中华外科杂志
中華外科雜誌
중화외과잡지
CHINESE JOURNAL OF SURGERY
2014年
7期
518-522
,共5页
盛伟伟%董明%周建平%柳青峰%李新%董齐
盛偉偉%董明%週建平%柳青峰%李新%董齊
성위위%동명%주건평%류청봉%리신%동제
胰腺肿瘤%肿瘤侵润%肿瘤转移%脑胶质瘤相关癌基因-1
胰腺腫瘤%腫瘤侵潤%腫瘤轉移%腦膠質瘤相關癌基因-1
이선종류%종류침윤%종류전이%뇌효질류상관암기인-1
Pancreatic neoplasms%Neoplasm invasiveness%Neoplasm metastasis%Glioma-associated oncogene-1
目的 探讨脑胶质瘤相关癌基因-1(Gli1)在胰腺癌细胞侵袭和迁移中的作用及可能机制.方法 体外培养胰腺癌细胞株Capan-2,通过RNA干扰技术分别干扰Capan-2细胞中的Gli1、小鼠双微体基因2(MDM2)和p53基因的表达.应用实时定量(qRT)-PCR试验进行干扰效应的验证;通过细胞侵袭和迁移实验观察干扰Capan-2细胞中Gli1、MDM2和p53表达对胰腺癌细胞侵袭和迁移的影响.同时通过Western blot法检测干扰Gli1表达后,金属基质蛋白酶-9(MMP-9)、磷酸化细胞外信号调节蛋白激酶(pERK)及磷酸化蛋白激酶B(pAKT)在Capan-2细胞中表达的变化.采用配对t检验对实验数据进行统计学分析.结果 qRT-PCR试验结果显示,干扰Gli1、MDM2、p53表达后,其mRNA水平较仅加脂质体组和对照组相比分别下调了70.5％和74.5％、61.8％和65.3％、73.8％和78.2％.在p53野生型Capan-2细胞中,干扰Gli1表达后胰腺癌细胞侵袭(94±8)和迁移能力(143±8)较对照组(150 ±7,190±10)均明显减弱(=6.584,P=0.022;t=8.266,P=0.014);干扰MDM2表达亦降低了细胞的侵袭(实验组:85±12,对照组:138 ±6)和迁移能力(实验组:127 ±9,对照组:180±10)(t=5.097,P=0.036;t =4.860,P=0.040).然而,体外干扰p53基因表达后,Capan-2细胞的侵袭(153 ±11)和迁移能力(209±13)较干扰前(106 ±7,164±8)却明显增强(t=4.669,P=0.043;t=4.990,P=0.038).Western blot法检测结果显示,体外干扰Gli1表达后MMP-9蛋白表达水平下调,但pERK和pAKT蛋白水平表达无改变.结论 Gli1可能通过调控MDM2、p53及MMP-9的表达水平促进胰腺癌细胞的侵袭和迁移.
目的 探討腦膠質瘤相關癌基因-1(Gli1)在胰腺癌細胞侵襲和遷移中的作用及可能機製.方法 體外培養胰腺癌細胞株Capan-2,通過RNA榦擾技術分彆榦擾Capan-2細胞中的Gli1、小鼠雙微體基因2(MDM2)和p53基因的錶達.應用實時定量(qRT)-PCR試驗進行榦擾效應的驗證;通過細胞侵襲和遷移實驗觀察榦擾Capan-2細胞中Gli1、MDM2和p53錶達對胰腺癌細胞侵襲和遷移的影響.同時通過Western blot法檢測榦擾Gli1錶達後,金屬基質蛋白酶-9(MMP-9)、燐痠化細胞外信號調節蛋白激酶(pERK)及燐痠化蛋白激酶B(pAKT)在Capan-2細胞中錶達的變化.採用配對t檢驗對實驗數據進行統計學分析.結果 qRT-PCR試驗結果顯示,榦擾Gli1、MDM2、p53錶達後,其mRNA水平較僅加脂質體組和對照組相比分彆下調瞭70.5％和74.5％、61.8％和65.3％、73.8％和78.2％.在p53野生型Capan-2細胞中,榦擾Gli1錶達後胰腺癌細胞侵襲(94±8)和遷移能力(143±8)較對照組(150 ±7,190±10)均明顯減弱(=6.584,P=0.022;t=8.266,P=0.014);榦擾MDM2錶達亦降低瞭細胞的侵襲(實驗組:85±12,對照組:138 ±6)和遷移能力(實驗組:127 ±9,對照組:180±10)(t=5.097,P=0.036;t =4.860,P=0.040).然而,體外榦擾p53基因錶達後,Capan-2細胞的侵襲(153 ±11)和遷移能力(209±13)較榦擾前(106 ±7,164±8)卻明顯增彊(t=4.669,P=0.043;t=4.990,P=0.038).Western blot法檢測結果顯示,體外榦擾Gli1錶達後MMP-9蛋白錶達水平下調,但pERK和pAKT蛋白水平錶達無改變.結論 Gli1可能通過調控MDM2、p53及MMP-9的錶達水平促進胰腺癌細胞的侵襲和遷移.
목적 탐토뇌효질류상관암기인-1(Gli1)재이선암세포침습화천이중적작용급가능궤제.방법 체외배양이선암세포주Capan-2,통과RNA간우기술분별간우Capan-2세포중적Gli1、소서쌍미체기인2(MDM2)화p53기인적표체.응용실시정량(qRT)-PCR시험진행간우효응적험증;통과세포침습화천이실험관찰간우Capan-2세포중Gli1、MDM2화p53표체대이선암세포침습화천이적영향.동시통과Western blot법검측간우Gli1표체후,금속기질단백매-9(MMP-9)、린산화세포외신호조절단백격매(pERK)급린산화단백격매B(pAKT)재Capan-2세포중표체적변화.채용배대t검험대실험수거진행통계학분석.결과 qRT-PCR시험결과현시,간우Gli1、MDM2、p53표체후,기mRNA수평교부가지질체조화대조조상비분별하조료70.5％화74.5％、61.8％화65.3％、73.8％화78.2％.재p53야생형Capan-2세포중,간우Gli1표체후이선암세포침습(94±8)화천이능력(143±8)교대조조(150 ±7,190±10)균명현감약(=6.584,P=0.022;t=8.266,P=0.014);간우MDM2표체역강저료세포적침습(실험조:85±12,대조조:138 ±6)화천이능력(실험조:127 ±9,대조조:180±10)(t=5.097,P=0.036;t =4.860,P=0.040).연이,체외간우p53기인표체후,Capan-2세포적침습(153 ±11)화천이능력(209±13)교간우전(106 ±7,164±8)각명현증강(t=4.669,P=0.043;t=4.990,P=0.038).Western blot법검측결과현시,체외간우Gli1표체후MMP-9단백표체수평하조,단pERK화pAKT단백수평표체무개변.결론 Gli1가능통과조공MDM2、p53급MMP-9적표체수평촉진이선암세포적침습화천이.
Objective To study the role and possible mechanism of glioma-associated oncogene-1 (Gli1) in regulating the cell invasion and migration of pancreatic cancer cells.Methods Quantitative realtime (qRT)-PCR was used to detect the effect of siRNA interference on Gli1,murine double minute 2 (MDM2) and p53 genes.Cell invasion and migration assays were used to observe the effect of Gli1,MDM2 and p53 silence on cell invasion and migration in p53 wild-type Capan-2 pancreatic cancer cells,respectively.Meanwhile,immunoblotting(IB) was used to detect the protein level of matrix metalloproteinase (MMP)-9,phospho-excelluar signal-regulated kinase (pERK) and phosphorylation protein kinase B (pAKT) in Gli1-silencing Capan-2 cells.The data were analyzed by paired t-test.Results qRT-PCR showed that the expression of Gli1,MDM2 and p53 is down-regulated 70.5％ and 74.5％,61.8％ and 65.3％,and 73.8％ and 78.2％ after siRNA interference,compared with the mock and siRNA control groups,respectively.Cell invasion(94 ± 8)and migration(143 ± 8) in p53 wild-type Capan-2 cells transfected with Gli1siRNA were significantly decreased,compared with the siRNA control group (150 ± 7,190 ± 10) (t =6.584,P =0.022 ;t =8.266,P =0.014),while MDM2 silence inhibited cell invasion (experiment group:85 ± 12,control group:138 ± 6)and migration(experiment group:127 ± 9,control group:180 ± 10) in the same cells,respectively(t =5.097,P =0.036;t =4.860,P =0.040).However,cell invasion(experiment group:153 ± 11,control group:106 ± 7) and migration (experiment group:209 ± 13,control group:164 ± 8) in p53-silencing Capan-2 cells were significantly enhanced (t =4.669,P =0.043 ; t =4.990,P =0.038).IB showed that Gli1 silence down-regulated MMP-9 but not pERK and pAKT protein expression.Conclusion Gli1 might contribute to the cell invasion and migration in pancreatic cancer via the regulation of MDM2,p53 and MMP-9 expression.
