摘要
目的研究乙二醛酶Ⅰ(GLO1)在胰腺癌(PC)细胞和PC干细胞(CSCs)中的表达,并探讨其对PC细胞和CSCs增殖、侵袭的影响。方法选取PC细胞系SW1990、PANC-1、BxPC3和人正常胰腺导管上皮细胞HPDE6-C7细胞,采用qRT-PCR法和Western blotting法分别检测GLO1 mRNA和蛋白表达。将呈对数生长期时的PANC-1细胞、CSCs细胞胰酶消化后铺至6孔板中,分为si-NC组和si-GLO1组,按照说明书分别将si-NC和GL01 siRNA转染试剂与脂质体2000混合后转染至si-NC组和si-GLO1组细胞中,48 h后,MTS检测细胞增殖能力,Boyden实验检测细胞侵袭能力;磁珠分选PC细胞系PANC-1中的CSCs。qRT-PCR法检测干性标志分子Nanog、OCT4、SOX2及GLO1在CSCs中的表达;si-GLO1组和si-NC组CSCs经脂质体分别转染GLO1 siRNA和si-NC 48 h后,MTS检测CSCs增殖能力,Boyden实验检测CSCs侵袭能力;Western blotting法检测干扰GLO1表达后细胞中pPIK和pAKT蛋白表达。结果qRT-PCR和Western blotting检测结果显示,GLO1 mRNA和蛋白在PC细胞系SW1990、PANC-1、BxPC3中的表达均高于人正常胰腺导管上皮细胞HPDE6-C7(P均<0.05)。干性标志分子Nanog、OCT4、SOX2在CSCs中表达增加(P均<0.05);GLO1在CSCs中的表达高于PC细胞系PANC-1(P<0.05);si-GLO1组转染GLO1 siRNA后,与si-NC组相比,PANC-1和CSCs细胞的增殖和侵袭能力均下降,细胞中pPIK和pAKT蛋白表达下降(P均<0.05)。结论GLO1在PC细胞系和CSCs中均呈高表达,干扰GLO1表达可能通过下调PI3K/AKT信号通路抑制PC细胞PANC-1和PC CSCs的增殖、侵袭。
Objective To investigate the expression of glyoxylase I(GLO1)in the pancreatic cancer(PC)cells and PC stem cells(CSCs),and to explore the effect of GLO1 on the proliferation and invasion of PC cells and CSCs.Methods PC cell line SW1990,PANC-1,BxPC3 and human normal pancreatic ductal epithelial cells HPDE6-C7 were selected.qRT-PCR and Western blotting were used to detect the mRNA and protein expression of GLO1,respectively.PANC-1 cells and CSCs in logarithmic phase were digested by trypsinase and then were laid into 6-well plates.They were divided into the si-NC group and si-GLO1 group.According to the instructions,si-NC and GL01 siRNA transfection reagents were mixed with liposome 2000 and transfected into the cells of si-NC group and si-GLO1 group,respectively.After 48 hours,MTS was used to detect cell proliferation,and Boyden test was used to detect cell invasion.CSCs in PC cell line PANC-1 were sorted by magnetic beads.The expression of dry markers Nanog,OCT4,SOX2 and GLO1 in CSCs was detected by qRT-PCR.The proliferation of CSCs was detected by MTS at 48 hours after transfection of GLO1 siRNA and si-NC by liposome,and the invasion of CSCs was detected by Boyden test.The expression of pPIK and pAKT protein was detected by Western blotting after interference with GLO1 expression.Results The qRT-PCR and Western blotting results showed that the expression of GLO1 mRNA and protein in PC cell line SW1990,PANC-1 and BxPC3 was significantly higher than that in human normal pancreatic ductal epithelial cells HPDE6-C7(P<0.05).The expression of dry markers Nanog,OCT4,and SOX2 increased in CSCs(P<0.05),the expression of GLO1 in CSCs was significantly higher than that in PC cell line PANC-1(P<0.05).After transfection with GLO1 siRNA in the si-GLO1 group,compared with si-NC group,the proliferation and invasion of PANC-1 cells and CSCs decreased,and the expression of pPIK and pAKT protein decreased(both P<0.05).Conclusions GLO1 is highly expressed in both PC cell line and CSCs.Interfering with GLO1 expression may significantly inhibit the proliferation and invasion of PANC-1 cells and CSCs by down-regulating PI3K/AKT signaling pathway.
作者
汤海涛
闫继慈
周晓琳
TANG Haitao;YAN Jici;ZHOU Xiaolin(The People’s Hospital of Liaoning Province,Shenyang 110015,China)
出处
《山东医药》
CAS
2019年第28期12-16,共5页
Shandong Medical Journal
基金
辽宁省自然科学基金计划重点项目(20170540529)
