摘要
目的:探讨镁依赖性蛋白磷酸酶1A(PPM1A)/TGF-β1/Smads轴对胰腺癌PANC-1细胞生物学行为的影响及其可能作用机制。方法:①通过组织芯片免疫组织化学染色检测88例胰腺癌组织和14例癌旁正常胰腺组织中PPM1A的表达情况。利用qRT-PCR、免疫荧光实验检测胰腺癌细胞PANC-1、AsPC-1和BxPC-3与正常胰腺导管上皮细胞HPNE中PPM1A的表达情况。②取PANC-1细胞,采用脂质体转染法分4组,分别转染si-NC、si-PPM1A、pcDNA3.1、pcDNA3.1-PPM1A,通过CCK-8和Transwell实验检测细胞增殖、迁移和侵袭能力,Western blot法检测TGF-β1、p-Smad2/Smad2和p-Smad3/Smad3蛋白的表达情况。③取PANC-1细胞,转染si-PPM1A或pcDNA3.1-PPM1A后,再加入TGF-β1信号通路抑制剂SB431542或活化因子TGF-β1,通过CCK-8实验检测细胞增殖能力。结果:①与癌旁正常胰腺组织相比,胰腺癌组织中PPM1A蛋白表达减少(P<0.05)。②与正常胰腺导管上皮细胞HPNE相比,胰腺癌细胞PANC-1、AsPC-1和BxPC-3中PPM1A mRNA和蛋白表达减少(P<0.05)。③与si-NC组相比,si-PPM1A组细胞增殖、迁移和侵袭能力增加,TGF-β1/Smads信号通路中TGF-β1、p-Smad2/Smad2和p-Smad3/Smad3表达水平升高(P<0.05);与pcDNA3.1组相比,pcDNA3.1-PPM1A组细胞上述指标的变化相反(P<0.05)。④SB431542可逆转si-PPM1A对PANC-1细胞增殖的促进作用,TGF-β1可逆转pcDNA3.1-PPM1A对PANC-1细胞增殖的抑制作用(P<0.05)。结论:PPM1A可能通过抑制TGF-β1/Smads信号通路抑制胰腺癌PANC-1细胞的增殖、迁移和侵袭能力。
Aim:To investigate the effects of protein phosphatase magnesium dependent 1A(PPM1A)/TGF-β1/Smads axis on the biological behavior of pancreatic cancer PANC-1 cells and its possible mechanism.Methods:The expression of PPM1A in 88 cases of pancreatic cancer and 14 cases of normal adjacent pancreatic tissue from pancreatic cancer patients was detected by immunohistochemical staining with tissue microarray.The expression of PPM1A was detected by qRT-PCR and immunofluorescence assay in pancreatic cancer PANC-1,AsPC-1,BxPC-3 cells and normal pancreatic duct epithelial HPNE cells.PANC-1 cells were transfected with si-NC,si-PPM1A,pcDNA3.1 and pcDNA3.1-PPM1A,respectively,by liposome transfection method,CCK-8 and Transwell assays were used to detect cell proliferation,migration and invasion abilities,and Western blot was used to detect the expressions of TGF-β1,p-Smad2/Smad2 and p-Smad3/Smad3.CCK-8 was performed to detect whether TGF-β1/Smads signal pathway inhibitor SB431542 or activation factor TGF-β1 could reverse the promotional or inhibitory effects of si-PPM1A or pcDNA3.1-PPM1A on the proliferation in PANC-1 cells.Results:The protein expression of PPM1A in pancreatic cancer tissue was significantly lower than that in normal adjacent pancreatic tissue(P<0.05).The mRNA and protein expressions of PPM1A in pancreatic cancer PANC-1,AsPC-1 and BxPC-3 cells were significantly lower than those in normal pancreatic duct epithelial HPNE cells(P<0.05).The proliferation,migration and invasion abilities of PANC-1 cells,as well as the protein expression levels of TGF-β1,p-Smad2/Smad2 and p-Smad3/Smad3 in TGF-β1/Smads signaling pathway in si-PPM1A group were significantly higher compared with si-NC group,which were lower in pcDNA3.1-PPM1A group compared with pcDNA3.1 group(P<0.05).SB431542 or TGF-β1 reversed the promotional or inhibitory effects of si-PPM1A or pcDNA3.1-PPM1A on the proliferation of PANC-1 cells(P<0.05).Conclusion:PPM1A may inhibit the proliferation,migration and invasion of pancreatic cancer PANC-1 cells by i
作者
张荣花
崔笑妍
周静
王梅梅
熊亚南
于笑涵
刘志勇
章广玲
ZHANG Ronghua;CUI Xiaoyan;ZHOU Jing;WANG Meimei;XIONG Yanan;YU Xiaohan;LIU Zhiyong;ZHANG Guangling(School of Basic Medical Sciences,North China University of Science and Technology,Hebei Provincial Key Laboratory for Chronic Diseases,Tangshan,Hebei 063210;Clinical Medicine School,North China University of Science and Technology,Hebei Provincial Key Laboratory of Medical-Industrial Integration Precision Medicine,Tangshan,Hebei 063000)
出处
《郑州大学学报（医学版）》
CAS
北大核心
2024年第4期463-469,共7页
Journal of Zhengzhou University(Medical Sciences)
基金
河北省自然科学基金项目(H2024209077,H2023209047)
河北省人力资源和社会保障厅项目(C20210340)
河北省重点研发计划项目(213777115D)
河北省财政厅项目(冀财预复[2020]397号)。
