中药QHF复方对人乳腺癌MCF-7细胞的体外抗癌效果研究

Research on In- vitro Anticancer Effect of Chinese Herbal Compound QHF on Human Breast Cancer Cell Line MCF-7

目的研究中药QHF复方对人乳腺癌MCF-7细胞的体外抗癌效果。方法人乳腺癌MCF-7细胞,华蟾素∶人参皂苷Rg3∶三七总皂苷∶香菇多糖=57∶1∶0.4∶7的比例配制中药QHF复方,配成5个浓度:QHF1、QHF2、QHF3、QHF4和QHF5。四甲基偶氮唑盐微量酶反应比色法(MTT法)检测细胞抑制率、流式细胞仪检测细胞周期、酶联免疫吸附剂测定(ELISA)法检测细胞上清液中血管内皮生长因子(VEGF)含量、Transwell实验定量观察细胞迁移和侵袭能力。结果 MTT实验结果显示:不同浓度的QHF作用于MCF-7细胞24、48、72 h后,随着QHF浓度的增加,抑制率不断增加(P<0.01);且随着时间延长,药物对细胞的抑制率逐渐增加(P<0.01)。24、48、72 h的中药QHF复方IC50值对应华蟾素浓度分别为214.3、44.8、2.8μl。作用48 h后,与正常对照组比较,QHF复方组、盐酸多柔比星组、联合用药组细胞周期位于G0/G1期和S期的比例增多,位于G2/M期的比例减少(P<0.05);与联合用药组比较,QHF复方组、盐酸多柔比星组细胞周期位于G0/G1期的比例减少,位于S期和G2/M期的比例增多(P<0.05)。ELISA结果显示:作用48 h后,与正常对照组比较,QHF1组、QHF2组、QHF3组、QHF4组和QHF5组上清液中VEGF含量均下降(P<0.05)。Transwell实验结果显示:与正常对照组(127.3±5.7)比较,QHF3组(17.7±5.0)、QHF4组(51.3±2.9)和QHF5组(63.0±6.6)穿透滤膜的MCF-7细胞数目减少(P<0.01)。结论中药QHF复方在体外可抑制人乳腺癌MCF-7细胞的增殖和迁移、侵袭能力,并能下调VEGF的表达。

Objective To investigate the in- vitro anticancer effect of Chinese herbal compound QHF on human breast cancer cell line MCF- 7.Methods Chinese toad bufotoxin,ginsenoside Rg3,panax notoginseng saponins and lentinan were mixed into each other according to the proportion of 57∶ 1∶ 0.4∶ 7 to prepare into five different concentrations of QHF compound prescriptions：QHF1,QHF2,QHF3,QHF4 and QHF5.The inhibitory effect on MCF- 7 cells were observed by MTT assay,the cell cycle was detected by flow cytometry with PI staining,the protein level of VEGF was assessed by ELISA method,the cells＇ migration and invasion were observed quantitatively by transwell assay.Results MTT assay results indicated that：after 24 h,48 h and 72 h,as the concentration of QHF in MCF- 7 cells increased,the inhibition rate also increased（P〈 0.01）,and as time increased,the drug＇s inhibition rate increased as well（P 0.01）.The IC50values of medicine QHF Compound at 24 h,48 h and 72 h corresponded to cinobufotalin concentrations of 214.3 μl,44.8 μl and 2.8 μl respectively.Compared with the control group,the cell cycle were more in G0/ G1and S phases in QHF compound group,doxorubicin hydrochloride group and the combination therapy group after 48 h,and less in cell ratio of G2/ M phase in the three groups（P〈0.05）;Compared with the combination therapy group,the QHF compound group and the doxorubicin hydrochloride group were less in G0/ G1phase,more in S and G2/ M phases,showing statistically significant differences（P〈0.05）;ELISA results showed that：after 48 h,compared with the control group,the VEGF levels in supernatants of different concentrations of Chinese herbal compound QHF groups were all decreased（P〈0.05）.Transwell results showed that：compared with the control group（127.3 ± 5.7）,the number of MCF- 7 cells which penetrated through the filter membrane significantly decreased in QHF3 group（17.7 ± 5.0）,QHF4 group（51.3 ± 2.9） and QHF5 group（63.0 ± 6.6）（F = 109.667,P〈 0.01）.Conclusion Chinese herbal compound QHF can inhibit the proliferation,migration and invasion of MCF- 7 cells,and it could also down regulate the secretion of VEGF in vitro.