中华肝胆外科杂志
中華肝膽外科雜誌
중화간담외과잡지
CHINESE JOURNAL OF HEPATOBILIARY SURGERY
2011年
1期
31-35
,共5页
张克君%余壮明%王正文%孙传东%李德春%张炳远%卢云%赵伟
張剋君%餘壯明%王正文%孫傳東%李德春%張炳遠%盧雲%趙偉
장극군%여장명%왕정문%손전동%리덕춘%장병원%로운%조위
胰腺肿瘤%转录因子%钙黏着糖蛋白类%转染%肿瘤转移
胰腺腫瘤%轉錄因子%鈣黏著糖蛋白類%轉染%腫瘤轉移
이선종류%전록인자%개점착당단백류%전염%종류전이
Pancreatic carcinoma%Slug%Cadherins%Transfection%Metastasis
目的 探讨Slug和E-CADHERIN(E-cadherin)表达与胰腺癌转移的关系以及干扰Slug转录因子对胰腺癌转移的影响.方法 采用免疫组化和RT-PCR技术分别检测36例胰腺癌组织中Slug、E-cadherin和mRNA表达.将肝脏高转移潜能的胰腺癌细胞株SW1990H4,分为3组:对照组、空载体质粒(shRNA-1)和转染干扰S1ug质粒(Slug-shRNA-1),观察Slug对E-cadherin mRNA表达的逆转作用,及对SW1990H4体外侵袭、运动的抑制作用.结果 胰腺癌组织中Slug高表达,而E-cadherin低表达.转移组胰腺癌组织中Slug蛋白和mRNA表达明显高于非转移组,差异分别有统计学意义(P＜0.05),而E-cadherin在转移组胰腺癌中无表达.PCR产物凝胶电泳结果显示,Slug mRNA在空白对照组SW1990H4中表达为0.985±0.016,E-cadherin mRNA表达为0.120±0.001.shRNA-1时转染48 h时,Slug mRNA的表达水平为0.973±0.014,E-cadherin mRNA表达水平分别0.160±0.001,与对照组比差异分别无统计学意义(P＞0.05).转染Slug-shRNA-1组瞬时转染48 h时,Slug mRNA的表达水平为0.554±0.011,E-cadherin mRNA表达水平为0.36±0.002,与对照组和shRNA-1组比差异分别有统计学意义(P＜0.05).SW1990H4稳定转染后,shRNA-1和Slug-shRNA-1组Slug mRNA表达水平分别为0.968±0.015,0.206±0.017,两组比较,差异有统计学意义(P＜0.05),而E-cadherin mRNA表达水平分别为0.18±0.002,0.727±0.006,两组比较,差异有统计学意义(P＜0.05).体外细胞运动实验表明,对照组、转染shRNA-1和SlugshRNA-1组跨膜细胞数393±28、352±24、96±13,差异有统计学意义(P＜0.01).重组细胞基底膜(Matrige1)侵袭实验显示,3组穿透基底膜细胞数分别为223±69、202±64、65±19,差异有统计学意义(P＜0.05).结论 Slug高表达,E-cadherin低表达与胰腺癌的转移相关.胰腺癌细胞中存在Slug mRNA与E-cadherin mRNA表达的逆转关系,抑制Slug mRNA的表达对胰腺癌细胞的侵袭和运动有抑制作用,可能为胰腺癌转移的基因治疗提供新的靶点.
目的 探討Slug和E-CADHERIN(E-cadherin)錶達與胰腺癌轉移的關繫以及榦擾Slug轉錄因子對胰腺癌轉移的影響.方法 採用免疫組化和RT-PCR技術分彆檢測36例胰腺癌組織中Slug、E-cadherin和mRNA錶達.將肝髒高轉移潛能的胰腺癌細胞株SW1990H4,分為3組:對照組、空載體質粒(shRNA-1)和轉染榦擾S1ug質粒(Slug-shRNA-1),觀察Slug對E-cadherin mRNA錶達的逆轉作用,及對SW1990H4體外侵襲、運動的抑製作用.結果 胰腺癌組織中Slug高錶達,而E-cadherin低錶達.轉移組胰腺癌組織中Slug蛋白和mRNA錶達明顯高于非轉移組,差異分彆有統計學意義(P＜0.05),而E-cadherin在轉移組胰腺癌中無錶達.PCR產物凝膠電泳結果顯示,Slug mRNA在空白對照組SW1990H4中錶達為0.985±0.016,E-cadherin mRNA錶達為0.120±0.001.shRNA-1時轉染48 h時,Slug mRNA的錶達水平為0.973±0.014,E-cadherin mRNA錶達水平分彆0.160±0.001,與對照組比差異分彆無統計學意義(P＞0.05).轉染Slug-shRNA-1組瞬時轉染48 h時,Slug mRNA的錶達水平為0.554±0.011,E-cadherin mRNA錶達水平為0.36±0.002,與對照組和shRNA-1組比差異分彆有統計學意義(P＜0.05).SW1990H4穩定轉染後,shRNA-1和Slug-shRNA-1組Slug mRNA錶達水平分彆為0.968±0.015,0.206±0.017,兩組比較,差異有統計學意義(P＜0.05),而E-cadherin mRNA錶達水平分彆為0.18±0.002,0.727±0.006,兩組比較,差異有統計學意義(P＜0.05).體外細胞運動實驗錶明,對照組、轉染shRNA-1和SlugshRNA-1組跨膜細胞數393±28、352±24、96±13,差異有統計學意義(P＜0.01).重組細胞基底膜(Matrige1)侵襲實驗顯示,3組穿透基底膜細胞數分彆為223±69、202±64、65±19,差異有統計學意義(P＜0.05).結論 Slug高錶達,E-cadherin低錶達與胰腺癌的轉移相關.胰腺癌細胞中存在Slug mRNA與E-cadherin mRNA錶達的逆轉關繫,抑製Slug mRNA的錶達對胰腺癌細胞的侵襲和運動有抑製作用,可能為胰腺癌轉移的基因治療提供新的靶點.
목적 탐토Slug화E-CADHERIN(E-cadherin)표체여이선암전이적관계이급간우Slug전록인자대이선암전이적영향.방법 채용면역조화화RT-PCR기술분별검측36례이선암조직중Slug、E-cadherin화mRNA표체.장간장고전이잠능적이선암세포주SW1990H4,분위3조:대조조、공재체질립(shRNA-1)화전염간우S1ug질립(Slug-shRNA-1),관찰Slug대E-cadherin mRNA표체적역전작용,급대SW1990H4체외침습、운동적억제작용.결과 이선암조직중Slug고표체,이E-cadherin저표체.전이조이선암조직중Slug단백화mRNA표체명현고우비전이조,차이분별유통계학의의(P＜0.05),이E-cadherin재전이조이선암중무표체.PCR산물응효전영결과현시,Slug mRNA재공백대조조SW1990H4중표체위0.985±0.016,E-cadherin mRNA표체위0.120±0.001.shRNA-1시전염48 h시,Slug mRNA적표체수평위0.973±0.014,E-cadherin mRNA표체수평분별0.160±0.001,여대조조비차이분별무통계학의의(P＞0.05).전염Slug-shRNA-1조순시전염48 h시,Slug mRNA적표체수평위0.554±0.011,E-cadherin mRNA표체수평위0.36±0.002,여대조조화shRNA-1조비차이분별유통계학의의(P＜0.05).SW1990H4은정전염후,shRNA-1화Slug-shRNA-1조Slug mRNA표체수평분별위0.968±0.015,0.206±0.017,량조비교,차이유통계학의의(P＜0.05),이E-cadherin mRNA표체수평분별위0.18±0.002,0.727±0.006,량조비교,차이유통계학의의(P＜0.05).체외세포운동실험표명,대조조、전염shRNA-1화SlugshRNA-1조과막세포수393±28、352±24、96±13,차이유통계학의의(P＜0.01).중조세포기저막(Matrige1)침습실험현시,3조천투기저막세포수분별위223±69、202±64、65±19,차이유통계학의의(P＜0.05).결론 Slug고표체,E-cadherin저표체여이선암적전이상관.이선암세포중존재Slug mRNA여E-cadherin mRNA표체적역전관계,억제Slug mRNA적표체대이선암세포적침습화운동유억제작용,가능위이선암전이적기인치료제공신적파점.
Objective To investigate expression of slug and E-cadherin in pancreatic cancer tissues and determine the inhibitory effects of anti-Slug, an anti-sense plasmid, on the invasion of pancreatic cancer cell lines in vitro. Methods Slug and E-cadherin protein and mRNA was analyzed by IHP and RT-PCR in 36 cases of pancreatic cancer. Then anti-Slug plasmid was transfected into herin and Slug expression. The inhibitory effects of anti-sense Slug were also detected by Transwell motility assay and Matrigel invasion assay. Results The expression of Slug and mRNA in metastatic pancreatic cancer tissue was higher than that in non-metastatic tissue. E-cadherin and mRNA was lower in metastasis tissues(P＜0.05). The inverse relationships were further observed by transient transfection of anti-Slug into SW1990H4 cells. The downregulated expression of Slug and re-expression of E-cadherin were found. The Slug mRNA levels were 0.985±0.016,0.973±0.014, 0. 554±0. 011 after 0, 48 h of transfection of anti-sense Slug, and that of E-cadherin were 0.120±0.001, 0.360±0.002, 0. 727±0. 006, respectively. The diference was significant between different time points (P＜0.05). The Slug mRNA levels were 0. 206±0.017, 0.968±0.015, and that of E-cadherin were 0. 18±0.002,0.727±0.006 after stable transfection of anti-sense Slug, and control plasmid, respectively. The diference was significant (P＜0.05). The motility activity(393±28, 352±24, 96 ±13 )and the invasion activity (223 ± 69, 202 ± 64, 65 ±19) of1 antisense Slug transfectant cells were significantly decreased as compared with those of control cells (P＜0.05). Conclusions Higher expression of slug and lower expression of E-cadherin is related to the invasion and metastasis in pancreatic cancer. A reverse corelation of E-cadherin and Slug expression exists in pancreatic cancer. Slug is possibly a potential target for cancer gene therapy blocking invasion and metastasis in human pancreatic cancer.
