摘要
目的:观察启膈散含药血清联合顺铂对人食管癌细胞EC9706的增殖效应,从miR-21探讨其作用机制。方法:制备启膈散含药血清,MTT法筛选其最佳作用浓度;PCR检测miR-21、PDCD4 mRNA和PTEN mRNA的表达情况;Western-blot检测蛋白PDCD4、PTEN的表达水平。结果:5%含药血清的浓度作用效果最为显著(q>1)。与5%对照血清组比较,除5%含药血清PDCD、PTEN mRNA和0.5μg/m L顺铂组的PTEN mRNA表达量下降外,其余各组的miR-21、PDCD4 mRNA和PTEN mRNA表达均升高(P<0.05);顺铂联合含药血清后,两组miR-21表达和联合高浓度顺铂组的PDCD4 mRNA均比其单用下降(P<0.05),而两组PTEN mRNA和联合低浓度组的PDCD4 mRNA的表达则升高(P<0.05)。与对照组相比,各组PDCD4蛋白表达量升高P<0.05;高浓度顺铂联合含药血清组比单用高顺铂组PDCD4蛋白表达量升高(P<0.05),低浓度顺铂的两组变化不明显(P>0.05);两个浓度顺铂联合含药血清时PTEN的表达量均高于单独使用顺铂(P<0.05)。结论:启膈散能够增加EC9706对顺铂的敏感性,可能是通过miR-21与其调控的下游基因PDCD4与PTEN共同参与完成。
Objective ： To oberserve the proliferation of human esophageal cancer cells EC9706 effected by Qige Powder -containing serum combined cisplatin and explore its mechanism by the miR -21. Methods： Qige Powder -containing serum was prepared and MTT method was utilized to screen out the most effective drug - containing serum concentrations. PCR was utitlized to detect the expressions of miR - 21, PDCD4 mRNA and PTEN mRNA. Western blot was used to test the protein levels of PDCD4 and PTEN. Results： The synergistic effect of 5% medicated serum with cisplatin was the most significant（q 〉 1 ）. Compared with 5% control serum group, in addition to PDCD, PTEN mRNA in the 5% medica- ted serum group and the PTEN mRNA expression in 0.5 μg/mL cisplatin group decreased. The miR - 21, PDCD4 mR- NA and PTEN mRNA expressions in the rest of the groups were higher（P 〈 0. 05 ）. The miR - 2t expression in two cispl- atin combined with medicated serum groups（CCS） and were lower than those of groups with the drugs alone（ P 〈 0.05 ） while PTEN mRNA in two CCS groups and PDCD4 mRNA in low concentration CCS group were increased（ P 〈 0. 05 ）. Compared with the control group, PDCD4 protein expression of each group was rised（P 〈 0. 05 ）. PDCD4 protein expres- sion from high concentrations CCS group was higher than that of the high concentration cisplatin alone （ P 〈 0.05 ）. The expression of PTEN in two concentrations CCS were higher than that of cisplatin alone （ P 〈 0. 05 ）. Conclusion ： Qige Powder can heighten the sensitivity of EC9706 to cisplatin, which may be completed by miR -21 and its target genes PD- CD4 and PTEN together.
出处
《中华中医药学刊》
CAS
北大核心
2017年第12期3098-3101,共4页
Chinese Archives of Traditional Chinese Medicine
基金
国家自然科学基金面上项目(81373535)
河南省高等学校重点科研项目(15A310020)
河南省科技攻关计划项目(142102310038)
河南省教育厅科学技术研究重点项目(14A360025)
河南中医药大学省属科研专项项目
作者简介
吴耀松（1976-），女，湖南娄底人，副教授，硕士，研究方向：肿瘤与防治。;通讯作者：陈玉龙（1973-），男，河南平舆人，教授，博士，研究方向：肿瘤病机与防治。E-mail：cyl72621@163．com。
