摘要
目的:探讨索拉非尼能否逆转人肝癌细胞株HepG2/GEM的耐药性及可能机制。方法:以药物大剂量冲击法建立耐药细胞模型HepG2/GEM,MTT法检测索拉非尼对HepG2、HepG2/GEM的抑制率及索拉非尼处理前后2株细胞对GEM敏感性的变化,RT-PCR法检测MDR-1及ERK、VEGFR-2mRNA水平的变化,蛋白质印迹法检测P-gp、VEG-FR-2蛋白水平的变化。结果:建立了人肝癌HepG2/GEM耐药细胞模型,耐药倍数11.63倍。经1mg/L的索拉非尼处理后,GEM对HepG2/GEM细胞株的IC50明显下降,逆转倍数为3.08倍,相对逆转率为73.87%,P=0.019。1mg/L的索拉非尼能够降低HepG2/GEM细胞MDR-1、ERK、VEGFR-2mRNA表达(P=0.033),并可明显抑制P-gp、VEGFR-2蛋白的表达,P=0.007。结论:经GEM大剂量间断冲击后,HepG2细胞株对GEM产生了耐药性。索拉非尼具有体外部分逆转HepG2/GEM细胞株耐药性的作用。可能与抑制VEGFR-2、ERK1/2信号传导通路,从而下调MDR-1有关。
OBJECTIVE：To explore whether sorafenib can reverse the resistance to gemcitabine（GEM）of human hepatocellular carcinoma HepG2/GEM cell line in vitro and its possible mechanism.METHODS：The human hepatocellular carcinoma GEM-resistant cell subline HepG2/GEM was induced after treated with GEM by high dose pulse treatment selection.The MTT assay was used to detect the inhibition ratio of sorafenib to HepG2/GEM and HepG2 cell lines and the drug sensitivity of HepG2/GEM and HepG2 cell lines to GEM which were exposed to sorafenib or not.The RT-PCR assay was applied to determine the changes of MDR1,ERK and VEGFR-2 mRNA levels,and the Western blot assay was applied to test the changes of the expression levels of P-gp and VEGFR-2.RESULTS：The human hepatocellular carcinoma GEM-resistant cell subline named as HepG2/GEM cell line was obtained by high dose pulse treatment selection successfully.The final level of resistace to GEM of HepG2/GEM was 11.63-fold compared to parental HepG2 cells.The IC50 value of GEM to HepG2/GEM was decreased obviously after treated with sorafenib in 1 mg/L compared to HepG2/GEM cells not treated with sorafenib.The final reversal levels of resistace to GEM of HepG2/GEM was 3.08-fold.The relative reversal rate was 73.87% （P=0.019）.The MDR-1,ERK,VEGFR-2 mRNA level of HepG2/GEM cell line were reduced compared to its prior levels （P=0.033） and the expression levels of P-gp and VEGFR-2 were significantly suppressed after treated with sorafenib in 1 mg/L （P=0.007）.CONCLUSIONS：The HepG2 cells can be induced in vitro to be the acquired GEM-resistant lines by high dose pulse treatment selection.Sofafenib can reverse the drug-resistance to GEM of human hepatoma carcinoma cells,which is associated with decreasing the expressions of MDR1/P-gp,VEGFR-2 and ERK1/2.
出处
《中华肿瘤防治杂志》
CAS
2010年第15期1160-1163,共4页
Chinese Journal of Cancer Prevention and Treatment
关键词
癌
肝细胞
抗药性
肿瘤
抗肿瘤药
carcinoma
hepatocellular
drug resistance
neoplasms
antineoplastic agents
