中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
12期
2500-2502
,共3页
崔飞伦%满昌峰%周永静%范钰
崔飛倫%滿昌峰%週永靜%範鈺
최비륜%만창봉%주영정%범옥
前列腺肿瘤%人滋养层细胞表面抗原-2基因%侵袭%尿激酶纤溶酶原激活物
前列腺腫瘤%人滋養層細胞錶麵抗原-2基因%侵襲%尿激酶纖溶酶原激活物
전렬선종류%인자양층세포표면항원-2기인%침습%뇨격매섬용매원격활물
Prostate carcinoma%Trophoblast cell-surface antigens 2 gene%Invasion%Urokinase-type plasminogen activator
目的 观察人滋养层细胞表面抗原-2(Trop-2)基因对人前列腺细胞侵袭的影响,并探讨其分子机制.方法 采用TROP-2基因小于扰RNA(siRNA)转染处理人前列腺PC-3细胞后,分别采用实时荧光定量聚合酶链反应(FQ-PCR)和Western blot检测TROP-2基因mRNA和蛋白水平;采用Transwell小室法试验检测癌细胞侵袭能力;采用Western blot法检测癌细胞尿激酶纤溶酶原激活物(uPA)蛋白变化.结果 siRNA转染组细胞TROP-2基因mRNA和蛋白水平明显下调,且呈浓度依赖性(P＜0.01).siRNA转染组穿过滤膜的细胞数明显下降,且呈浓度依赖性(P＜0.01).Transwell小室试验结果显示,Con-A、Con-B、siRNA(5nmol/L)、siRNA(10 nmol/L)、siRNA(20 nmol/L)组穿膜细胞数分别为(128.16 ±3.89)、(127.58 ±3.56)、(102.56 ±3.28)、(76.38 ±3.25)和(39.89±3.18)个.Western blot结果显示,TROP-2 siRNA转染组uPA蛋白表达明显下调,且与浓度相关.结论 TROP-2基因在人前列腺癌细胞侵袭中发挥重要的作用,采用siRNA转染可抑制前列腺细胞侵袭,下调uPA表达,是其重要机制之一.
目的 觀察人滋養層細胞錶麵抗原-2(Trop-2)基因對人前列腺細胞侵襲的影響,併探討其分子機製.方法 採用TROP-2基因小于擾RNA(siRNA)轉染處理人前列腺PC-3細胞後,分彆採用實時熒光定量聚閤酶鏈反應(FQ-PCR)和Western blot檢測TROP-2基因mRNA和蛋白水平;採用Transwell小室法試驗檢測癌細胞侵襲能力;採用Western blot法檢測癌細胞尿激酶纖溶酶原激活物(uPA)蛋白變化.結果 siRNA轉染組細胞TROP-2基因mRNA和蛋白水平明顯下調,且呈濃度依賴性(P＜0.01).siRNA轉染組穿過濾膜的細胞數明顯下降,且呈濃度依賴性(P＜0.01).Transwell小室試驗結果顯示,Con-A、Con-B、siRNA(5nmol/L)、siRNA(10 nmol/L)、siRNA(20 nmol/L)組穿膜細胞數分彆為(128.16 ±3.89)、(127.58 ±3.56)、(102.56 ±3.28)、(76.38 ±3.25)和(39.89±3.18)箇.Western blot結果顯示,TROP-2 siRNA轉染組uPA蛋白錶達明顯下調,且與濃度相關.結論 TROP-2基因在人前列腺癌細胞侵襲中髮揮重要的作用,採用siRNA轉染可抑製前列腺細胞侵襲,下調uPA錶達,是其重要機製之一.
목적 관찰인자양층세포표면항원-2(Trop-2)기인대인전렬선세포침습적영향,병탐토기분자궤제.방법 채용TROP-2기인소우우RNA(siRNA)전염처리인전렬선PC-3세포후,분별채용실시형광정량취합매련반응(FQ-PCR)화Western blot검측TROP-2기인mRNA화단백수평;채용Transwell소실법시험검측암세포침습능력;채용Western blot법검측암세포뇨격매섬용매원격활물(uPA)단백변화.결과 siRNA전염조세포TROP-2기인mRNA화단백수평명현하조,차정농도의뢰성(P＜0.01).siRNA전염조천과려막적세포수명현하강,차정농도의뢰성(P＜0.01).Transwell소실시험결과현시,Con-A、Con-B、siRNA(5nmol/L)、siRNA(10 nmol/L)、siRNA(20 nmol/L)조천막세포수분별위(128.16 ±3.89)、(127.58 ±3.56)、(102.56 ±3.28)、(76.38 ±3.25)화(39.89±3.18)개.Western blot결과현시,TROP-2 siRNA전염조uPA단백표체명현하조,차여농도상관.결론 TROP-2기인재인전렬선암세포침습중발휘중요적작용,채용siRNA전염가억제전렬선세포침습,하조uPA표체,시기중요궤제지일.
Objective To explore the effects and mechanism of trophoblast cell-surface antigens 2 (TROP-2) on invasion of human prostate cancer cells.Methods After human prostate cancer PC-3 cell line was transfected with TROP-2 small interfering RNA (siRNA),the mRNA and protein levels of TROP-2 were determined by using real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) and Western blotting respectively.The invasion ability was evaluated by Transwell.The urokinase-type plasminogen activator (uPA) protein of cancer cells was examined by Western blotting.Results The mRNA and protein expression levels of TROP-2 were greatly inhibited in PC-3 cancer cells transfected with TROP-2 siRNA.The results of the Transwell assay showed that the number of cells penetrating the membrane in Con-A,Con-B,and siRNA groups (5,10 and 20 nmoL/L) was 128.16 ± 3.89,127.58 ± 3.56,102.56 ± 3.28,76.38 ± 3.25,and 39.89 ±3.18,respectively (P ＜0.05).The results from Western blotting assay showed that the uPA protein in siRNA groups was reduced significantly.Conclusion TROP-2 siRNA inhibit the invasion of prostate cancer cells through down-regulation of uPA.
