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)对乳腺癌细胞黏附和侵袭力的影响.方法培养人乳腺癌Bcap-37、LCC1、MCF-7、MDA-MB-231、MDA-MB-435、MDA-MB-468及ZR75-1细胞株,以荧光实时定量聚合酶链反应(PCR)方法检测TROP-2基因mRNA表达；筛选出TROP-2表达最高者.采用TROP4基因siRNA转染乳腺癌细胞,分别以荧光实时定量RT-PCR和免疫荧光方法观察TROP-2基因mRNA和蛋白水平,然后以噻唑蓝(MTT)比色法检测细胞黏附性,以Transwell方法检测癌细胞侵袭能力.结果人乳腺癌Bcap-37、LCC1、MCF-7、MDA-MB-231、MDA-MB-35、MDA-MB-468及ZR75-1细胞株TROP-2 mRNA分别是1.362±0.057、2.207±0.056、2.997±0.052、0.136±0.045、0.122±0.025、0.194±0.028和2.706±0.039,以MCF-7细胞最高;以TROP-2 siRNA转染乳腺癌MCF-7细胞后,癌细胞TROP-2基因mRNA和蛋白水平明显下降,且呈浓度依赖性；黏附实验结果显示,5、10、20 nmol/L siRNA组黏附率分别为(52.9±2.5)％、(25.6±2.3)％、(12.8±2.2)％(P＜0.01)；Transwell实验结果显示,5、10和20 mol/L siRNA组穿过滤膜的细胞分别为78±17、39±15、19±16,而对照组分别为136±25、139±21(P＜0.01).结论 TROP-2基因在乳腺癌细胞黏附和侵袭中发挥着重要作用；以siRNA转染乳腺癌细胞,可抑制乳腺癌细胞黏附和侵袭能力.
목적 관찰종류상관개신호전도단백-2(TROP-2)기인소간우RNA (siRNA)대유선암세포점부화침습력적영향.방법 배양인유선암Bcap-37、LCC1、MCF-7、MDA-MB-231、MDA-MB-435、MDA-MB-468급ZR75-1세포주,이형광실시정량취합매련반응(PCR)방법검측TROP-2기인mRNA표체；사선출TROP-2표체최고자.채용TROP4기인siRNA전염유선암세포,분별이형광실시정량RT-PCR화면역형광방법관찰TROP-2기인mRNA화단백수평,연후이새서람(MTT)비색법검측세포점부성,이Transwell방법검측암세포침습능력.결과 인유선암Bcap-37、LCC1、MCF-7、MDA-MB-231、MDA-MB-35、MDA-MB-468급ZR75-1세포주TROP-2 mRNA분별시1.362±0.057、2.207±0.056、2.997±0.052、0.136±0.045、0.122±0.025、0.194±0.028화2.706±0.039,이MCF-7세포최고;이TROP-2 siRNA전염유선암MCF-7세포후,암세포TROP-2기인mRNA화단백수평명현하강,차정농도의뢰성；점부실험결과현시,5、10、20 nmol/L siRNA조점부솔분별위(52.9±2.5)％、(25.6±2.3)％、(12.8±2.2)％(P＜0.01)；Transwell실험결과현시,5、10화20 mol/L siRNA조천과려막적세포분별위78±17、39±15、19±16,이대조조분별위136±25、139±21(P＜0.01).결론 TROP-2기인재유선암세포점부화침습중발휘착중요작용；이siRNA전염유선암세포,가억제유선암세포점부화침습능력.
Objective To study the effects of tumor-associated calcium signal transducer-2 (TROP-2) gene small interfering RNA (siRNA) on adhesion and invasion of human breast cancer cells.Methods Real time PCR was used to detect the TROP-2 mRNA of seven human breast cancer cell lines:Bcap-37,LCC1,MCF-7,MDA-MB-231,MDA-MB-435,MDA-MB-468,and ZR75-1.The cell line with hifhest TROP-2 expression was transfected with different doses of TROP-2 siRNA.The expression of TROP-2 mRNA and protein was detected by real-time quantitative polymerase chain reaction (PCR) and immumoflurescence method.The cell adhesion was evaluated by methyl thiazol tetrazolium (MTT) assay,and invasion was exmined by boyden chamber,respectively.Results The TROP-2 mRNA in Bcap-37,LCC1,MCF-7,MDA-MB-231,MDA-MB-435,MDA-MB-468 and ZR75-1 cell lines was 1.362 ±0.057,2.207 ± 0.056,2.997 ± 0.052,0.136 ± 0.045,0.122 ± 0.025,0.194 ± 0.028 and 2.706 ± 0.039 respectively,and MCF-7 showed the highest elevation of TROP-2 mRNA.The 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 (P ＜ 0.01 ;P ＜ 0.01 ).The adhesive rate in siRNA groups (5,10 and 20 nmol/L) was (52.9±2.5)％,(25.6±2.3)％ and (12.8±2.2)％ (P＜0.01),respectively.The transwell results showed that the invasion cells were 78 ± 17,39 ± 15,19 ± 16,136 ± 25 and 139 ± 21 in different groups (5,10,20 umol/L siRNA,and controls),respectively (P ＜0.01 ).Conclusion TROP-2 gene might play an important role in adhesion and invasion of human breast cancer cells.siRNA targeted TROP-2 could effectively inhibit adhesion and invasion of human breast cancer cells.