晚期糖基化终产物对人肝癌细胞HepG2耐药性的影响-东南大学学报医学版

晚期糖基化终产物对人肝癌细胞HepG2耐药性的影响

单位：1. 中国药科大学药学院, 江苏南京 210009;
2. 东南大学附属第二医院, 江苏南京 210003;
3. 东南大学医学院, 江苏南京 210009;
4. 东南大学附属中大医院, 江苏南京 210009

分类号：R969

出版年·卷·期（页码）：2013·32·第四期（399-403）

摘要：

目的: 探讨晚期糖基化终产物(advanced glycation end products,AGEs)对肝癌细胞耐药性的影响及其机制。方法: 体外培养人肝癌细胞HepG2,以终浓度分别为100、200、400 μg·ml^-1的AGEs联合2 000 nmol·L^-1盐酸阿霉素(adramycin,ADM)处理细胞24 h,并设空白对照和ADM组进行比较。应用倒置显微镜观察不同浓度AGEs联合ADM孵育下HepG2形态学变化,流式细胞术(flow cytometry,FCM)检测细胞周期的改变,应用细胞计数试剂盒(cell counting kit-8,CCK-8)检测细胞株活性,蛋白质免疫印迹法(Western blotting)测定不同浓度AGEs作用下,HepG2细胞多药耐药基因(multidrug resistance-1,MDR1)蛋白表达情况。结果: 在2 000 nmol·L^-1ADM作用下,HepG2细胞生长停滞或死亡,而AGEs能促进细胞生长,抑制其死亡。细胞周期和细胞活性实验表明,与ADM组相比,随着AGEs浓度增加,G₁期细胞百分率显著下降,S期及G₂期细胞增加,细胞活性有上升趋势。Western blotting检测表明AGEs能增加MDR1的蛋白表达。结论: AGEs能通过促进MDR1基因的表达,降低肿瘤细胞对化疗药物的敏感性。

Objective: To investigated the effects of advanced glycation end products (AGEs) on the multidrug resistance of HepG2 cells and its mechanism. Methods: Human hepatocellular carcinoma HepG2 cells were cultured and exposed to AGEs at the concentration of 100,200 and 400 μg·ml^-1 combined with 2 000 nmol·L^-1 adramycin(ADM) for 24 hours in vitro, normal control group and ADM group were established for comparison.Inverted microscope was used to observe the change of cell morphous. The flow cytometry (FCM) was used to investigate cell cycle and cell counting kit-8 (CCK-8) assay was used to assess the cell activity. Western blotting was used to study protein expression of MDR1. Results: In cell culture medium with 2 000 nmol·L^-1 ADM, HepG2 showed arrest of growth and death, and AGEs could promote HepG2 cell growth and prevent cell death. FCM and CCK-8 experimental results showed that the percentage of cells in G₁ phase declined significantly, S phase and G₂ phase increased, cell activity was significantly higher than ADM group, the phenomena were more obvious when AGEs concentration increasing.Western blotting experimental results showed that AGEs could increase protein expression of MDR1.Conclusion: AGEs could promote MDR of tumor cells to chemotherapy by increasing MDR1 gene expression.

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