养阴化瘀解毒方促进结肠癌HCT116细胞凋亡的实验研究

Experimental Study of Yangyin Huayu Jiedu Decoction on Promoting Colon Cancer HCT116 Cell Apoptosis

为了探讨养阴化瘀解毒方(Yangyin Huayu Jiedu Decoction,YYHYJDD)含药血清对结肠癌细胞凋亡的影响,取对数生长期的HCT116细胞并将其分为正常组、空白血清对照组以及YYHYJDD含药血清低、中、高剂量组(体积分数为5%、10%、20%),细胞分别经24 h、48 h、72 h和96 h处理后用MTS试剂盒检测其增殖情况;以处理48 h的含药血清干预细胞为研究对象,采用AOEB染色法观察细胞的凋亡情况,采用Annexin V-FITC/PI双染检测细胞的凋亡率,同时通过Western-blot检测B细胞淋巴瘤2(B cell lymphoma 2,Bcl-2)、B细胞淋巴瘤2相关X蛋白(Bcl-2-associated X protein,Bax)的表达。结果显示:与空白血清对照组相比,YYHYJDD含药血清低、中、高剂量组从48 h开始抑制细胞活性,细胞出现凋亡现象,细胞凋亡率明显升高;高剂量组下调Bcl-2蛋白的表达,中、高剂量组上调Bax蛋白的表达,中、高剂量组Bcl-2/Bax的比值明显下调。实验结果初步表明YYHYJDD含药血清可促进结肠癌HCT116细胞凋亡,其机制可能与其影响Bcl-2和Bax的表达相关。

In order to investigate the effect of Yangyin Huayu Jiedu Decoction(YYHYJDD)-containing serum on the apoptosis of colon cancer cells,HCT116 cells in the logarithmic growth phase were divided into normal group,blank serum control group and serum groups containing low,medium and high doses of YYHYJDD(at volume fractions of 5%,10%and 20%).After the cells were treated for 24 h,48 h,72 h and 96 h,their proliferation was detected by MTS kit.Subsequently,the cells treated with the drug-containing serum for 48 h were taken as the study subject,and cell apoptosis was observed by AOEB staining method,Annexin V-FITC/PI double staining was used to detect cell apoptosis rate,and Western-blot was used to detect B cell lymphoma 2(Bcl-2)and Bcl-2-associated X protein(Bax)expressions.The results showed that,compared with the blank serum control group,the serum groups containing low,medium and high doses of YYHYJDD inhibited cell activity after treatment with 48 h,with cell apoptosis appearing and the apoptosis rates significantly increased.The expression of Bcl-2 protein was down-regulated in the high-dose group,and the expression of Bax protein was up-regulated in both the middle-and high-dose groups.The Bcl-2/Bax ratio of the middle-and high-dose groups was significantly reduced.The experimental results preliminarily indicate that YYHYJDD-containing serum can promote the apoptosis of colon cancer HCT116 cells,and the mechanism may be related to its influence on the expression of Bcl-2 and Bax proteins.