中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2009年
10期
732-737
,共6页
caveolin-1%胰腺肿瘤%受体%表皮生长因子
caveolin-1%胰腺腫瘤%受體%錶皮生長因子
caveolin-1%이선종류%수체%표피생장인자
Caveolin-1%Pancreatic neoplasms%Receptor,epidermal growth factor
目的 体外研究caveolin-1对胰腺癌细胞生物学行为及相关信号通路的影响.方法 caveolin-1基因脂质体法稳定转染胰腺癌细胞株panc1,筛选出稳定高表达caveolin-1的克隆,并用实时荧光定量PCR和免疫印迹法鉴定.应用四甲基偶氮唑蓝(MTT)法检测细胞的生长能力,软琼脂克隆生长实验检测细胞锚定非依赖性生长能力,流式细胞仪检测细胞周期和凋亡情况,细胞侵袭实验检测细胞侵袭能力,免疫印迹法检测表皮生长因子受体(EGFR)、c-Raf、Mek、Erk、p38和SAPK/JNK的表达.结果 转染panc1细胞后筛选到3株稳定高表达caveolin-1的克隆.MTT法检测结果 显示,转染后的细胞生长速度明显慢于对照组,软琼脂中克隆生长能力较对照组也明显下降.流式细胞术检测结果 显示,高表达caveolin-1可使细胞阻滞在G0/G1期,并且促进细胞的凋亡.细胞侵袭实验显示,转染caveolin-1后,细胞的侵袭能力明显下降,此外,caveolin-1过表达降低了EGFR、c-Raf、Mek和Erk的磷酸化水平.结论 caveolin-1基因过表达抑制了胰腺癌细胞panc1的生长和侵袭能力,对EGFR-c-Raf-Mek-Erk信号级联的抑制与其抑制胰腺癌细胞恶性表型的作用相关.
目的 體外研究caveolin-1對胰腺癌細胞生物學行為及相關信號通路的影響.方法 caveolin-1基因脂質體法穩定轉染胰腺癌細胞株panc1,篩選齣穩定高錶達caveolin-1的剋隆,併用實時熒光定量PCR和免疫印跡法鑒定.應用四甲基偶氮唑藍(MTT)法檢測細胞的生長能力,軟瓊脂剋隆生長實驗檢測細胞錨定非依賴性生長能力,流式細胞儀檢測細胞週期和凋亡情況,細胞侵襲實驗檢測細胞侵襲能力,免疫印跡法檢測錶皮生長因子受體(EGFR)、c-Raf、Mek、Erk、p38和SAPK/JNK的錶達.結果 轉染panc1細胞後篩選到3株穩定高錶達caveolin-1的剋隆.MTT法檢測結果 顯示,轉染後的細胞生長速度明顯慢于對照組,軟瓊脂中剋隆生長能力較對照組也明顯下降.流式細胞術檢測結果 顯示,高錶達caveolin-1可使細胞阻滯在G0/G1期,併且促進細胞的凋亡.細胞侵襲實驗顯示,轉染caveolin-1後,細胞的侵襲能力明顯下降,此外,caveolin-1過錶達降低瞭EGFR、c-Raf、Mek和Erk的燐痠化水平.結論 caveolin-1基因過錶達抑製瞭胰腺癌細胞panc1的生長和侵襲能力,對EGFR-c-Raf-Mek-Erk信號級聯的抑製與其抑製胰腺癌細胞噁性錶型的作用相關.
목적 체외연구caveolin-1대이선암세포생물학행위급상관신호통로적영향.방법 caveolin-1기인지질체법은정전염이선암세포주panc1,사선출은정고표체caveolin-1적극륭,병용실시형광정량PCR화면역인적법감정.응용사갑기우담서람(MTT)법검측세포적생장능력,연경지극륭생장실험검측세포묘정비의뢰성생장능력,류식세포의검측세포주기화조망정황,세포침습실험검측세포침습능력,면역인적법검측표피생장인자수체(EGFR)、c-Raf、Mek、Erk、p38화SAPK/JNK적표체.결과 전염panc1세포후사선도3주은정고표체caveolin-1적극륭.MTT법검측결과 현시,전염후적세포생장속도명현만우대조조,연경지중극륭생장능력교대조조야명현하강.류식세포술검측결과 현시,고표체caveolin-1가사세포조체재G0/G1기,병차촉진세포적조망.세포침습실험현시,전염caveolin-1후,세포적침습능력명현하강,차외,caveolin-1과표체강저료EGFR、c-Raf、Mek화Erk적린산화수평.결론 caveolin-1기인과표체억제료이선암세포panc1적생장화침습능력,대EGFR-c-Raf-Mek-Erk신호급련적억제여기억제이선암세포악성표형적작용상관.
Objective To investigate the effects of caveolin-1 on the biologic behavior of pancreatic carcinoma cell line panel cells in vitro.Methods Eukaryotic expression vectors containing human caveolin-1 gene was stably transfected into panc1 cells with Lipofectamine2000.The clones stably overexpressing caveolin-1 were identified by real-time PCR and Western bloting.The cell growth activity was examined by MTT assay.Anchorage-independent growth was detected by colony formation assay in soft agar.Flow cytometry was used to analyze the cell cycle and apoptosis.Cell invasion assay was used for evaluating cell invasion capacity.The relative phosphorylation level of EGFR,c-Raf,Mek,Erk,p38 and SAPK/JNK were detected by Western blotting.Results Three transfected cell clones overexpressing caveolin-1 were obtained.Comparing with the panel cells,the transfeeted cells exhibited a slower growth rate and formed fewer colonies in soft agar.The results of flow cytometry showed that over-expression of caveolin-1 resulted in the cell cycle arrest at G0/G1 phase and increased the apoptotic cell fraction.Cell invasion assay showed that overexpression of caveolin-1 significantly inhibited the panel cell invasion.Western blotting results showed that overexpression of caveolin-1 reduced the phosphorylation of EGFR,c-Raf,Mek and Erk while did not affect the activity of p38 and SAPK/JNK.Conclusion Over-expression of caveolin-1 inhibits the growth and invasion of pancreatic carcinoma cells in vitro.These phenotypes may be correlated with the inhibition of EGFR-c-Raf-Mek-Erk signaling pathway.
