. J Biol Chem, 1982, 257(11): 6016-6022.

［3］ Zhang SF, Chen Z. Melittin exerts an antitumor effect on non-small cell lung cancer cells ［J］. Mol Med Rep, 2017, 16(3): 3581-3586.

［4］ Rady I, Siddiqui IA, Rady M, et al. Melittin, a major peptide component of bee venom, and its conjugates in cancer therapy ［J］. Cancer Lett, 2017, 402: 16-31.

［5］ Park JH, Park B, Park KK. Suppression of hepatic epithelial-to-mesenchymal transition by melittin via blocking of TGFβ/Smad and MAPK-JNK signaling pathways ［J］. Toxins (Basel), 2017,9(4): 138-150.

［6］ Nault JC. The end of almost 10 years of negative RCTs in advanced hepatocellular carcinoma ［J］. Lancet, 2017, 389(10064): 4-6.

［7］ Xu L, Zhang M, Zheng X, et al. The circular RNA ciRS-7 (Cdr1as) acts as a risk factor of hepatic microvascular invasion in hepatocellular carcinoma ［J］. J Cancer Res Clin Oncol, 2017, 143(1): 17-27.

［8］ Zhang H， Zhang T， Fang YSh， et al． Inhibitory effect of artemisinin combined with cisplatinon on HepG2 cells proliferation ［J］． Journal of Toxicology， 2016，(6): 435-440． (in Chinase)

张红, 张婷, 凡亚帅, 等. 青蒿素联合顺铂抑制肝癌细胞HepG2增殖的影响［J］. 毒理学杂志, 2016, (6): 435-440.

［9］ Iannielli A, Bido S, Folladori L, et al. Pharmacological inhibition of necroptosis protects from dopaminergic neuronal cell death in parkinson’s disease models［J］. Cell Reports, 2018, 22(8):2066-2079.

［10］ Jorgensen I, Rayamajhi M, Miao EA. Programmed cell death as a defence against infection ［J］. Nat Rev Immunol, 2017, 17(3): 151-164.

［11］ Kondylis V, Kumari S, Vlantis K, et al. The interplay of IKK, NF-κB and RIPK1 signaling in the regulation of cell death, tissue homeostasis and inflammation ［J］. Immunol Rev, 2017, 277(1): 113-127.

［12］ Zhu WH, Xu N, Xu JJ, et al. Effects of dihydroartemisinin on proliferation and apoptosisof human pancreatic cancer cell line ［J］. Acta Anatomica Sinica, 2018, 49(1): 70-74. (in Chinase)

朱文赫, 许娜, 徐俊杰, 等. 双氢青蒿素对人胰腺癌细胞系增殖和凋亡的影响［J］. 解剖学报, 2018, 49(1), 70-74.

［13］ Festjens N, Berghe TV, Cornelis S, et al. RIP1, a kinase on the crossroads of a cell’s decision to live or die［J］. Cell Death Differ, 2007, 14(3):400-410.

［14］ Harris PA, Berger SB, Jeong JU, et al. Discovery of a first-in-class receptor interacting protein 1 (RIP1) kinase specific clinical candidate (GSK2982772) for the treatment of inflammatory diseases ［J］. J Med Chem, 2017, 60(4): 1247-1261.

［15］ Wegner KW, Saleh D, Degterev A. Complex pathologic roles of RIPK1 and RIPK3: moving beyond necroptosis ［J］. Trends Pharmacol Sci, 2017, 38(3): 202-225.

［16］ Chen S, Lv X, Hu B, et al. RIPK1/RIPK3/MLKL-mediated necroptosis contributes to compression-induced rat nucleus pulposus cells death ［J］. Apoptosis, 2017, 22(5): 626-638.

［17］ Zhang L, Li R, Jiang R, et al. A comparative study on the effect of four-source ultrasonic power on the microstructure and mechanical properties of large-scale 2219 aluminum ingots［J］. JOM, 2019, 71(6):2063-2071.