富硒蛹虫草对非小细胞肺癌细胞系NCI-H1299和NCI-H1975增殖凋亡的影响研究

Study on the effect of selenium enriched cordyceps militaris on proliferation and apoptosis of non-small cell lung cancer cell lines NCI-H1299 and NCI-H1975

目的观察富硒蛹虫草对非小细胞肺癌(NSCLC)细胞系NCI-H1299和NCI-H1975增殖和凋亡的影响及机制。方法将不同浓度的富硒蛹虫草作用于体外培养的NCI-H1299和NCI-H1975细胞,采用CCK-8实验检测富硒蛹虫草对NCI-H1299和NCI-H1975细胞增殖抑制率的影响;流式细胞术检测细胞凋亡率;实时荧光定量聚合酶链式反应(Real-time qPCR)和Western Blot分别检测促凋亡因子Bax、抗凋亡蛋白Bcl-2、TP53基因mRNA和蛋白,PARP的mRNA和cleaved PARP蛋白表达情况。结果富硒蛹虫草水提物对NCI-H1299和NCI-H1975增殖有明显的抑制作用,且呈浓度依赖性;富硒蛹虫草以浓度依赖性促进NCI-H1299和NCI-H1975的凋亡;促凋亡因子Bax的mRNA和蛋白表达上调,而抗凋亡蛋白Bcl-2的mRNA和蛋白表达下调;TP53仅在富硒蛹虫草浓度最高时表现出蛋白表达量下降;PARP的mRNA表达水平在富硒蛹虫草浓度最高时变化不明显,但cleaved PARP的蛋白表达随着富硒蛹虫草的浓度升高而增加。结论富硒蛹虫草能显著抑制NCI-H1299和NCI-H1975增殖,促进细胞凋亡,其机制可能通过调节Bcl-2家族蛋白,上调cleaved PARP的蛋白表达水平实现。

Objective To observe the effect and mechanism of selenium enriched cordyceps militaris on proliferation and apoptosis of non-small cell lung cancer(NSCLC) cell lines NCI-H1299 and NCI-H1975. Methods Different concentrations of selenium enriched cordyceps militaris were applied to NCI-H1299 and NCI-H1975 cells in vitro. CCK-8 experiment was used to detect the effect of selenium enriched cordyceps militaris on the proliferation inhibition rate and half inhibition concentration(IC50) of NCI-H1299 and NCI-H1975 cells. Flow cytometry was used to detect the apoptosis rate. Real-time quantitative polymerase chain reaction(Real-time qPCR) and Western Blot were used to detect the expression of apoptogenic factor Bax, apoptogenic factor Bcl-2, TP53 gene mRNA and protein, PARP mRNA and cleaved PARP protein. Results The aqueous extract of selenium enriched cordyceps militaris inhibited the proliferation of NCI-H1299 and NCI-H1975 in a concentration dependent manner. The selenium enriched cordyceps militaris can promote the apoptosis of NCI-H1299 and NCI-H1975 in a concentration dependent manner. The mRNA and protein expression of apoptogenic factor Bax was up-regulated, while that of apoptogenic factor Bcl-2 was down-regulated. The TP53 protein expression decreased only at the highest concentration of selenium enriched cordyceps militaris. The PARP mRNA expression did not change significantly at the highest concentration of selenium enriched cordyceps militaris, but protein expression of cleaved PARP increased with the increase of selenium enriched cordyceps militaris. Conclusion The selenium enriched cordyceps militaris can significantly inhibit the proliferation of NCI-H1299 and NCI-H1975, and promote cell apoptosis. The mechanism may be realized by regulating Bcl-2 family protein and up-regulating the protein expression level of cleaved PARP.