胃铋镁中药组分对胃癌细胞周期的调节和裸鼠体内肿瘤抑制的实验研究

Effects of Chinese medicine portions of Weibimei on cell cycle regulation and tumor inhibition in nude mice

目的研究胃铋镁中药组分(弗朗鼠李皮、茴香、石菖蒲、甘草和芦荟)对胃癌细胞周期的调节作用和对胃癌移植瘤的生长抑制作用。方法将胃铋镁和胃铋镁中药总组分及其组分中弗朗鼠李皮、茴香、石菖蒲、甘草、芦荟分别作用于胃癌细胞SGC-7901,MTT法检测各处理组细胞的增殖状态;流式细胞仪检测细胞周期;Western blot检测细胞周期相关蛋白的表达。构建裸鼠SGC-7901胃癌移植瘤模型,研究胃铋镁及其中药总组分及其组分对胃癌移植瘤的生长抑制作用;免疫组化检测STAT3信号通路相关蛋白STAT3、p-STAT3、Bcl-2、Cyclin D1和Survivin的表达情况。结果胃铋镁中药总组分及其组分中的弗朗鼠李皮、茴香和石菖蒲对SGC-7901细胞的抑制率显著高于对照组(P<0.05);流式检测和Western blot结果显示,胃铋镁、胃铋镁中药总组分及其组分中弗朗鼠李皮、茴香和石菖蒲均可以使胃癌细胞阻滞于G1期(P<0.05),并能够有效下调细胞周期蛋白Cyclin A、B、D、E的表达;但甘草和芦荟对胃癌细胞的增殖抑制和细胞周期阻滞作用不显著。荷瘤裸鼠实验结果显示,胃铋镁中药总组分组、弗朗鼠李皮组、茴香组和石菖蒲组对胃癌细胞的生长抑制作用显著强于对照组和三联药(三种市售胃炎胃溃疡类药物联合用药)组(P<0.05),并可以抑制STAT3的磷酸化,下调STAT3信号通路靶基因Bcl-2、Cyclin D1和Survivin的表达。结论胃铋镁及其中药组分中的弗朗鼠李皮、茴香和石菖蒲可以显著抑制胃癌细胞SGC-7901增殖,阻滞细胞周期于G1期,并可以在体内抑制荷瘤裸鼠肿瘤的生长。

Objective To study the effects of Weibimei and its Chinese medicine portions（ cortex frangulae,fennel powder,acorus tatarinowii,glycyrrhizae and asparagus） on cell cycle regulation in gastric cancer cells and growth inhibition in gastric cancer Xenografts. Methods Gastric cancer SGC-7901 cells were normally cultured and treated with Weibimei,extractum glycyrrhizae,cortex frangulae,fennel powder,acorus tatarinowii and asparagus. Then,the proliferation of SGC-7901 cells were detected by MTT assay,their effect on SGC-7901 cell cycle were measured by flow cytometry,and the expression of cell cycle related proteins were examined by Western blot. Animal models of SGC-7901 cells xenografts in nude mice were constructed to evaluate the growth inhibition effects of Weibimei and its Chinese medicine portions,and the expression of STAT3,p-STAT3,Cyclin D1,Bcl-2 and Survivin in gastric tumors were detected by immunohistochemical assays. Results Weibimei and its three kinds of Chinese medicine portions,cortex frangulae,fennel and acorus tatarinowii could obviously inhibit the proliferation of SGC-7901 cells,compared to control group,with a statistical significant difference（ P〈0. 05）. The cell cycle of SGC-7901 was arrested at G1 phase with cortex frangulae,fennel and acorus tatarinowii treatment,the expression of Cyclin A,B,D,E were decreased significantly（ P〈0. 05）. However,no significant effects was found in glycyrrhizae or asparagus group. Xenografts tumor sizes in Weibimei,cortex frangulae,fennel or acorus tatarinowi groups were much smaller than that in saline group or Triple Therapy group（ three-kinds-drugs-combination for the treatment of gastric ulcer）（ P〈0. 05）. Xenografts experiments showed that the tumor growth in nude mice in Weibimei,cortex frangulae,fennel or acorus tatarinowi groups were significantly smaller than that in control group or in Triple Therapy Group（ P〈0. 05）,and phosphorylated STAT3,and STAT3 signaling targeted genes,including Bcl-2,Cyclin D1 and Survivin were all significantly down-regulated by immunostaining. Conclusion Weibimei and its Chinese medicine portions including cortex frangulae,fennel and acorus tatarinowii can could significantly inhibit the growth of SGC-7901 cells proliferation and arrested the cell cycle of G1 phase in vitro,and inhibit gastric cancer xenografts in nude mice in vivo.