摘要
目的:观察3-(5′-羟甲基-2′-呋喃)-1-苄基吲唑(YC-1)对低氧肝癌HepG-2细胞系缺氧诱导因子-1(HIF-1)稳定性和其促靶基因转录活性的调节作用,以及对细胞增殖活性的影响,探讨将HIF-1作为肿瘤治疗新靶点的可能性。方法:低氧条件下培养肝癌HepG-2细胞,应用RT-PCR和蛋白质印迹法分别检测不同YC-1作用浓度下HIF-1α及其靶基因mRNA和蛋白产物的表达。MTT检测YC-1对低氧HepG-2细胞恶性增殖力的影响。结果:低氧引起HIF-1α、VEGF和GPI mRNA表达显著增加,与常氧组比较差异有统计学意义,P<0.05。低氧条件下,YC-1以剂量依赖性方式抑制VEGF、GPI mRNA与蛋白表达,与对照组比较差异有统计学意义,P<0.05。低氧条件下,YC-1浓度升高不影响HIF-1αmRNA表达量,却以剂量依赖性方式抑制HIF-1α蛋白表达。低氧条件下,YC-1显著抑制HepG-2细胞生长,且呈现剂量依赖效应。结论:低氧状态下,YC-1抑制人肝癌HepG-2细胞系内HIF-1稳定性和转录活性,且呈剂量依赖性。YC-1抑制HepG-2细胞生长,且呈现剂量依赖效应。
OBJECTIVE： To investigate the effects of YC-1 on stability and transcriptional activity of HIF-1 in human liver cancer cells HepG-2 incubated in hypoxic conditions,and to dis cuss probable mechanism which there is. METHODS： Human liver cancer cells-HepG-2 were incubated under hypoxic culture. Semi-quantitative RT PCR and Western blotting were used to de teet the HIF let and its target genes＇ mRNA and protein levels treated with diverse concentrations of YC1. MTT assay were used to detect proliferation vitality of hypoxic HepG 2 cells. RESULTS： Comparing to the normoxic controls, for HIF-1α and its target genes, the mRNA levels highly increased in hypoxia, P〈0.05. Under hypoxic conditions, the mRNA synthesis and protein expressions of those targeted genes decreased progressively after dealt with increasing dose of YC-1 （P〈0. 05 ）, and YC-1 seemed to have no effect on the HIF-1α mRNA levels, but it de veloped a dose dependent reduction on HIF-1α protein expres sion. YC-1 inhibited the proliferation of hypoxic HepG-2 cells in a dose-dependent manner. CONCLUSIONS： YC-1 inhibits the stability and transcriptional activity of HIF-1 in a dose dependent-manner in human liver cancer cells-HepG-2 which is incubated in hypoxia. YC1 inhibits the proliferation of hypoxic HepG-2 cells in a dose-dependent manner. Our study suggests that HIF-1 might become a new potential therapeutic target for human liver cancer.
出处
《中华肿瘤防治杂志》
CAS
2009年第5期340-343,共4页
Chinese Journal of Cancer Prevention and Treatment
