加热对肝癌HepG2细胞肺耐药蛋白表达及耐药性的影响  

Effect of hyperthermia on expression of lung resistance protein and sensitivity to chemotherapeutics in HepG2 cell

作　　者：张金良[1] 陈孝平[1] 张万广[1] 任利[1] 杨盛力[1] 刘利平[1] 梁慧芳[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院肝脏外科中心,湖北武汉430030

出　　处：《中国现代普通外科进展》2009年第4期279-282,共4页Chinese Journal of Current Advances in General Surgery

基　　金：国家自然科学基金资助项目(30430670)

摘　　要：目的:研究多次加热对肝癌HepG2细胞肺耐药蛋白(lung resistance protein,LRP)表达的影响以及加热后HepG2细胞对阿霉素敏感性的变化。方法:肝癌HepG2细胞在42℃恒温水浴箱中加热,60min/次,1次/d,共10d,获取稳定生长的加热HepG2细胞。以未加热HepG2细胞为对照,应用荧光定量PCR和Western Blot检测两种细胞LRP的mRNA和蛋白水平;通过体外细胞毒实验(MTT法)观察阿霉素对两种细胞生长抑制的影响,分析加热后HepG2细胞对阿霉素敏感性的变化;应用流式细胞技术检测两种细胞内阿霉素的平均荧光强度,分析加热对细胞内药物浓度的影响。结果:加热HepG2细胞的LRP mRNA和蛋白水平较未加热HepG2细胞分别增加(4.01±0.23)和(4.67±0.36)倍,差异有统计学意义(P<0.05);未加热HepG2细胞半数抑制浓度/加热HepG2细胞半数抑制浓度=3.18;应用阿霉素1h、2h后,加热HepG2细胞内平均荧光强度分别比未加热HepG2细胞高46.1%和33.9%。结论:加热虽然使LRP表达上调,但不足以引起耐药性增加;加热能够显著提高肝癌HepG2细胞对阿霉素的敏感性,其机制可能与细胞膜通透性增加,细胞内药物浓度增高有关。Objective： To study the changes in the expresson of lung resistance protein （ERP） mRNA after multiple hyperthermia and the senstivity to Adriamycin in HepG2 cell. Methods： HepG2 cell were treated by ten repeated cycles of exposure at 42 degree for 60 minute once a day, and stable HepG2 cell was obtained after hyperthermia. Compared with control HepG2 cell without hyperthermia, Real-time PCR and Westerrt Blot Were facilitated to observe the expression of LRP gene, Growth inhibition of Adriamycin to two groups was studied by MTT assay. The sensitivity of cell to Adriamycin was analyzed. Mean fluorescence intensity of ADM were detected by flow cytometry, and the changes of hyperthermia on intracellular concentration of ADM were analyzed. Results： Compared with control HepG2 cell without hyperthermia, the mRNA and protein levels of LRP in HepG2 cell increased 4.01±0.23 and 4.67±0.36 times respectively. This difference was of statistical significance（P〈0.05） in comparison with control cells. IC50（HepG2 cell）/IC50（HepG2 cell after hyperthermia0 was 3.18 times. The mean flourescence intensity of ADM in cells after hyperthermia elvated 46.1% and 33.9% respectively in 1 hour and 2 hours in comparison with control cells. Conclusion： These results demonstrated that the expression of LRP gene was increased after hyperthermia, it was insufficient to cause an increase in drug resistance in HepG2 cell lines.

关 键 词：高温 诱发 抗药性 肿瘤 肝肿瘤 实验性 

分 类 号：R735.7[医药卫生—肿瘤] R734.2[医药卫生—临床医学]