干扰PIM3的表达对人胰腺癌AsPC-1细胞增殖和凋亡的影响及作用机制研究  

Effect of PIM3 interference on the proliferation and apoptosis of human pancreatic cancer AsPC 1 cells and its underlying mechanism

作　　者：陈慧 常瑛 刘振国 CHEN Hui;CHANG Ying;LIU Zhenguo(Department of Clinical Pharmacy Room of Pharmacy,Northwest Women and Children’s Hospital,Xi’an 710000,Shaanxi,China)

机构地区：[1]西北妇女儿童医院药剂科临床药学室,西安710000

出　　处：《癌症进展》2021年第18期1863-1867,共5页Oncology Progress

摘　　要：目的探讨干扰PIM3的表达对人胰腺癌AsPC-1细胞增殖和凋亡的影响及作用机制。方法将阴性对照[scramble小干扰RNA(siRNA)]和干扰PIM3(PIM3 siRNA)分别转染至人胰腺癌AsPC-1细胞,分别作为scramble-siRNA组和PIM3-siRNA组,以1×磷酸盐缓冲液(PBS)作为溶剂对照。采用实时荧光定量聚合酶链反应(RT-PCR)检测人胰腺癌AsPC-1细胞PIM3、B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(BAX)、Ki-67、caspase 3 mRNA的相对表达量,蛋白质印迹法(Westernblot)检测PIM3、Bcl-2、BAX、caspase 3蛋白的相对表达量;四甲基偶氮唑蓝(MTT)检测干扰PIM3的表达对人胰腺癌AsPC-1细胞增殖能力的影响,流式细胞仪检测干扰PIM3的表达对细胞凋亡能力的影响。探讨干扰PIM3的表达对荷瘤小鼠人胰腺癌AsPC-1细胞的影响。结果转染后,PIM3-siRNA组细胞PIM3 mRNA和PIM3蛋白的相对表达量均明显低于溶剂对照组和scramble-siRNA组(P﹤0.01)。干扰PIM3的表达可明显抑制人胰腺癌AsPC-1细胞增殖,促进其凋亡。干扰PIM3的表达可下调人胰腺癌AsPC-1细胞中Ki-67、Bcl-2基因的表达,上调BAX、caspase 3基因的表达;且干扰PIM3的表达可下调Bcl-2蛋白的表达,上调BAX、caspase 3蛋白的表达。荷瘤小鼠经PIM3 siRNA治疗后,小鼠肿瘤质量和体积均明显下降。结论PIM3 siRNA可通过干扰PIM3的表达,诱导人胰腺癌AsPC-1细胞的凋亡,在体内体外均可抑制胰腺癌AsPC-1细胞的生长。Objective To investigate the effect of PIM3 interference on the proliferation and apoptosis of human pancreatic cancer AsPC-1 cells and its underlying mechanism.Method The negative control[scramble small interfering RNA(siRNA)]and PIM3 expression interference(PIM3 siRNA)were transfected into human pancreatic cancer AsPC-1 cells,respectively,as the scramble-siRNA group and PIM3-siRNA group,with 1×phosphate buffered saline(PBS)as the solvent contrast.The relative expression levels of human pancreatic cancer AsPC-1 cells PIM3,B cell lymphoma/leukemia-2(Bcl-2),Bcl-2-associated X protein(BAX),Ki-67,caspase 3 mRNA using reverse transcription-polymerase chain reaction(RT-PCR)method for detection.Western blot detection of the relative expression of PIM3,Bcl-2,BAX,and caspase 3 proteins.The effect of PIM3 interference on the proliferation of human pancreatic cancer AsPC-1 cells was detected by methyl thiazolyl terazolium(MTT)assay.The effect of interfering with the expression of PIM3 on the apoptotic ability of human pancreatic cancer AsPC-1 cells was detected by flow cytometry.The effect of interfering with the expression of PIM3 on human pancreatic cancer AsPC-1 cells in tumor-bearing mice was also investigated.Result After PIM3 siRNA was transfected into human pancreatic cancer AsPC-1 cells,the relative expressions of PIM3 mRNA and PIM3 protein in the PIM3-siRNA group were significantly lower than those in the solvent contrast and the scramble-siRNA group(P<0.01).PIM3 interference could significantly inhibit the proliferation of human pancreatic cancer AsPC-1 cells and promote their apoptosis.PIM3 interference could down-regulate the expression of Ki-67 and Bcl-2 genes in human pancreatic cancer AsPC-1 cells,and up-regulate the expression of BAX and caspase 3 genes.And interference with the expression of PIM3 could down-regulate the expression of Bcl-2 protein,and up-regulate the expression of BAX and caspase 3 proteins.After the tumor-bearing mice were treated with PIM3 siRNA,the mass and volume of the tumors in the

关 键 词：胰腺癌 小干扰RNA AsPC-1细胞 增殖 凋亡 

分 类 号：R735.9[医药卫生—肿瘤]