胰腺星状细胞通过HGF/c-Met信号通路调控胰腺癌的侵袭转移

摘要

Objective To investigate the regulation mechanism of pancreatic stellate cells (PSCs) on invasion and metastasis of human pancreatic cancer through the pathway of HGF/c-Met.Methods Hepatocyte growth factor (HGF) level in PSCs supernatant was detected.PSCs supernatant,PSCs supernatant plus antiHGF antibody,PSCs supernatant plus c-Met specific inhibitor PHA 665752 were used to treat human pancreatic cancer AsPC-1 cells,and untreated cells served as controls.MTT assay was applied to detect cell proliferation.Transwell chamber migration assay was employed to detect cell migration.In vitro invasion assay was used to determine cell invasion.Results The level of HGF in PSCs supernatant was (4 213 ± 543) ng/L.A490 value for cell proliferation in PSCs supernatant,PSCs supernatant + anti-HGF,PSCs supernatant + c-Met inhibitor and control group were 0.628 ± 0.030,0.324 ± 0.021,0.347 ± 0.054 and 0.405 ± 0.008.The number of penetrating cells per 400 high power field was 123.3 ± 6.8,62.4 ± 6.9,58.1 t 2.2 and 36.6 ± 4.8,respectively.The number of invasive cells per 400 high power field was 70.0 ± 2.3,42.5 ± 4.6,42.7 ± 2.8 and 36.4 ± 3.5.The proliferation,migration and invasion of pancreatic cancer AsPC-1 cells in PSCssupernatant group were significantly higher than those in the control group (all P ＜ 0.01).The proliferation,migration and invasion of pancreatic cancer AsPC-1 cells in PSCs supematant + anti-HGF and PSCs supernatant + c-Met inhibitor group were significantly lower than those in PSCs supernatant group(P ＜ 0.01),but there was no difference between the PSCs supernatant + anti-HGF group and PSCs supernatant + c-Met inhibitor group (all P ＞ 0.05).Conclusions PSCs can promote cell proliferation,migration and invasion of pancreatic cancer AsPC-1 cells via regulating HGF/c-Met pathway.%目的 探讨胰腺星状细胞(PSCs)调控胰腺癌的迁移和侵袭转移机制.方法 检测PSCs培养上清液中肝细胞生长因子(HGF)蛋白的含量.应用PSCs上清液、PSCs上清液加HGF中和抗体或加c-Met特异性抑制剂PHA-665752分别处理人胰腺癌细胞株AsPC-1细胞,以不加PSCs上清液细胞作为对照组,采用MTT法检测AsPC-1细胞的增殖,Transwell小室检测细胞迁移能力,体外侵袭实验观察细胞侵袭能力.结果 PSCs上清液中HGF蛋白含量为(4 213 ±543) ng/L.PSCs上清液组、PSCs上清液+ HGF中和抗体组、PSCs上清液+c-Met抑制剂组及对照组细胞增殖的A490值分别为0.628±0.030、0.324±0.021、0.347±0.054及0.405±0.008,穿膜细胞数分别为(123.3±6.8)、(62.4±6.9)、(58.1±2.2)、(36.6±4.8)/400倍视野,侵袭细胞数分别为(70.0±2.3)、(42.5±4.6)、(42.7±2.8)、(36.4±3.5)/400倍视野.PSCs上清液组细胞增殖、穿膜及侵袭能力较对照组显著升高,PSCs上清液+HGF中和抗体组及PSCs上清液+c-Met抑制剂组细胞增殖、穿膜及侵袭能力较PSCs上清液组显著下降,差异均有统计学意义(P值均＜0.01),而PSCs上清液+HGF中和抗体组与PSCs上清液+c-Met抑制剂组之间的细胞增殖、穿膜及侵袭能力差异无统计学意义(P值均＞0.05).结论 PSCs可能是通过HGF/c-Met信号通路从而调控胰腺癌细胞的增殖、迁移和侵袭.