摘要
目的:观察低氧下启膈散和顺铂抑制食管癌细胞增殖的协同效应,从miR-21及其靶基因角度探讨其机制。方法:制备含药血清,噻唑蓝(MTT)比色法检测启膈散和顺铂单用或联合应用对食管癌细胞EC9706增殖的影响,实时荧光定量PCR(Real-time PCR)检测启膈散和顺铂单用或两者联合应用对食管癌细胞miR-21,程序死亡蛋白4(PDCD4),磷酸酶和张力蛋白类似物(PTEN)mRNA表达的影响,免疫印迹法(Western blot)检测PDCD4,PTEN蛋白表达。结果:启膈散在浓度为8%,10%与1 mg·L-1顺铂合用具有协同作用,其两药相互作用系数(CDI)分别为0.90,0.94。顺铂高浓度组,联合高、低浓度组miRNA-21表达量明显低于空白组(P<0.05);联合高、低浓度组miRNA-21表达量显著低于同浓度顺铂组(P<0.01),且联合高浓度组低于联合低浓度组(P<0.05)。顺铂高浓度、启膈散组PDCD4 mRNA表达量明显低于空白组(P<0.05),而联合高、低浓度组PDCD4,PTEN mRNA表达量明显高于空白组(P<0.05);联合高、低浓度组PDCD4,PTEN mRNA明显高于同浓度顺铂组(P<0.01),且联合高浓度组PTEN mRNA表达高于联合低浓度组(P<0.01)。与空白组比较,联合高、低浓度组PTEN蛋白表达明显升高(P<0.05),且联合高、低浓度组PTEN蛋白显著高于同浓度顺铂组(P<0.01);顺铂高、低浓度组,联合高、低浓度组PDCD4明显高于空白组(P<0.05),联合高浓度组明显高于同浓度顺铂组和联合低浓度组(P<0.05,P<0.01)。结论:启膈散含药血清和顺铂在抑制食管癌细胞EC9706增殖方面具有协同作用,其机制可能与通过抑制miR-21表达从而升高PDCD4和PTEN基因表达相关。
Objective： To investigate the synergistic effect of Qige San and cisplatin in inhibiting proliferation of esophageal carcinoma cells in hypoxia,in order to explore the mechanism from the perspective of miR-21 and its target genes. Method： Drug-containing serum was prepared. The single or combined effects of Qige San and cisplatin on cells activation were tested by 3-（ 4,5-dimethyl-2-thiazolyl）-2,5-diphenyl-2-H-tetrazolium bromid（ MTT） assays. Apoptosis and cell cycle arrest induced by Qige San and cisplatin were detected by flow cytometry. MiR-21,programmed cell death 4（ PDCD4）,phosphatase and tensin homolog（ PTEN） mRNA expressions were detected by Real-time PCR,while PDCD4 and PTEN protein expression were detected by Western blot. Result： Qige San in concentrations of 8% and 10% combined with 1 mg·L- 1cisplatin has a synergistic effect in inhibiting proliferation of esophageal carcinoma cells,with the coefficients of drug in interaction（ CDI） of 0. 90,0. 94,respectively. The expressions of miRNA-21 in cisplatin high concentration group,and combination high and low concentration group were significantly lower than that in the control group（ P 〈0. 05）. The expression of miRNA-21 in combination high and low concentration group was significantly lower than that of cisplatin groups with the same concentration（ P 〈0. 01）,and the low concentration group（ P 〈0. 05）. The expression of PDCD4 mRNA was significantly lower in the cisplatin high concentration group and the Qige San group than in control group（ P 〈0. 05）,whereas the expression of PTEN and PDCD4 mRNA in combination high and low concentration group was significantly higher than that in control group（ P 〈0. 05）,and PDCD4,PTEN mRNA expressions in combination high and low concentration group was significantly higher than that of cisplatin groups with the same concentration（ P 〈0. 01）,and PTEN mRNA in high concentration combination group was higher than that in lower concentration combination group（ P
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2016年第15期134-138,共5页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金面上项目(81373535)
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)
