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
目的探讨环氧合酶-1(COX-1)、环氧合酶-2(COX-2)基因多态性与阿司匹林抵抗相关性。方法将96例缺血性脑卒中患者分为阿司匹林敏感组87例和阿司匹林非敏感组9例。2组患者口服100 mg阿司匹林至少一周。用光比浊法对患者的血小板聚集度进行...目的探讨环氧合酶-1(COX-1)、环氧合酶-2(COX-2)基因多态性与阿司匹林抵抗相关性。方法将96例缺血性脑卒中患者分为阿司匹林敏感组87例和阿司匹林非敏感组9例。2组患者口服100 mg阿司匹林至少一周。用光比浊法对患者的血小板聚集度进行测定,用Sequenom Mass A RRAY i PLEX基因型分析技术进行COX-1、COX-2的基因型检测,分析COX-1、COX-2基因多态性与阿司匹林的相关性。结果阿司匹林非敏感组患者的既往卒中史比例66.7%(6/9)显著高于阿司匹林敏感组21.9%(19/87)(P<0.05)。2组患者的COX-1(rs1330344,rs3842788)基因型分布差异有统计学意义(P<0.05),但COX-1(rs5788)、COX-2(rs20417)基因型分布差异无统计学意义(P>0.05)。COX-1(rs5788,rs1330344,rs3842788)、COX-2(rs20417)等位基因及基因型频率4个单核苷酸基因多态性均符合Hardy-Weinberg平衡(P>0.05)。结论 COX-1(rs3842788,rs1330344)GG型与阿司匹林抵抗密切相关。展开更多
Objective To transfect antisense vector of human cyclooxygenase-2 (COX-2) gene into COX-2 highly expressing chol-angiocarcinoma cell line QBC939 and explore its biological activities and role in carcinogenesis. Method...Objective To transfect antisense vector of human cyclooxygenase-2 (COX-2) gene into COX-2 highly expressing chol-angiocarcinoma cell line QBC939 and explore its biological activities and role in carcinogenesis. Methods QBC939 cells were transfected with antisense vector of human COX-2 gene using LipoVecTM transfecting te-chnique. Transfected cells were selected with G418; COX-2 mRNA was examined using reverse transcription polymerase chain reaction (RT-PCR) and COX-2 protein expression was detected by immunocytochemistry using isozyme selective anti-bodies. The proliferative status of transfected cells was measured by using methabenzthiazuron (MTT) assay; Cell cycle and apoptosis were analyzed by using flow cytometry. Results RT-PCR showed a lower COX-2 mRNA level in antisense vector transfected cells and immunocytochemistry showed a weaker COX-2 protein expression in antisense vector transfected cells. The antisense vector transfected cells proli-ferative index decreased significantly (P< 0.01), the percentage of S phase decreased remarkably (P< 0.05) in antisense vec-tor transfected cells (9.27% ±1.91%) compared with that in QBC939 cells without transfection(16.35% ±2.87%), and the percentage of G0/G1 phase increased remarkably (P< 0.05) in antisense vector transfected cells (75.16%±4.13%) compared with that in QBC939 cells without transfection (57.31% ±10.16%). Transfection with antisense vector of human COX-2 gene had no significant influence on the apoptosis in QBC939 cells (P> 0.05). Conclusion Transfection with antisense vector of human COX-2 gene could inhibit the proliferation of human cholan-giocarcinoma QBC939 cells.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.
文摘目的探讨环氧合酶-1(COX-1)、环氧合酶-2(COX-2)基因多态性与阿司匹林抵抗相关性。方法将96例缺血性脑卒中患者分为阿司匹林敏感组87例和阿司匹林非敏感组9例。2组患者口服100 mg阿司匹林至少一周。用光比浊法对患者的血小板聚集度进行测定,用Sequenom Mass A RRAY i PLEX基因型分析技术进行COX-1、COX-2的基因型检测,分析COX-1、COX-2基因多态性与阿司匹林的相关性。结果阿司匹林非敏感组患者的既往卒中史比例66.7%(6/9)显著高于阿司匹林敏感组21.9%(19/87)(P<0.05)。2组患者的COX-1(rs1330344,rs3842788)基因型分布差异有统计学意义(P<0.05),但COX-1(rs5788)、COX-2(rs20417)基因型分布差异无统计学意义(P>0.05)。COX-1(rs5788,rs1330344,rs3842788)、COX-2(rs20417)等位基因及基因型频率4个单核苷酸基因多态性均符合Hardy-Weinberg平衡(P>0.05)。结论 COX-1(rs3842788,rs1330344)GG型与阿司匹林抵抗密切相关。
文摘Objective To transfect antisense vector of human cyclooxygenase-2 (COX-2) gene into COX-2 highly expressing chol-angiocarcinoma cell line QBC939 and explore its biological activities and role in carcinogenesis. Methods QBC939 cells were transfected with antisense vector of human COX-2 gene using LipoVecTM transfecting te-chnique. Transfected cells were selected with G418; COX-2 mRNA was examined using reverse transcription polymerase chain reaction (RT-PCR) and COX-2 protein expression was detected by immunocytochemistry using isozyme selective anti-bodies. The proliferative status of transfected cells was measured by using methabenzthiazuron (MTT) assay; Cell cycle and apoptosis were analyzed by using flow cytometry. Results RT-PCR showed a lower COX-2 mRNA level in antisense vector transfected cells and immunocytochemistry showed a weaker COX-2 protein expression in antisense vector transfected cells. The antisense vector transfected cells proli-ferative index decreased significantly (P< 0.01), the percentage of S phase decreased remarkably (P< 0.05) in antisense vec-tor transfected cells (9.27% ±1.91%) compared with that in QBC939 cells without transfection(16.35% ±2.87%), and the percentage of G0/G1 phase increased remarkably (P< 0.05) in antisense vector transfected cells (75.16%±4.13%) compared with that in QBC939 cells without transfection (57.31% ±10.16%). Transfection with antisense vector of human COX-2 gene had no significant influence on the apoptosis in QBC939 cells (P> 0.05). Conclusion Transfection with antisense vector of human COX-2 gene could inhibit the proliferation of human cholan-giocarcinoma QBC939 cells.
