中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2010年
8期
1119-1121
,共3页
肺肿瘤%侵袭%基质金属蛋白酶
肺腫瘤%侵襲%基質金屬蛋白酶
폐종류%침습%기질금속단백매
Lung carcinoma%Invasion%Matrix metalloproteinase
目的 探讨促肝细胞再生磷酸酶-1(PRL-1)基因对肺癌细胞侵袭的影响及其机制.方法 应用PRL-1基因小干扰RNA(siRNA)转染处理肺癌细胞株A549后,分别采用实时定量逆转录-聚合酶链反应(RT-PCR)和Western blot检测PRL-1基因和基质金属蛋白酶(MMP)-2、MMP-7和MMP-9 mRNA和蛋白水平,分别采用软琼脂集落培养实验和Boyden小室模型实验检测癌细胞的锚着不依赖性增殖和侵袭能力.结果 siRNA转染组PRL-1 mRNA和蛋白水平明显下调,且呈浓度和时间依赖性(P＜0.01).软琼脂集落形成实验显示,3.125、6.25和12.5 nmol/L siRNA组集落形成数分别为17.8±1.6、13.6±1.5、8.8±1.4,而对照组为22.6±1.8(P＜0.05);Boyden小室模型实验显示,3.125、6.25和12.5 nmol/L siRNA组穿过滤膜的细胞分别为33.6±2.1、19.5±1.9、8.1±1.8,而对照组为49.4±2.3(P＜0.05).同时发现,转染组细胞MMP-2、MMP-7、MMP-9基因mRNA和蛋白水平明显下调.结论 PRL-1 siRNA转染可抑制肺癌细胞侵袭,其机制可能与下调MMP-2、MMP-7、MMP-9表达有关.
目的 探討促肝細胞再生燐痠酶-1(PRL-1)基因對肺癌細胞侵襲的影響及其機製.方法 應用PRL-1基因小榦擾RNA(siRNA)轉染處理肺癌細胞株A549後,分彆採用實時定量逆轉錄-聚閤酶鏈反應(RT-PCR)和Western blot檢測PRL-1基因和基質金屬蛋白酶(MMP)-2、MMP-7和MMP-9 mRNA和蛋白水平,分彆採用軟瓊脂集落培養實驗和Boyden小室模型實驗檢測癌細胞的錨著不依賴性增殖和侵襲能力.結果 siRNA轉染組PRL-1 mRNA和蛋白水平明顯下調,且呈濃度和時間依賴性(P＜0.01).軟瓊脂集落形成實驗顯示,3.125、6.25和12.5 nmol/L siRNA組集落形成數分彆為17.8±1.6、13.6±1.5、8.8±1.4,而對照組為22.6±1.8(P＜0.05);Boyden小室模型實驗顯示,3.125、6.25和12.5 nmol/L siRNA組穿過濾膜的細胞分彆為33.6±2.1、19.5±1.9、8.1±1.8,而對照組為49.4±2.3(P＜0.05).同時髮現,轉染組細胞MMP-2、MMP-7、MMP-9基因mRNA和蛋白水平明顯下調.結論 PRL-1 siRNA轉染可抑製肺癌細胞侵襲,其機製可能與下調MMP-2、MMP-7、MMP-9錶達有關.
목적 탐토촉간세포재생린산매-1(PRL-1)기인대폐암세포침습적영향급기궤제.방법 응용PRL-1기인소간우RNA(siRNA)전염처리폐암세포주A549후,분별채용실시정량역전록-취합매련반응(RT-PCR)화Western blot검측PRL-1기인화기질금속단백매(MMP)-2、MMP-7화MMP-9 mRNA화단백수평,분별채용연경지집락배양실험화Boyden소실모형실험검측암세포적묘착불의뢰성증식화침습능력.결과 siRNA전염조PRL-1 mRNA화단백수평명현하조,차정농도화시간의뢰성(P＜0.01).연경지집락형성실험현시,3.125、6.25화12.5 nmol/L siRNA조집락형성수분별위17.8±1.6、13.6±1.5、8.8±1.4,이대조조위22.6±1.8(P＜0.05);Boyden소실모형실험현시,3.125、6.25화12.5 nmol/L siRNA조천과려막적세포분별위33.6±2.1、19.5±1.9、8.1±1.8,이대조조위49.4±2.3(P＜0.05).동시발현,전염조세포MMP-2、MMP-7、MMP-9기인mRNA화단백수평명현하조.결론 PRL-1 siRNA전염가억제폐암세포침습,기궤제가능여하조MMP-2、MMP-7、MMP-9표체유관.
Objective To study the effects of phosphatase of regenerating liver cell-1 (PRL-1) small interfering RNA (siRNA) on invasion of lung cancer cells and the mechanism. Methods After lung cancer cell line A549 cells were transfected by PRL-1 siRNA, the mRNA and protein expression levels of PRL-1 and matrix metalloproteinase-2 and-9 were detected by real time reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting, respectively. The anchorage-independent growth was examined using colon formation in soft agar, and invasion ability was evaluated by boyden chamber model. Results The anchorage-independent growth showed that the colonies were 17.8 ± 1.6, 13.6 ± 1.5, 8.8 ± 1.4, and 23.6 ±1.8 in different groups (3.125, 6.25, 12.5 nmol/L siRNA and control), respectively. The boyden chamber results revealed that the number of invasion cells was 33.6 ± 3.1, 19.5 ±1.9, 8.1 ±1.8 and 49.4 ±3.3 in different groups (3.125, 6.25, 13.5 nmol/L siRNA, and control) , respectively. The mRNA and protein expression levels of MMP-2,-7 and-9 in transfection group with PRL-1 siRNA were down-regulated as compared with control group in lung cancer cells. Conclusion PRL-1 siRNA can inhibit invasion through down-regulating MMPs of lung cancer cells.
