PIAS1通过调控Sonic hedgehog信号通路使胰腺癌细胞SW1990获得对吉西他滨的耐药性

目的:研究吉西他滨耐药性胰腺癌细胞中Sonic hedgehog(Shh)信号通路异常激活的机制?方法:通过间歇梯度倍增法筛选出对吉西他滨产生耐药性的胰腺癌细胞SW1990-GEM,荧光实时定量PCR和蛋白质印迹法检测耐药细胞中PIAS1的表达;利用RNA干扰技术构建稳定低表达PIAS1的细胞株SW1990-GEM-shPIAS1,从增殖速度?克隆形成能力及裸鼠皮下成瘤能力等方面考察PIAS1?Shh信号通路与胰腺癌耐药性的相关性?结果:耐药株SW1990-GEM中PIAS1基因表达量明显升高;PIAS1的高度表达正向激活了Shh信号通路的活性,使得细胞获得耐药能力;人为降低PIAS1的高表达后,耐药株的耐药能力同时被减弱?结论:PIAS1是使耐药株获得耐药性的关键因子,PIAS1通过正向调控Shh信号通路使其获得耐药能力?

PIAS1 promotes acquired gemcitabine resistance in pancreatice cancer cell SW1990 through regulating Sonic hedgehog signaling

Objective:To investigate the mechanism of aberrant Sonic hedgehog (Shh) signaling which contributes to acquired resistance to gemcitabine of pancreatic cancer cells SW1990. Methods:Gemcitabine-resistant pancreatic cancer cells SW1990-GEM were induced by increasing drug dosage intermittently. Quantitative real-time PCR and Western blot assay were used to evaluate the relative expression level of PIAS1 in SW1990 cells and SW1990-GEM cells. The ability of PIAS1 promoting acquired resistance to gemcitabine through up-regulating Shh signaling was evaluated by cell proliferation,soft agar colony formation assay and nude mice tumor transplantation assay. Results:SW1990-GEM (resistant strains) showed a high expression level of PIAS1. The overexpression of PIAS1 was involved in the increased cell proliferation and tumor formation ability of SW1990-GEM cells. Knockdown of PIAS1 in SW1990-GEM reduced the activity of Shh signaling thereby decreasing the drug resistance ability of this strain. Conclusion:PIAS1 is the key factor for acquired resistance to gemcitabine in pancreatice cancer cell SW1990 by up-regulating Shh signaling.