千金子炮制前后对人肾癌786-0细胞增殖及特异抗原G250基因表达的影响

Effects of caper euphorbia seed before and after processing on the proliferation of human renal carcinoma 786-0 cells and the gene expression of special antigen G250

目的:比较千金子炮制前后对人肾癌786-0细胞增殖及相关抗原G250基因表达的影响。方法:采用噻唑蓝(MTT)比色法测定千金子炮制前后及不同部位对人肾癌786-0细胞体外生长抑制情况;采用实时荧光定量PCR技术检测千金子炮制前后对肾癌相关抗原G250表达的影响。结果:千金子霜、种仁、种子、种皮甲醇提取物作用96h可显著抑制786-0细胞的增殖,且除千金子种皮外呈浓度依赖性。实时荧光定量PCR实验结果显示,与空白对照组比较,千金子种子高、中、低3个浓度组G250基因的表达显著升高,而千金子霜组和千金子种仁组G250基因的表达则降低。结论:千金子有较强的抑制786-0细胞增殖作用,且炮制制霜后抑制作用显著提高,千金子种子抑制癌细胞增殖作用机制是上调肾癌特异抗原G250的表达。

Objective：To observe the effects of caper euphorbia seed before and after processing on the proliferation of human renal carcinoma 786-0 cells and the gene expression of special antigen G250.Methods：The inhibitory effect of caper euphorbia seed before and after processing on the proliferation of human renal carcinoma 786-0 cells was detected by MMT.The effect of caper euphorbia seed before and after processing on the gene expression of special antigen G250 was detected by real-time fluorescent quantitative PCR（real-time PCR）.Results：The methanolic extracts from semen eupoarbiae pulveratum,kernel,seeds,and seed-coat could all significantly inhibit the proliferation of 786-0 cells after 96 hours and the action showed concentration dependence except for the extract from seed-coat.Real-time PCR demonstrated that compared with the control group,the expression of G250 gene in the caper euphorbia seed high dose group,caper euphorbia seed middle dose group and caper euphorbia seed low dose group was all significantly increased;However,the expression of G250 gene in the semen eupoarbiae pulveratum group and kernel group was decreased,compared with the control group.Conclusions：Caper euphorbia seed have a strong inhibitory effect on the proliferation of 786-0 cells,which could improve dramatically after being made into frost form.Its mechanism might be related to up-regulate the expression of specific antigen G250.However,the underlying mechanism that caper euphorbia seed could efficiently inhibit renal cell carcinoma would require further study.