2逆轉(zhuǎn)HepG2細(xì)胞對(duì)5-氟尿嘧啶(5-Flu)耐藥性的作用。方法 構(gòu)建對(duì)5-Flu耐藥的HepG2細(xì)胞系(HepG2-5-Flu),MTT法分別測(cè)定人參皂苷Rh2和5-Flu(二者質(zhì)量濃度均為15.63、31.25、62.50、125.00、250.00、500.00、1 000.00、2 000.00 μg/mL)、人參皂苷Rh2和5-Flu不同比例(5∶1、3∶1、1∶1、1∶3、1∶5)的混合藥物(總濃度同單給藥)對(duì)HepG2-5-Flu細(xì)胞存活率的影響,利用協(xié)同作用系數(shù)(CI)獲得人參皂苷Rh2和5-Flu的最佳協(xié)同作用濃度;通過Annexin V/PI雙染色流式細(xì)胞術(shù)檢測(cè)5-Flu(375 μg/mL)、人參皂苷Rh2+5-Flu(人參皂苷Rh2∶5-Flu=1∶3,濃度為125、375 μg/mL)對(duì)細(xì)胞凋亡影響;實(shí)時(shí)熒光定量PCR(qRT-PCR)法檢測(cè)細(xì)胞P-糖蛋白(P-gp)和多藥耐藥相關(guān)蛋白(MRP)的mRNA表達(dá)水平。結(jié)果 人參皂苷Rh2對(duì)HepG2-5-Flu細(xì)胞的半數(shù)抑制濃度(IC50)為223.3 μg/mL,而5-Flu的IC50為8.34 μg/mL。人參皂苷Rh2與5-Flu濃度比為5∶1、3∶1、1∶1、1∶3、1∶5時(shí),混合藥物對(duì)細(xì)胞的IC50分別為324.7、224.0、219.0、135.6、105.9 μg/mL。人參皂苷Rh2和5-Flu比例為5∶1時(shí),二者沒有協(xié)同作用(CI>1);當(dāng)人參皂苷Rh2和5-Flu比例為3∶1和1∶1時(shí),藥物濃度大于62.5 μg/mL時(shí)才有較小的協(xié)同作用(0.62和5-Flu比例為1∶3和1∶5時(shí),大多數(shù)藥物濃度下都有較好的協(xié)同作用(02+5-Flu組細(xì)胞壞死、早期凋亡、晚期凋亡的比例均顯著升高,活細(xì)胞比例顯著下降(P<0.05、0.01、0.001),MDR1和P-gp 2個(gè)耐藥相關(guān)蛋白的mRNA表達(dá)顯著下調(diào)(P<0.01)。結(jié)論 人參皂苷Rh2能夠逆轉(zhuǎn)HepG2細(xì)胞對(duì)5-Flu的耐藥性,機(jī)制可能與下調(diào)MDR1和P-gp表達(dá)相關(guān)。;Objective To study anti-liver cancer efficacy and overcome the drug resistance of 5-fluorouracil (5-Flu) that synergistically enhanced by ginsenoside Rh2. Methods HepG2 cell line (HepG2-5-Flu) resistant to 5-Flu was constructed, Ginsenoside Rh2 and 5-Flu (15.63, 31.25, 62.50, 125.00, 250.00, 500.00, 1 000.00, 2 000.00 μg/mL) were determined by MTT method, different proportions (5:1, 3:1, 1:1, 1:3, 1:5) of ginsenoside Rh2 and 5-Flu (the total concentration was the same as single administration) on the survival rate of HepG2-5-Flu cells, and the optimal synergistic concentration of Ginsenoside Rh2 and 5-FLu was obtained by using the synergistic coefficient. The effects of 5-Flu (375 μg/mL) and ginsenoside Rh2+ 5-Flu (Ginsenoside Rh2:5-Flu=1:3, 125, 375 μg/mL) on apoptosis were detected by Annexin V/PI double staining flow cytometry. The mRNA expression levels of P-gp and MRP were detected by real-time quantitative PCR (qRT-PCR). Results The 50% inhibitory concentration (IC50) of Ginsenoside Rh2 against HepG2-5-Flu cells was 223.3 μg/mL, and that of 5-Flu was 8.34 μg/mL. The IC50 of ginsenoside Rh2 and 5-Flu were 324.7, 224.0, 219.0, 135.6 and 105.9 μg/mL when the concentration ratio of ginsenoside Rh2 and 5-flu was 5:1, 3:1, 1:1, 1:3 and 1:5, respectively. When the ratio of ginsenosides Rh2 and 5-Flu was 5:1, there was no synergistic effect (CI > 1). When the ratio of ginsenoside Rh2 and 5-Flu was 3:1 and 1:1, the concentration of ginsenoside Rh2and 5-Flu was greater than 62.5 μg/mL (0.6 < CI < 1). When the ratio of ginsenoside Rh2 and 5-Flu was 1:3 and 1:5, most drugs had a good synergistic effect (0 2 + 5-Flu group were significantly increased, and the percentages of living cells were significantly decreased (P < 0.05, 0.01, 0.001), and the mRNA expressions of MDR1 and P-GP were significantly down-regulated (P < 0.01). Conclusion Ginsenoside Rh2 can reverse the resistance of HepG2 cells to 5-Flu, and the mechanism may be related to the down-regulation of MDR1 and P-GP expression."/> 2;5-fluorouracil;liver cancer;synergistical effect;drug resistance"/>
Objective To study anti-liver cancer efficacy and overcome the drug resistance of 5-fluorouracil (5-Flu) that synergistically enhanced by ginsenoside Rh2. Methods HepG2 cell line (HepG2-5-Flu) resistant to 5-Flu was constructed, Ginsenoside Rh2 and 5-Flu (15.63, 31.25, 62.50, 125.00, 250.00, 500.00, 1 000.00, 2 000.00 μg/mL) were determined by MTT method, different proportions (5:1, 3:1, 1:1, 1:3, 1:5) of ginsenoside Rh2 and 5-Flu (the total concentration was the same as single administration) on the survival rate of HepG2-5-Flu cells, and the optimal synergistic concentration of Ginsenoside Rh2 and 5-FLu was obtained by using the synergistic coefficient. The effects of 5-Flu (375 μg/mL) and ginsenoside Rh2+ 5-Flu (Ginsenoside Rh2:5-Flu=1:3, 125, 375 μg/mL) on apoptosis were detected by Annexin V/PI double staining flow cytometry. The mRNA expression levels of P-gp and MRP were detected by real-time quantitative PCR (qRT-PCR). Results The 50% inhibitory concentration (IC50) of Ginsenoside Rh2 against HepG2-5-Flu cells was 223.3 μg/mL, and that of 5-Flu was 8.34 μg/mL. The IC50 of ginsenoside Rh2 and 5-Flu were 324.7, 224.0, 219.0, 135.6 and 105.9 μg/mL when the concentration ratio of ginsenoside Rh2 and 5-flu was 5:1, 3:1, 1:1, 1:3 and 1:5, respectively. When the ratio of ginsenosides Rh2 and 5-Flu was 5:1, there was no synergistic effect (CI > 1). When the ratio of ginsenoside Rh2 and 5-Flu was 3:1 and 1:1, the concentration of ginsenoside Rh2and 5-Flu was greater than 62.5 μg/mL (0.6 < CI < 1). When the ratio of ginsenoside Rh2 and 5-Flu was 1:3 and 1:5, most drugs had a good synergistic effect (0 2 + 5-Flu group were significantly increased, and the percentages of living cells were significantly decreased (P < 0.05, 0.01, 0.001), and the mRNA expressions of MDR1 and P-GP were significantly down-regulated (P < 0.01). Conclusion Ginsenoside Rh2 can reverse the resistance of HepG2 cells to 5-Flu, and the mechanism may be related to the down-regulation of MDR1 and P-GP expression.