中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2014年
6期
1228-1230
,共3页
周鹏飞%王大文%南存金%吴铁林%杨森%陈映鹤
週鵬飛%王大文%南存金%吳鐵林%楊森%陳映鶴
주붕비%왕대문%남존금%오철림%양삼%진영학
前列腺癌%肝细胞再生磷酸酶-3%RNA干扰
前列腺癌%肝細胞再生燐痠酶-3%RNA榦擾
전렬선암%간세포재생린산매-3%RNA간우
Prostate cancer%Phosphatase of regenerating liver cell 3%RNA interference
目的 观察小干扰RNA(siRNA)沉默促肝细胞再生磷酸酶-3(PRL-3)基因对人雄激素依赖性前列腺癌(LNCaP)细胞增殖及凋亡的影响.方法 应用靶向PRL-3基因小干扰RNA转染处理前列腺癌LNCaP细胞后,采用逆转录-聚合酶链反应(RT-PCR)和Western blot检测PRL-3 mRNA和蛋白水平,采用细胞计数试剂盒(CCK-8)检测细胞体外增殖能力,采用流式细胞仪检测细胞凋亡水平.结果 与对照组比较,siRNA转染组PRL-3 mRNA和蛋白水平明显下调,且呈浓度依赖性(P＜0.05).体外实验显示,低浓度PRL-3 siRNA对LNCaP细胞体外增殖能力没有影响,而中、高浓度PRL-3 siRNA转染48 h后明显地抑制LNCaP细胞的体外增殖能力(P＜0.05),低、中、高浓度siRNA沉默PRL-3基因后体外LNCaP细胞的凋亡率分别为(28.1±3.8)％、(25.2±2.5)％、(27.1±0.7)％(P＜0.05).结论 下调PRL-3基因的表达可抑制LNCaP细胞的增殖并促进LNCaP细胞的凋亡,PRL-3基因在前列腺癌LNCaP细胞体外增殖及凋亡中起重要作用.
目的 觀察小榦擾RNA(siRNA)沉默促肝細胞再生燐痠酶-3(PRL-3)基因對人雄激素依賴性前列腺癌(LNCaP)細胞增殖及凋亡的影響.方法 應用靶嚮PRL-3基因小榦擾RNA轉染處理前列腺癌LNCaP細胞後,採用逆轉錄-聚閤酶鏈反應(RT-PCR)和Western blot檢測PRL-3 mRNA和蛋白水平,採用細胞計數試劑盒(CCK-8)檢測細胞體外增殖能力,採用流式細胞儀檢測細胞凋亡水平.結果 與對照組比較,siRNA轉染組PRL-3 mRNA和蛋白水平明顯下調,且呈濃度依賴性(P＜0.05).體外實驗顯示,低濃度PRL-3 siRNA對LNCaP細胞體外增殖能力沒有影響,而中、高濃度PRL-3 siRNA轉染48 h後明顯地抑製LNCaP細胞的體外增殖能力(P＜0.05),低、中、高濃度siRNA沉默PRL-3基因後體外LNCaP細胞的凋亡率分彆為(28.1±3.8)％、(25.2±2.5)％、(27.1±0.7)％(P＜0.05).結論 下調PRL-3基因的錶達可抑製LNCaP細胞的增殖併促進LNCaP細胞的凋亡,PRL-3基因在前列腺癌LNCaP細胞體外增殖及凋亡中起重要作用.
목적 관찰소간우RNA(siRNA)침묵촉간세포재생린산매-3(PRL-3)기인대인웅격소의뢰성전렬선암(LNCaP)세포증식급조망적영향.방법 응용파향PRL-3기인소간우RNA전염처리전렬선암LNCaP세포후,채용역전록-취합매련반응(RT-PCR)화Western blot검측PRL-3 mRNA화단백수평,채용세포계수시제합(CCK-8)검측세포체외증식능력,채용류식세포의검측세포조망수평.결과 여대조조비교,siRNA전염조PRL-3 mRNA화단백수평명현하조,차정농도의뢰성(P＜0.05).체외실험현시,저농도PRL-3 siRNA대LNCaP세포체외증식능력몰유영향,이중、고농도PRL-3 siRNA전염48 h후명현지억제LNCaP세포적체외증식능력(P＜0.05),저、중、고농도siRNA침묵PRL-3기인후체외LNCaP세포적조망솔분별위(28.1±3.8)％、(25.2±2.5)％、(27.1±0.7)％(P＜0.05).결론 하조PRL-3기인적표체가억제LNCaP세포적증식병촉진LNCaP세포적조망,PRL-3기인재전렬선암LNCaP세포체외증식급조망중기중요작용.
Objective To study the effects of small interfering RNA (siRNA) silencing phosphatase of regenerating liver cell 3 (PRL-3) on proliferation and apoptosis of human prostate cancer LNCaP cells.Methods After prostate cancer LNCaP cells were transfected by PRL-3 siRNA,the mRNA and protein of PRL-3 were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting,respectively.The growth of LNCaP cells was exmined by cell counting kit-8 (CCK-8).Flow cytometry was used to detect apoptosis.Results The siRNA could downregulate the mRNA and protein expression level of PRL-3 in a dose-dependent manner (P ＜ 0.05).Low doses of PRL-3 siRNA did not affect the proliferation of LNCaP cells,but 100 nmol/L PRL-3 siRNA could effectively inhibit the proliferation of LNCaP cells after 48 h in vitro (P ＜ 0.05).Apoptosis rate of LNCaP cells with low,middle and high PRL3 siRNA interventions in vitro was (28.1 ± 3.8) ％,(25.2 ± 2.5) ％,and (27.1 ± 0.7) ％ respectively (P ＜ 0.05).Conclusion Downregulating PRL-3 could inhibit proliferation of LNCaP cells and promote apoptosis of LNCaP cells.PRL-3 gene might play an important role in proliferation and apoptosis of LNCaP cells.
