Study on the mechanism of human large cell lung cancer NCIH661 cells apoptosis induced by crotoxin

doi:10.3760/cma.j.issn.1673-422X.2015.07.001

Abstract

Abstract: ObjectiveTo observe the apoptosis of human large cell lung cancer NCIH661 cells induced by crotoxin, and to explore its mechanism. MethodsThe growth suppression of crotoxin on the NCIH661 cells was detected by CCK8 colorimetry, and the formation of NCIH661 cells was observed by the plat colony experiment. This experiment included 4 groups: negative control group, crotoxin group (60 μg/ml crotoxin acted for 24 h), crotoxin +SB203580 group (pretreated cells using 5 μmol/L SB203580 for 1 h, then 60 μg/ml crotoxin acted for 24 h), SB203580 group (pretreated cells using 5 μmol/L SB203580 for 1 h, then cultivated cells using complete culture solution). They were detected that the cell cycle and apoptosis rate of NCIH661 cells treated with crotoxin by the flow cytometry. Additionally, they were tested that the change of the cell cycle and apoptosis rate after the NCIH661 cells were treated with crotoxin and the activity of p38MAPK was inhibited by SB203580. ResultsWhen the concentration of crotoxin was greater than or equal to 30 μg/ml, the inhibitory effect of crotoxin on the activity of NCIH661 cells and colony formation, and inhibition rate rose with increasing function of time and drug concentration. Flow cytometry showed that the apoptosis rate of crotoxin group and crotoxin+SB203580 group were (16.70±1.38)% and (2.15±0.54)%, compared to the control group (1.47±0.29)%, and the former difference was statistically significant and the latter was not statistically significant(t=-18.763, P=0.000; t=-1.935, P=0.125). The G1 period cells of crotoxin group and crotoxin+SB203580 group were (57.25±1.09)% and (48.04±1.03)%, compared to the control group (47.46±0.69)%, and the former difference was statistically significant and the latter was not statistically significant(t=-13.124, P=0.000; t=-0.809, P=0.464). ConclusionCrotoxin can promote the apoptosis of human large cell lung cancer NCIH661 cells, and this effect may be related to the excitation of p38MAPK signal pathway.

Key words: Neurotoxins, Carcinoma, nonsmallcell lung, Apoptosis, p38MAPK

Li Rui, He Jingkang, Shen Jian, Shi Zhen, Li Wei, Yu Xiaojun. Study on the mechanism of human large cell lung cancer NCIH661 cells apoptosis induced by crotoxin[J]. Journal of International Oncology, 2015, 42(7): 481-484.

References

［1］ Rivera MP. Multimodality therapy in the treatment of lung cancer［J］. Semin Respir Crit Care Med, 2004, 25 Suppl 1: 3-10. ［2］ Favoretto BC, Ricardi R, Silva SR, et al. Immunomodulatory effects of crotoxin isolated from Crotalus durissus terrificus venom in mice immunised with human serum albumin［J］. Toxicon, 2011, 57(4): 600-607. ［3］ Nunes FP, Zychar BC, DellaCasa MS, et al. Crotoxin is responsible for the longlasting antiinflammatory effect of Crotalus durissus terrificus snake venom: involvement of formyl peptide receptors［J］. Toxicon, 2010, 55(6): 1100-1106. ［4］ Yan CH, Yang YP, Qin ZH, et al. Autophagy is involved in cytotoxic effects of crotoxin in human breast cancer cell line MCF7 cells［J］. Acta Pharmacol Sin, 2007, 28(4): 540-548. ［5］ Maddika S, Ande SR, Panigrahi S, et al. Cell survival, cell death and cell cycle pathways are interconnected: implications for cancer therapy［J］. Drug Resist Updat, 2007, 10(12): 13-29. ［6］ Yan CH, Liang ZQ, Gu ZL, et al. Contributions of autophagic and apoptotic mechanisms to CrTXinduced death of K562 cells［J］. Toxicon, 2006, 47(5): 521-530. ［7］ Liu Z, Ma Y, Moyer MP, et al. Involvement of the mannose receptor and p38 mitogenactivated protein kinase signaling pathway of the microdomain of the integral membrane protein after enteropathogenic Escherichia coli infection［J］. Infect Immun, 2012, 80(4): 1343-1350. ［8］ Wu JG, Tang H, Liu ZJ, et al. Angiotensin(17) inhibits vascular remodelling in rat jugular vein grafts via reduced ERK1/2 and p38MAPK activity［J］. J Int Med Res, 2011, 39(6): 2158-2168. ［9］ BretonRomero R, Gonzalez de Orduna C, Romero N, et al. Critical role of hydrogen peroxide signaling in the sequential activation of p38MAPK and eNOS in laminar shear stress［J］. Free Radic Biol Med, 2012, 52(6): 1093-1100. ［10］ Okamura S, Arakawa H, Tanaka T, et al. p53DINP1, a p53inducible gene, regulates p53dependent apoptosis［J］. Mol Cell, 2001, 8(1): 85-94. ［11］ Kerr JF, Winterford CM, Harmon BV. Apoptosis. Its significance in cancer and cancer therapy［J］. Cancer, 1994, 73(8): 2013-2026. ［12］ Tamm I, Schriever F, Dorken B. Apoptosis: implications of basic research for clinical oncology［J］. Lancet Oncol, 2001, 2(1): 33-42. ［13］ Huang X, Halicka HD, Traganos F, et al. Cytometric assessment of DNA damage in relation to cell cycle phase and apoptosis［J］. Cell Prolif, 2005, 38(4): 223-243.

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