中华肝胆外科杂志
中華肝膽外科雜誌
중화간담외과잡지
CHINESE JOURNAL OF HEPATOBILIARY SURGERY
2013年
12期
938-942
,共5页
江建新%赖智文%刘勇%孙城谊
江建新%賴智文%劉勇%孫城誼
강건신%뢰지문%류용%손성의
胰腺肿瘤%基因,14-3-3 sigma%细胞增殖
胰腺腫瘤%基因,14-3-3 sigma%細胞增殖
이선종류%기인,14-3-3 sigma%세포증식
Pancreatic neoplasms%Gene,14-3-3 sigma%Cell proliferation
目的 探讨14-3-3 sigma(14-3-3σ)基因对胰腺癌细胞株PANC-1增殖能力的影响.方法 采用免疫组化方法检测46例胰腺癌标本及9例正常胰腺组织标本中的14-3-3σ的表达.采用RT-qPCR和蛋白印迹检测胰腺癌细胞株BxPC3、PANC-1、AsPC-1、SW1990、MiaPaCa-2、CFPAC-1和Capan-1中14-3-3σ的mRNA和蛋白表达水平;构建人14-3-3σ基因真核表达质粒pEGFP-14-3-3σ,蛋白印迹、RT-qPCR验证pEGFP-14-3-3σ的表达,MTS法检测细胞增殖活性.结果 14-3-3σ蛋白在胰腺癌组织中较正常胰腺组织高表达.14-3-3σ的mRNA和蛋白在BxPC3、AsPC-1、SW1990和CFPAC-1高表达,在PANC-1、Capan-1和MiaPaCa-2中低表达.RT-qPCR、蛋白印迹证实pEGFP-14-3-3σ构建成功.MTS法检测结果显示转染pEGFP-14-3-3σ后的PANC-1细胞较对照组增殖能力增加.结论 14-3-3σ在胰腺癌中高表达,增强胰腺癌细胞株PANC-1的增殖能力;14-3-3σ有可能成为胰腺癌基因治疗的靶点.
目的 探討14-3-3 sigma(14-3-3σ)基因對胰腺癌細胞株PANC-1增殖能力的影響.方法 採用免疫組化方法檢測46例胰腺癌標本及9例正常胰腺組織標本中的14-3-3σ的錶達.採用RT-qPCR和蛋白印跡檢測胰腺癌細胞株BxPC3、PANC-1、AsPC-1、SW1990、MiaPaCa-2、CFPAC-1和Capan-1中14-3-3σ的mRNA和蛋白錶達水平;構建人14-3-3σ基因真覈錶達質粒pEGFP-14-3-3σ,蛋白印跡、RT-qPCR驗證pEGFP-14-3-3σ的錶達,MTS法檢測細胞增殖活性.結果 14-3-3σ蛋白在胰腺癌組織中較正常胰腺組織高錶達.14-3-3σ的mRNA和蛋白在BxPC3、AsPC-1、SW1990和CFPAC-1高錶達,在PANC-1、Capan-1和MiaPaCa-2中低錶達.RT-qPCR、蛋白印跡證實pEGFP-14-3-3σ構建成功.MTS法檢測結果顯示轉染pEGFP-14-3-3σ後的PANC-1細胞較對照組增殖能力增加.結論 14-3-3σ在胰腺癌中高錶達,增彊胰腺癌細胞株PANC-1的增殖能力;14-3-3σ有可能成為胰腺癌基因治療的靶點.
목적 탐토14-3-3 sigma(14-3-3σ)기인대이선암세포주PANC-1증식능력적영향.방법 채용면역조화방법검측46례이선암표본급9례정상이선조직표본중적14-3-3σ적표체.채용RT-qPCR화단백인적검측이선암세포주BxPC3、PANC-1、AsPC-1、SW1990、MiaPaCa-2、CFPAC-1화Capan-1중14-3-3σ적mRNA화단백표체수평;구건인14-3-3σ기인진핵표체질립pEGFP-14-3-3σ,단백인적、RT-qPCR험증pEGFP-14-3-3σ적표체,MTS법검측세포증식활성.결과 14-3-3σ단백재이선암조직중교정상이선조직고표체.14-3-3σ적mRNA화단백재BxPC3、AsPC-1、SW1990화CFPAC-1고표체,재PANC-1、Capan-1화MiaPaCa-2중저표체.RT-qPCR、단백인적증실pEGFP-14-3-3σ구건성공.MTS법검측결과현시전염pEGFP-14-3-3σ후적PANC-1세포교대조조증식능력증가.결론 14-3-3σ재이선암중고표체,증강이선암세포주PANC-1적증식능력;14-3-3σ유가능성위이선암기인치료적파점.
Objective To explore the overexpression of the 14-3-3 sigma (14-3-3σ) gene effects proliferation in pancreatic cancer cells.Methods The 14-3-3σ protein level in 46 pancreatic cancers and 9 normal pancreases specimens were analyzed by immunohistochemistry.The mRNA and protein expression of 14-3-3σ in pancreatic carcinoma cell lines (BxPC3,CAPAN-1,PANC-1,AsPC-1,SW1990,MiaPaCa-2 and CFPAC-1) were detected by RT-qPCR and Western blot respectively.A recombinant plasmid of pEGFP-14-3-3σ was constructed and transfected into the pancreatic cancer cell PANC-1 by liposome,and the expression of 14-3-3σ was detected by Western blot and real time fluorescence quantitative PCR.Cell proliferation activity was determined by MTS assay.Results The 14-3-3σ protein level was higher in pancreatic cancer tissue than in normal pancreatic tissue.mRNA and protein expression of 14-3-3σ was highest in BxPC3,high in AsPC-1,SW1990 and CFPAC-1,low in PANC-1 and Capan-1,and lowest in MiaPaCa-2.The successfully constructed pEGFP-14-3-3σ was confirmed by RT-qPCR and Western blot.MTS assay showed cell proliferation activity was significantly enhanced by overexpression of the 14-3-3σ gene compared to negative and blank control cells.Conclusion The expression of 14-3-3σ was higher in pancreatic cancer compared with normal pancre atic tissue,and the 14-3-3σ gene enhanced the cell proliferation activity of PANC-1.Therefore,14-3-3σ may play an important role in pancreatic cancer development.
