成纤维生长因子2在胰腺癌中的表达及对胰腺癌细胞侵袭与转移的作用和机制

Expression of fibroblast growth factor 2 in pancreatic ductal adenocarcinoma tissues and its effects on invasion and metastasis of PANC-1

目的探讨成纤维生长因子2(FGF2)在胰腺癌中的表达及对胰腺癌PANC-1细胞侵袭转移的作用和机制。方法收集62例患者的胰腺导管细胞癌(PDAC)组织和癌旁正常胰腺(NP)组织,免疫组化法检测癌组织和癌旁正常胰腺组织中FGF2的表达。将RPMI 1640培养的人胰腺癌细胞PANC-1分为4组:对照组、FGF2组、FGFR2+AZD4547组和FGFR2+LY294002组。Western blot及RT-PCR分别检测palladin和Akt的蛋白及mRNA表达。Transwell小室细胞侵袭活性实验和细胞划痕实验检测各组PANC-1细胞侵袭和迁移的能力。结果免疫组化显示,胰腺癌组织中FGF2表达率81%(50/62)高于癌旁正常组织的10%(2/20,P<0.05),且胰腺癌中FGF2表达与肿瘤的分化程度、临床TNM分期、淋巴结转移和远处转移有关(P<0.05),与患者性别、年龄和肿瘤部位无关(P>0.05)。Western blot测定显示:FGF2组palladin和p-Akt的蛋白表达高于其他组(P<0.05)。RT-PCR结果显示:FGF2组palladin和p-Akt的mRNA表达高于其他组(P<0.05)。Transwell小室侵袭活性实验结果表明:FGF2组侵袭细胞数高于其他组(P<0.05),FGFR2+AZD4547组和FGFR2+LY294002组侵袭细胞数低于对照组和FGF2组(P<0.05)。细胞划痕实验结果表明:FGF2组细胞迁移率高于其他组(P<0.05),FGFR2+AZD4547组和FGFR2+LY294002组细胞迁移率低于对照组和FGF2组(P<0.05)。结论 FGF2在胰腺癌组织中高表达,与胰腺癌的侵袭和转移相关,其机制可能是FGF2与其受体FGFR2结合后活化PI3K/Akt信号通路,激活并提高palladin的表达,促进细胞的运动,进而促进肿瘤细胞的侵袭与迁移活动。

Objective To investigate the expression of fibroblast growth factor 2（ FGF2） in pancreatic ductal adenocarcinoma（ PDAC）tissues and its role in the invasion and metastasis of PANC-1. Methods Immunohistochemistry assay was performed to detect the expression of FGF2 in 62 cases of PDAC tissues and corresponding adjacent normal pancreas（NP） tissues. The human pancreatic cancer PANC-1 cells were cultured in RPMI 1640 culture medium. The cells were randomly divided into 4 groups：control group,FGF2 group,FGF2 ＋ AZD4547 group and FGF2 ＋ LY294002 group. Western blot analysis and RT-PCR were performed respectively to detect the expression of palladin and Akt in PANC-1 cells. Transwell chamber assay and wound healing assay were applied to determine the invasive activity and the migration of PANC-1 cells respectively. Results Immunohistochemical staining showed that FGF2 was highly expressed in PDAC tissues. The rate of FGF2 expression was 81%（50/62） in PDAC tissues,significantly higher than that of NP tissues（10. 0%,2/20）（P 〈 0. 05）. And FGF2 expression was correlated with the degree of tumor differentiation,clinical TNM classification,lymph node metastasis and distant metastasis in pancreatic tissues（ P 〈 0. 05）,but not related with patient’s sex,age and tumor location（P 〉 0. 05）. Western blot analysis revealed that the relative expression of FGF2 and p-Akt in FGF2 group was higher than that in the other groups（ P 〈 0. 05）. RT-PCR results showed that the relative expression of FGF2 mRNA and Akt mRNA in FGF2 group was higher than that in the other groups（ P 〈 0. 05）. Transwell chamber assay showed that the invasion cells numbers in FGF2 group were higher than that in other groups（ P 〈 0. 05）. The invasion cells numbers in AZD4547 group and LY294002 group were lower than that in control group and FGF2 group（ P 〈 0. 05）. The wound healing assay showed that the migration rate in FGF2 group was higher than that in the other groups（ P 〈 0. 05）. The migration rates in AZD4547 group and LY294002 group were lower than that in control group and FGF2 group（ P 〈 0. 05）. Conclusion FGF2 is highly expressed in PDAC tissues,and is related to invasion and metastasis of PDAC.FGF2 could enhance the invasion and migration of pancreatic tumor cells by binding to its receptor FGFR2 for activating PI3K/Akt signaling pathway and activating and highly expressing palladin.