Cullin1对肺腺癌A549和H1395细胞生物学特性的影响

Effects of Cullin1 on the Biological Characteristics of Lung Adenocarcinoma A549 and H1395 Cells

背景与目的Cullin1是Cullin家族中有代表性的一员,对细胞周期、转录和信号转导相关蛋白泛素化起重要作用,Cullin1与多种恶性肿瘤的发生发展有着密切的联系。本研究旨在探讨Cullin1对肺腺癌A549和H1395细胞生物功能的影响。方法实时荧光定量聚合酶链式反应(polymerase chain reaction,PCR)检测肺腺癌细胞(A549、H358、H1395、H1650)及人正常肺上皮细胞BEAS-2B中Cullin1 mRNA表达,采用siRNA技术干扰Cullin1 mRNA相对高表达的肺腺癌细胞;采用四甲基偶氮唑盐比色法(methyl thiazolyl tetrazolium assay,MTT)、流式细胞术及Transwell实验检测细胞增殖、细胞周期分布、细胞早期凋亡及侵袭和迁移能力;采用Western blot检测基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)、基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)、组织基质金属酶抑制剂-1(tissue inhibitor of metalloproteinase-1,TIMP-1)、细胞周期蛋白D1(Cyclin D1)、细胞周期蛋白E2(Cyclin E2)、p21和p27蛋白的表达水平。结果与BEAS-2B细胞相比,肺腺癌细胞中Cullin1 mRNA均呈高表达,尤其在肺腺癌A549和H1395细胞中相对表达量较高(P<0.05);干扰Cullin1后肺腺癌细胞的增殖能力受到了抑制,G1期细胞增多,S期细胞数目减少,肺腺癌细胞早期凋亡率明显升高(P<0.05);肺腺癌细胞的侵袭及迁移能力下降(P<0.05);干扰Cullin1后MMP-9、MMP-2、Cyclin D1及Cyclin E2蛋白表达量减少(P<0.05),而TIMP-1、p21和p27蛋白表达量增多(P<0.05)。结论干扰Cullin1后可抑制肺腺癌A549和H1395细胞的增殖、侵袭和迁移,Cullin1在肺腺癌中发挥促癌作用。

Background and objective Cullin1 is a representative member of the Cullin family,and it plays an important role in the ubiquitination of cell cycle,transcription and signal transduction related proteins.Cullin1 is closely related to the occurrence and development of a variety of malignant tumors.The aim of this study is to investigate the effects of Cullin1 on biological function of lung adenocarcinoma A549 and H1395 Cells.Methods The expression of Cullin1 mRNA was detected by quantitative Real-time polymerase chain reaction in lung adenocarcinoma cells(A549,H358,H1395,H1650)and human normal lung epithelial cells BEAS-2 B,siRNA technology was used to interfere with lung adenocarcinoma cells with relatively high expression of Cullin1 mRNA;cell proliferation,cell cycle distribution,early cell apoptosis,invasion and migration ability were detected by methyl thiazolyl tetrazolium assay(MTT),flow cytometry and Transwell experiment;Western blot was used to detect the expression levels of matrix metalloproteinase-2(MMP-2),matrix metalloproteinase-9(MMP-9),tissue inhibitor of metalloproteinase-1(TIMP-1),Cyclin D1,Cyclin E2,p21 and p27.Results Compared with the BEAS-2 B cell,Cullin1 mRNA was highly expressed in lung adenocarcinoma cells,especially in lung adenocarcinoma A549 and H1395 cells(P<0.05).The proliferation ability of lung adenocarcinoma cells was inhibited after interference with Cullin1,and the number of cells in G1 phase increased,the number of cells in S phase decreased,and the early apoptosis rate of lung adenocarcinoma cells is significantly increased(P<0.05);The invasion and migration ability of lung adenocarcinoma cells decreased(P<0.05).After interference with Cullin1,the protein expression of MMP-9,MMP-2,CyclinD1 and CyclinE2 decreased(P<0.05),while the expression of TIMP-1,p21 and p27 protein increased(P<0.05).Conclusion Interference with Cullin1 inhibits the proliferation,invasion and migration of lung adenocarcinoma A549 and H1395 cells,Cullin1 plays a role in promoting cancer in lung adenocarcinoma.