CAJ | 학술논문

目的观察肿瘤相关钙信号传导蛋白-2(TROP-2)基因小干扰RNA(siRNA)对胃癌细胞生物学行为的影响.方法培养人胃癌MGC-803、HGC-27及BGC-823细胞株,以荧光实时定量聚合酶链反应(PCR)检测TROP-2基因mRNA表达;筛选出TROP-2表达最高者.采用TROP-2基因小干扰RNA转染胃癌细胞株,分别以荧光实时定量PCR和免疫荧光方法观察TROP-2基因mRNA和蛋白水平,然后以计数法检测细胞黏附,以划痕法观察癌细胞迁移、以Boyden方法检测癌细胞侵袭力.结果荧光实时定量PCR方法显示,3株胃癌细胞中,TROP-2均有不同程度的表达,以胃癌BGC-823细胞最高;以TROP-2 siRNA转染胃癌BGC-823细胞后,癌细胞TROP-2基因mRNA和蛋白明显下降,且呈浓度依赖性;细胞黏附结果显示,转染组细胞黏附数量明显下降;划痕试验和Boyden试验结果显示,细胞迁移和侵袭力明显下降.结论 TROP-2基囚在胃癌细胞黏附、迁移和侵袭中发挥着重要作用;以siRNA转染胃癌细胞,可抑制胃癌细胞黏附、迁移和侵袭能力.
목적 관찰종류상관개신호전도단백-2(TROP-2)기인소간우RNA(siRNA)대위암세포생물학행위적영향.방법 배양인위암MGC-803、HGC-27급BGC-823세포주,이형광실시정량취합매련반응(PCR)검측TROP-2기인mRNA표체;사선출TROP-2표체최고자.채용TROP-2기인소간우RNA전염위암세포주,분별이형광실시정량PCR화면역형광방법관찰TROP-2기인mRNA화단백수평,연후이계수법검측세포점부,이화흔법관찰암세포천이、이Boyden방법검측암세포침습력.결과 형광실시정량PCR방법현시,3주위암세포중,TROP-2균유불동정도적표체,이위암BGC-823세포최고;이TROP-2 siRNA전염위암BGC-823세포후,암세포TROP-2기인mRNA화단백명현하강,차정농도의뢰성;세포점부결과현시,전염조세포점부수량명현하강;화흔시험화Boyden시험결과현시,세포천이화침습력명현하강.결론 TROP-2기수재위암세포점부、천이화침습중발휘착중요작용;이siRNA전염위암세포,가억제위암세포점부、천이화침습능력.
Objective To study the effects of tumor-associated calcium signal transducer-2 (TROP-2) gene small interfering RNA (siRNA) on adhesion, migration, and invasion of human gastric cancer cells.Methods Real time polymerase chain reaction (PCR) was used to detect the TROP-2 mRNA expression of human gastric cancer cell lines MGC-803, HC, C-27 and BGC-823. The cells with highest expression of TROP-2 were transfected with different doses of TROP-2 siRNA. The expression of TROP-2 mRNA and protein was detected by real-time quantitative PCR and immumoflureseence method. Cell adhesion, migration,and invasion were exmined by hoyden chamber, respectively. Results Cell line BGC-823 showed the highest elevation of TROP-2 mRNA among three gastric cancer cell lines. Real-time quantitative PCR and immumoflurescence method revealed that the expression of TROP-2 mRNA and protein was reduced in a time- and dose-dependent manner ( r = 0. 935 ; r = 0. 922). The ability of adhesion, migration and invasion of BGC-823 cells treated with TROP-2 siRNA was decreased as compared with control group (P <0. 01 ). Conclusion TROP-2 gene might play an important role in adhesion, migration, and invasion of human gastric cancer cells, siRNA targeted TROP-2 could effectively inhibit adhesion, migration, and invasion of human gastric cancer cells.