摘要
目的探讨上皮细胞转化序列2(ECT2)通过调控PI3K/AKT信号通路对胰腺癌细胞恶性行为、糖酵解及TH细胞分化的影响。方法RT⁃qRCR检测人脐静脉内皮细胞系HUVEC及胰腺癌细胞中ECT2 mRNA表达,将胰腺癌Panc⁃1细胞株分为胰腺癌组(Panc⁃1细胞株正常培养)、NC组(Panc⁃1细胞株转染ECT2阴性对照)、ECT2 siRNA组(Panc⁃1细胞株转染ECT2 siRNA)、抑制剂组(Panc⁃1细胞株转染加入PI3K/AKT抑制剂LY294002),ECT2 siRNA+抑制剂组(Panc⁃1细胞株转染ECT2 siRNA加入PI3K/AKT抑制剂LY294002),采用RT⁃qRCR各组细胞中ECT2 mRNA表达;Transwell法检测各组Panc⁃1细胞侵袭能力;划痕实验检测迁移;流式细胞仪检测各组细胞IFN⁃γ及IL⁃4表达;免疫印迹分别检测糖酵解代表蛋白及PI3K/AKT表达。结果胰腺癌组、NC组、ECT2 siRNA组、抑制剂组及ECT2 siRNA+抑制剂组的ECT2 mRNA比较分别为1.00±0.00、0.95±0.03、0.41±0.08、0.65±0.05及0.20±0.04,组间比较,差异有统计学意义(F=310.700,P<0.05);胰腺癌组、NC组、ECT2 siRNA组、抑制剂组及ECT2 siRNA+抑制剂组Panc⁃1细胞侵袭数目分别为(256.30±28.36)个、(241.18±24.05)个、(155.48±17.56)个、(178.90±18.44)个及(95.15±12.10)个,组间比较,差异有统计学意义(F=59.310,P<0.01);胰腺癌组、NC组、ECT2 siRNA组、抑制剂组及ECT2 siRNA+抑制剂组Panc⁃1细胞迁移距离分别为(14.02±1.36)mm、(13.42±1.29)mm、(9.25±0.85)mm、(8.50±0.45)mm及(4.25±0.53)mm,组间比较差异有统计学意义(F=101.200,P<0.05);ECT2 siRNA组细胞IFN⁃γ升高及IL⁃4占比降低,与NC组比较,差异有统计学意义(P<0.05),与抑制剂组相比,ECT2 siRNA+抑制剂组细胞IFN⁃γ升高及IL⁃4占比降低,组间比较差异有统计学意义(P<0.05);ECT2 siRNA组细胞HIF⁃1α、GLUT1、HK⁃Ⅱ和PFK蛋白均降低,与抑制剂组无意义(P>0.05),与抑制剂组相比,ECT2 siRNA+抑制剂组细胞HIF⁃1α、GLUT1、HK⁃Ⅱ和PFK蛋白降低,组间比较差异有统计学意义(P<0.05
Objective To investigate the effects of ECT2 on malignant behavior,glycolysis and TH cell dif⁃ferentiation of pancreatic cancer cells by regulating PI3K/AKT signaling pathway.Methods ECT2 mRNA expres⁃sion in human umbilical vein endothelial cell line HUVEC and pancreatic cancer cells was detected by RT⁃QRCR.Panc⁃1 cell lines were divided into pancreatic cancer group(panc⁃1 cell line was cultured normally),NC group(PANC⁃1 cell line was transfected with ECT2 negative control)and ECT2 siRNA group(panc⁃1 cell line was trans⁃fected with ECT2 negative control SiRNA),inhibitor group(Panc⁃1 cell line transfected with PI3K/AKT inhibitor LY294002),ECT2 siRNA+inhibitor group(Panc⁃1 cell line transfected with PI3K/AKT inhibitor LY294002),ECT2 mRNA expression in rT⁃QRCR groups;The invasion ability of PANC⁃1 cells in each group was detected by Transwell assay.Migration of scratch test;The expression of IFN⁃γand IL⁃4 was detected by flow cytometry.The ex⁃pression of glycolysis representative protein and PI3K/AKT were detected by Western blot.Results ECT2 of pancre⁃atic cancer group,NC group,ECT2 siRNA group,inhibitor group and ECT2 siRNA+inhibitor group MRNA were 1.00±0.00,0.95±0.03,0.41±0.08,0.65±0.05 and 0.20±0.04,respectively,and there were differences(F=310.700,P<0.05).Pancreatic cancer group,NC group,ECT2 siRNA group,inhibitor group and ECT2 The inva⁃sion numbers of PANC⁃1 cells in the siRNA+inhibitor group were 256.30±28.36,241.18±24.05,155.48±17.56,178.90±18.44 and 95.15±12.10,respectively,and there was a difference(F=59.310,P<0.001);Pancreatic cancer group,NC group,ECT2 siRNA group,inhibitor group and ECT2 The migration distances of PANC⁃1 cells in siR⁃NA+inhibitor group were(14.02±1.36)mm,(13.42±1.29)mm,(9.25±0.85)mm,(8.50±0.45)mm and(4.25±0.53)mm,respectively(F=101.200,P<0.05).ECT2 siRNA group increased IFN⁃γand decreased IL⁃4 ratio,which was significant compared with NC group(P<0.05).Compared with inhibitor group,ECT2 siRNA+inhibitor group increased IFN⁃�
作者
鲁丁瑜
廖建
吴飞
马茜
谢非
何迎盈
LU Dingyu;LIAO Jian;WU Fei;MA Qian;XIE Fei;HE Ying-ying(Department of Oncology,Peoples Hospital of Deyang City,Deyang,Sichuan Province 618000,China;Department of Critical Care Medicine,Peoples Hospital of Deyang City,Deyang,Sichuan Province 618000,China)
出处
《解剖学研究》
CAS
2023年第4期329-336,341,共9页
Anatomy Research
基金
德阳市科技计划项目(2019SZ120和2022SCZ129)。
关键词
胰腺癌
上皮细胞转化序列2
糖酵解
TH细胞
磷脂酞肌醇3⁃激酶/蛋白激酶B
Pancreatic cancer
Epithelial cell transformation sequence 2
Glycolysis
Th cells
Phosphatidylinositol 3⁃kinase/protein kinase B(PI3K)
