槲寄生碱影响Ras蛋白表达抑制肺癌H358细胞增殖

Inhibition effects of mistletoe alkali on the expression of K-Ras and the proliferation of H358 cells

目的研究槲寄生碱对非小细胞肺癌H358细胞的抑制作用及其相关作用机制。方法采用MTT法检测槲寄生碱对H358细胞增殖的抑制作用;采用流式细胞仪检测槲寄生碱对H358细胞凋亡的影响;采用Western blot方法检测槲寄生碱对K-Ras、Bcl-2和Bax等蛋白表达的影响;测定caspase 3、caspase 8和caspase 9的活性;检测槲寄生碱在体内的抑瘤活性。结果槲寄生碱以质量浓度依赖的方式抑制H358细胞增殖,并能有效地诱导H358细胞凋亡;caspase 3、caspase 8和caspase 9受到了不同程度的激活;同时,槲寄生碱抑制K-Ras和Bcl-2的表达,而对Bax没有明显抑制作用;槲寄生碱在体内也表现出明显的抑瘤活性。结论槲寄生碱在体外能够抑制H358细胞增殖并诱导H358细胞凋亡,为K-Ras突变型非小细胞肺癌等肿瘤疾病的治疗奠定了实验基础。

Objective To observe the inhibitory effect of mistletoe alkali on H358 cells and to investigate the relevant molecular mechanism. Methods The inhibitory effect of mistletoe alkali on H358 cells was deter- mined by MTT method. The apoptosis of H358 cells was determined by flow cytometry. The activity of caspase 3, caspase 8 and caspase 9 was determined by using caspase activity assay kit. The protein expression level of K-Ras, Bcl-2 and Bax was determined by Western blot method. Results It was shown that mistletoe alkali could inhibit the proliferation of H358 cells in dose-dependent manner both in vitro and in vivo. It could induce the apoptosis of H358 cells. The activity of caspase 3,8,9 in H358 cells was increased while the expression of K-Ras and Bcl-2 was decreased with the increase of mistletoe alkali concentration. But the expression of Bax had no change. Conclusions Mistletoe alkali can inhibit the proliferation of H358 cells in vitro and in vivo.