Objective： To evaluate the underlying mechanism of Jianpi Jiedu Recipe （健脾解毒方, JJR） in the reversion of multidrug resistance concerning colorectal cancer in vitro and in vivo. Methods： Mice were treated orall...Objective： To evaluate the underlying mechanism of Jianpi Jiedu Recipe （健脾解毒方, JJR） in the reversion of multidrug resistance concerning colorectal cancer in vitro and in vivo. Methods： Mice were treated orally with JJR at a daily 4.25 g/（kg.day） or injected with vinblastine （VCR） 2.5 mg/（kg day） for 3 weeks after having been inoculated with HCT8N cells; tumor tissues were assayed by hematoxylin and eosin staining. Firstly, the effects of JJR on the expression of cyclooxygenase-2 （COX-2） were tested by real-time polymerase chain reaction （PCR） technique and COX-2 gene silenced by siRNA. Secondly, the variation of intracellular concentration of oxaliplatin （L-OHP） was evaluated by the inductively coupled plasma mass spectroscopy （ICP- MS） in HCT8N and its COX-2 siRNA cells; the concentration of J JR combined with chemotherapeutic drugs and the reverse effect of multidrug resistance （MDR） in HCT8N cells was evaluated by the MTT assay. Thirdly, real-time quantitative PCR and Western blot analysis were used to detect the multidrug resistance gene 1 （MDR1） mRNA and P-gp expression. Results： JJR had an inhibitory effect on the growth of tumors in vivo, and it, in combination with chemotherapeutic drugs, could reverse the drug-resistance of HCT8N cells and increase the sensitivity of HCT8N cells to VCR, DDP, 5-Fu, and THP. ICP-MS results showed that JJR could increase the concentration of drugs in HCT8/V cells （P〈0.01）. Furthermore, it was shown that JJR could reverse drug resistance of colorectal cancer cells by decreasing MDR1 expression and P-gp level via downregulation of COX-2, which has been represented as one of the major mechanisms that contributes to the MDR phenotype （P〈0.01）. Conclusion： JJR reversed multidrug resistance and enhanced the sensitivity to chemotherapy, which could be attributed to the down-regulation of COX-2 in MDRl/P-gp-mediated MDR colorectal cancer after chemotherapy.展开更多
基金Supported by the National Natural Science Foundation of China(No.81202812)Science and Technology Commission of Shanghai Municipality(No.10ZR1427400)+1 种基金Program of Shanghai MunicipalEducation Commission(No.09YZ132,2011JW57)Shanghai Municipal Health Bureau(No.2011ZJ030,20114Y013)
文摘Objective： To evaluate the underlying mechanism of Jianpi Jiedu Recipe （健脾解毒方, JJR） in the reversion of multidrug resistance concerning colorectal cancer in vitro and in vivo. Methods： Mice were treated orally with JJR at a daily 4.25 g/（kg.day） or injected with vinblastine （VCR） 2.5 mg/（kg day） for 3 weeks after having been inoculated with HCT8N cells; tumor tissues were assayed by hematoxylin and eosin staining. Firstly, the effects of JJR on the expression of cyclooxygenase-2 （COX-2） were tested by real-time polymerase chain reaction （PCR） technique and COX-2 gene silenced by siRNA. Secondly, the variation of intracellular concentration of oxaliplatin （L-OHP） was evaluated by the inductively coupled plasma mass spectroscopy （ICP- MS） in HCT8N and its COX-2 siRNA cells; the concentration of J JR combined with chemotherapeutic drugs and the reverse effect of multidrug resistance （MDR） in HCT8N cells was evaluated by the MTT assay. Thirdly, real-time quantitative PCR and Western blot analysis were used to detect the multidrug resistance gene 1 （MDR1） mRNA and P-gp expression. Results： JJR had an inhibitory effect on the growth of tumors in vivo, and it, in combination with chemotherapeutic drugs, could reverse the drug-resistance of HCT8N cells and increase the sensitivity of HCT8N cells to VCR, DDP, 5-Fu, and THP. ICP-MS results showed that JJR could increase the concentration of drugs in HCT8/V cells （P〈0.01）. Furthermore, it was shown that JJR could reverse drug resistance of colorectal cancer cells by decreasing MDR1 expression and P-gp level via downregulation of COX-2, which has been represented as one of the major mechanisms that contributes to the MDR phenotype （P〈0.01）. Conclusion： JJR reversed multidrug resistance and enhanced the sensitivity to chemotherapy, which could be attributed to the down-regulation of COX-2 in MDRl/P-gp-mediated MDR colorectal cancer after chemotherapy.
