[关键词]
[摘要]
该研究探究了呕吐毒素抑制HK2细胞的增殖与诱导凋亡蛋白表达作用。不同浓度的呕吐毒素作用于HK2细胞后,采用MTT法检测呕吐毒素对细胞增殖的抑制作用,通过荧光显微镜观察细胞形态学的变化,流式细胞术检测细胞凋亡率的变化,采用Western bloting法检测凋亡蛋白Bax、bcl-2、NF-KB、Caspase-2、3、6、8、9的表达水平。研究结果表明,呕吐毒素可显著抑制HK2细胞的增殖,具有明显的量效关系,15、30、40、60和80 μmol/L的呕吐毒素对HK2细胞的抑制率分别为61.82%、53.64%、42.43%、38.66%和36.54%;30 μmol/L的呕吐毒素处理HK2细胞0、12、24、48、72和96 h后,细胞的存活率分别为99.84%、80.51%、60.25%、52.22%、62.50%和71.34%。15、40和80 μmol/L的呕吐毒素处理细胞24 h后,细胞的凋亡率分别为2.30%、30.20%和53.50%。Western bloting结果显示呕吐毒素可上调Bax、Caspase-2、3、6、8、9蛋白表达,抑制了bcl-2和NF-KB的蛋白表达。结果表明,呕吐毒素对HK2细胞的增殖有明显的抑制作用,使得细胞发生凋亡,同时对凋亡蛋白的表达具有一定的调节作用。
[Key word]
[Abstract]
This paper investigated the effects of vomit toxin on inhibiting the proliferation and inducing the expression of apoptotic protein in HK2 cells. After different concentrations of vomit toxin acted on HK2 cells, the inhibitory effect of vomit toxin on cell proliferation was detected by MTT method, the changes of cell morphology were observed by fluorescence microscope, the changes of apoptosis rate were detected by flow cytometry, and the expression levels of apoptotic proteins Bax, Bcl-2, NF-KB, caspase-2, 3, 6, 8 and 9 were detected by Western blotting. The results showed that vomit toxin could significantly inhibit the proliferation of HK2 cells in a dose-response relationship, at the concentration of 15, 30, 40, 60 and 80 μmol/L, the inhibitory rates of vomit toxin on HK2 cells were 61.82%, 53.64%, 42.43%, 38.66% and 36.54%, respectively; The survival rates of HK2 cells treated with 30 μmol/L of vomit toxin for 0, 12, 24, 48, 72 and 96 hours were 99.84%, 80.51%, 60.25%, 52.22%, 62.50% and 71.34% respectively. The apoptosis rates of cells treated with 15, 40 and 80 μmol/L of vomit toxin for 24 hours were 2.30%, 30.20% and 53.50% respectively. The Western bloting results showed that vomit toxin upregulated Bax, Caspase-2, 3, 6, 8, 9 protein expression and inhibited the protein expression of bcl-2 and NF-KB. The results showed that vomit toxin significantly inhibited the proliferation of HK2 cells, caused apoptosis and regulated the expression of apoptotic protein.
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[基金项目]
甘肃省高等学校科研项目(2016B-024);甘肃省自然科学基金项目(18JR3RA136);国家自然科学基金地区科学基金项目(31760495)