Toxic Effects of NNK on NCTC 1469 Cells

doi:10.13560/j.cnki.biotech.bull.1985.2015.09.031

Abstract

Abstract: This work is to investigate the cytotoxicity in NCTC 1469 cells induced by 4-(methyInitrosamino)-1-(3-pyridyl)-1-butanone(NNK). MTT assay was used to analyze the toxic effects on NCTC 1469 cells under various concentrations(0.1, 0.2, 0.3, 0.4, and 0.5 mg/mL)of NNK at different times. Morphologic changes of treated NCTC 1469 cells with NNK were observed by the inverted microscopy. Flow cytometry(FCM)and real-time quantitative PCR were applied to measure the apoptosis rate and the expression of apoptotic gene Bcl-2 and Bax mRNA. NNK depressed the viability of NCTC 1469 cells with a pattern of the dose-dependent and time-dependent. Typical morphological changes of cells shrinkage and the decrease of density were observed by the inverted microscopy. FCW revealed that inhibited cell proliferation and induced death were of dose-dependent, i.e., apoptotic rates were(16.79 ± 2.01)%, (26.87 ± 1.67)%, (41.78 ± 3.19)%, (50.89 ± 3.94)% and(65.86 ± 4.54)% respectively at 24 h after the cells treated with the concentrations(0.1, 0.2, 0.3, 0.4, and 0.5 mg/mL)of NNK, showing much higher than the control of(7.46 ± 1.58)%. Real-time quantitative PCR showed that with the raising of NNH concentration, the expression of gene Bcl-2 decreased and the expression of gene Bax gradually increased. Conclusively, NNK could obviously abate the viability of NCTC 1469 cells and induce the apoptosis of NCTC 1469 cells in a pattern of dose-dependent and time-dependent.

Key words: NNK, NCTC 1469 cells, apoptosis, flow cytometry

Han Yawei, Wang Xihua, Chen Liping, Shi Guiqin, Sun Liping, Zhou Wenshan. Toxic Effects of NNK on NCTC 1469 Cells[J]. Biotechnology Bulletin, 2015, 31(9): 218-223.

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