PSC经HGF/c-Met/survivin信号通路对胰腺癌细胞侵袭转移影响

EFFECT OF PANCREATIC STELLATE CELL ON INVASION AND METASTASIS OF PANCREATIC CANCER CELLS VIA THE HGF/C-MET/SURVIVIN SIGNALING PATHWAY

目的探讨胰腺星状细胞(PSC)对胰腺癌细胞侵袭转移的作用及其机制。方法收集PSC上清液(PSC-CM),采用ELISA法检测其中肝细胞生长因子(HGF)含量。实验分为5组:对照组(胰腺癌细胞株Panc-1)、PSC-CM组(5mL的PSC-CM孵育Panc-1细胞24h)、PSC-CM+HGF-抗体组(5 mL的PSC-CM+1μg的HGF-抗体孵育Panc-1细胞24h)、PSC-CM+SU11274组(5 mL的PSC-CM+0.25μg的SU11274孵育Panc-1细胞24h)、PSC-CM+siRNA-survivin组(Panc-1细胞转染siRNA-survivin后,用5 mL的PSC-CM孵育24h)。构建Transwell模型测定各组Panc-1细胞的转移能力,采用Western Blot法检测各组E-cadherin、Vimentin、Survivin蛋白的表达水平。结果 PSC-CM中24、48h的HGF浓度分别为(1 423.1±122.9)、(1 869.6±84.8)ng/L。PSC-CM组较对照组、PSC-CM+HGF-抗体组、PSC-CM+SU11274组、PSC-CM+siRNA-survivin组细胞转移能力明显增强,E-cadherin蛋白表达水平明显下降(F=79.70,P<0.01),Vimentin、Survivin蛋白表达水平升高(F=30.41、58.27,P<0.01)。结论 PSC可能通过HGF/c-Met/survivin信号通路促进胰腺癌细胞的转移。

Objective To investigate the effect of pancreatic stellate cells （PSCs） on the invasion and metastasis of pan- creatic cancer cells and related mechanisms. Methods The supernatant of PSCs （PSC-CM） was collected and ELISA was used to measure the content of hepatocyte growth factor （HGF）. There were five experimental groups, i.e., control group （pancreatic cancer cell line Panc-l）, PSC-CM group （Panc-1 ceils treated with 5 mL PSC-CM for 24 h）, PSC-CMq-HGF-antibody group （Panc-] ceils treated with 5 mL PSC-CM＋ 1 μg HGF-antibody for 24 h）, PSC-CM＋ SUl1274 group （Panc-1 ceils treated with 5 mL PSC-CM＋0.25μg SUl1274 for 24 h）, and PSC-CM＋siRNA-survivin group （Panc-1 cells trans{ected with siRNA-survivin and then treated with 5 mL PSC-CM for 24 h）. A Transwell model was established to measure the transfer ability of Panc-1 cells, and Western Blot was used to measure the protein expression of E-cadherin, Vimentin, and Survivin. Results The concentra- tion of HGF in PSC-CM was （1 423.1±122.9） ng/L at 24 h and （1 869.6±84.8） ng/L at 48 h. Compared with the other four groups, the PSC-CM group had a significantly stronger transfer ability, a significant reduction in the protein expression of E-cad- herin （F=79.70,P〈0.01）, and significant increases in the protein expression of Vimentin and Survivin （F=30.41,58.27;P〈 0.01）. Conclusion PSCs may promote the metastasis of pancreatic cancer cells via the HGF/c-Met/survivin signaling pathway.