论文部分内容阅读
目的:通过人工诱导建立肿瘤坏死因子相关凋亡诱导配体(TRAIL)获得性耐药的肺癌细胞系,氯化锂联合rhTRAIL作用细胞,以期了解氯化锂增敏TRAIL的现象及机制。方法:通过低剂量rhTRAIL诱导人肺大细胞癌细胞系H460,建立TRAIL耐药细胞株H460R并鉴定,应用MTT和流式细胞术分析氯化锂和rhTRAIL联合给药后亲本株与耐药株细胞增殖和凋亡差异,应用RT-PCR和Western blot检测死亡受体表达。结果:rhTRAIL处理亲本株H460和耐药株H460R后细胞存活率存在显著差异(P<0.01),IC50分别为59.2ng/ml和294.8ng/ml。60ng/ml rhTRAIL处理耐药株及亲本株后平均细胞凋亡比例存在显著差异(10.5%vs 19.4%,P<0.01),耐药株表现出显著的TRAIL耐受现象。但联合20mmol/L氯化锂后,MTT及流式细胞术检测耐药株细胞增殖及凋亡发现H460R对rhTRAIL的敏感性显著增加。进一步研究发现死亡受体DR4和DR5的mRNA及蛋白水平升高,这可能是药物增敏的机制之一。结论:氯化锂能够增加获得性耐药细胞系H460R对TRAIL的敏感性,死亡受体表达增加是可能的增敏机制之一。
OBJECTIVE: To establish a lung cancer cell line and a lithium chloride combined with rhTRAIL cell line that acquired resistance to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) by artificial induction to understand the phenomenon and mechanism of TRAIL sensitization by lithium chloride. Methods: The human lung cancer cell line H460 was induced by low dose of rhTRAIL, and the TRAIL resistant cell line H460R was established and identified. MTT and flow cytometry were used to analyze the relationship between the drug-resistant cell lines Proliferation and apoptosis. The expression of death receptor was detected by RT-PCR and Western blot. Results: The cell viability of rhTRAIL treated parent strain H460 and drug resistant strain H460R were significantly different (P <0.01) with IC50 of 59.2ng / ml and 294.8ng / ml, respectively. There was a significant difference (P <0.01) between the average apoptosis rate of drug-resistant and parental strains treated with 60ng / ml rhTRAIL, and the resistant strains showed significant TRAIL tolerance. However, after combined with 20mmol / L lithium chloride, MTT and flow cytometry were used to detect the proliferation and apoptosis of drug-resistant cells. The sensitivity of H460R to rhTRAIL was significantly increased. Further study found that death receptors DR4 and DR5 mRNA and protein levels, which may be one of the mechanisms of drug sensitization. Conclusion: Lithium chloride can increase the sensitivity of TRAIL to H460R cell line, and the increased expression of death receptor is one of the potential mechanisms of sensitization.