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肿瘤代谢与营养电子杂志
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水飞蓟素对缺氧人肝癌HepG⁃2细胞HIF⁃1α及MDR1 表达的影响
目的 探讨水飞蓟素(SM)对缺氧条件下人肝癌HepG‐2细胞中缺氧诱导因子‐1α (HIF‐1α)和多耐药基因1(MDR1)表达 的影响。 法 不同浓度的SM(0、10、20、40 mg/L)与浓度梯度的化疗药物(多柔比星、索拉菲尼、顺铂)处理HepG‐2细胞后应用四 甲基偶氮唑蓝比色法(MTT)检测不同浓度SM对HepG‐2细胞化疗药物敏感性的影响;在缺氧条件下,分别用0、10、20、40 mg/L 的 SM处理HepG‐2细胞8 h后,应用RT-PCR检测不同浓度SM对HepG‐2细胞的HIF‐1α及MDR1 mRNA表达水平的影响,利用蛋白 质印迹法检测不同浓度SM对HepG‐2细胞的HIF‐1α及P‐糖蛋白(P‐Gp)蛋白表达水平的影响。结果 随着SM浓度的增加,HepG‐2 细胞对化疗药物多柔比星、索拉菲尼和顺铂的敏感性逐渐增强;与对照组相比,10、20、40 mg/L SM处理的HIF‐1α mRNA表达量 差异无统计学意义(P>0.05),而MDR1 mRNA表达量呈浓度依赖性降低(P<0.05),10、20、40 mg/L SM处理的HIF‐1α与P‐Gp蛋白 表达水平呈浓度依赖性降低(P<0.05)。结论 SM可能通过在转录后水平抑制HepG‐2细胞HIF‐1α的蛋白表达而降低MDR1的mRNA 与蛋白表达,从而降低肝癌细胞的耐药性。
1南充市中心医院肝胆外科,四川 南充 637000;2南充市中心医院呼吸内科,四川 南充 637000
Effects of silymarin on HIF‑1α and MDR1 expression in HepG‑2 cells under hypoxia
Objective To investigate the effect of silymarin (SM) on the expression of hypoxia-inducible factor-1 alpha (HIF?1α) and multidrug resistance 1 (MDR1) in HepG?2 cells under hypoxia. Methods After treatment of HepG?2 cells with different concentrations of SM (0, 10, 20, 40 mg/L) and concentration gradient chemotherapeutic drugs (doxorubicin, sorafenib, cisplatin), MTT assay was used to detect the effect of different concentrations of SM on the sensitivity of HepG?2 cells to chemotherapeutic drugs. Under hypoxic conditions, HepG?2 cells were treated with SM at 0, 10, 20, and 40 mg/L for 8 h, RTPCR was used to detect the effects of different concentrations of SM on the expression levels of HIF?1α and MDR1 mRNA, and Western Blot was used to detect the effects of different concentrations of SM on the protein expression levels of HIF?1α and Pglycoprotein (P?Gp) in HepG?2 cells. Results With the increase of SM concentration, the sensitivity of HepG?2 cells to the chemotherapy drugs doxorubicin, sorafenib and cisplatin gradually increased. Compared with the control group, there was no significant difference in HIF?1α mRNA expression in the 10, 20, and 40 mg/L SM treatment groups (P>0.05), while the MDR1 mRNA expression decreased in a concentration-dependent manner (P<0.05). Additionally, the HIF?1α and P?Gp protein expression levels of the 10, 20, and 40 mg/L SM treatment groups decreased in a concentration-dependent manner compared with the control group (P<0.05). Conclusion SM might reduce the expression of MDR1 by inhibiting the expression of HIF?1α in HepG?2 cells at the post-transcriptional level, thereby reducing the drug resistance of hepatocellular carcinoma cells.
Department of Hepatobiliary Surgery, Central Hospital of Nanchong, Nanchong 637000, Sichuan, China;2. Department of respiratory, Central Hospital of Nanchong, Nanchong 637000, Sichuan, China