中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2012年
11期
2175-2177
,共3页
张靖%许志朋%朱金水%周洲%陈维雄%陈尼维
張靖%許誌朋%硃金水%週洲%陳維雄%陳尼維
장정%허지붕%주금수%주주%진유웅%진니유
胃癌%Yes相关蛋白%小发夹RNA%增殖%转移
胃癌%Yes相關蛋白%小髮夾RNA%增殖%轉移
위암%Yes상관단백%소발협RNA%증식%전이
Gastric carcinoma%Yes-associated protein%Small hairpin RNA%Proliferation%Metastasis
目的 检测Yes相关蛋白(YAP)在人胃腺癌、胃腺瘤及正常胃黏膜组织中的表达,探讨YAP对胃癌细胞增殖和转移的作用及其分子机制.方法 免疫组织化学法和实时定量聚合酶链反应(Real-time PCR)检测YAP在78例人胃腺癌、39例人胃腺瘤及46例正常人胃黏膜组织的表达.利用建立慢病毒小发夹RNA(shRNA)靶向YAP基因的胃癌SGC-7901细胞株,Western blot法检测增殖细胞核抗原(PCNA)和基质金属蛋白酶-2(MMP-2)的蛋白表达水平.噻唑蓝(MTT)比色法和Transwell试验检测YAP下调对胃癌细胞增殖和转移的影响.结果 YAP在69.23％的胃腺癌组织中高表达,与正常胃黏膜(0.33 ±0.15)和胃腺瘤(0.42 ±0.20)比较,YAP mRNA在胃腺癌(1.74±0.46)的表达水平明显增高(P＜0.001).体外实验显示,YAP下调减少PCNA和MMP-2蛋白的表达,并抑制胃癌细胞增殖活性和转移能力.结论 YAP在人胃腺癌组织中高表达,并且YAP下调抑制胃癌细胞的增殖和转移.
目的 檢測Yes相關蛋白(YAP)在人胃腺癌、胃腺瘤及正常胃黏膜組織中的錶達,探討YAP對胃癌細胞增殖和轉移的作用及其分子機製.方法 免疫組織化學法和實時定量聚閤酶鏈反應(Real-time PCR)檢測YAP在78例人胃腺癌、39例人胃腺瘤及46例正常人胃黏膜組織的錶達.利用建立慢病毒小髮夾RNA(shRNA)靶嚮YAP基因的胃癌SGC-7901細胞株,Western blot法檢測增殖細胞覈抗原(PCNA)和基質金屬蛋白酶-2(MMP-2)的蛋白錶達水平.噻唑藍(MTT)比色法和Transwell試驗檢測YAP下調對胃癌細胞增殖和轉移的影響.結果 YAP在69.23％的胃腺癌組織中高錶達,與正常胃黏膜(0.33 ±0.15)和胃腺瘤(0.42 ±0.20)比較,YAP mRNA在胃腺癌(1.74±0.46)的錶達水平明顯增高(P＜0.001).體外實驗顯示,YAP下調減少PCNA和MMP-2蛋白的錶達,併抑製胃癌細胞增殖活性和轉移能力.結論 YAP在人胃腺癌組織中高錶達,併且YAP下調抑製胃癌細胞的增殖和轉移.
목적 검측Yes상관단백(YAP)재인위선암、위선류급정상위점막조직중적표체,탐토YAP대위암세포증식화전이적작용급기분자궤제.방법 면역조직화학법화실시정량취합매련반응(Real-time PCR)검측YAP재78례인위선암、39례인위선류급46례정상인위점막조직적표체.이용건립만병독소발협RNA(shRNA)파향YAP기인적위암SGC-7901세포주,Western blot법검측증식세포핵항원(PCNA)화기질금속단백매-2(MMP-2)적단백표체수평.새서람(MTT)비색법화Transwell시험검측YAP하조대위암세포증식화전이적영향.결과 YAP재69.23％적위선암조직중고표체,여정상위점막(0.33 ±0.15)화위선류(0.42 ±0.20)비교,YAP mRNA재위선암(1.74±0.46)적표체수평명현증고(P＜0.001).체외실험현시,YAP하조감소PCNA화MMP-2단백적표체,병억제위암세포증식활성화전이능력.결론 YAP재인위선암조직중고표체,병차YAP하조억제위암세포적증식화전이.
Objective To examine the expression of Yes-associated protein (YAP) in human gastric adenocarcinomas,gastric adenomas and normal gastric mucosa tissues,and further explore the effects and molecular mechanisms of YAP on proliferation and metastasis of gastric cancer cells.Methods The expression of YAP in 78 cases of gastric adenocarcinomas,39 cases of gastric adenomas and 46 cases of normal gastric mucosa tissues was detected by using immunohistochemistry and real-time polymerase chain reaction (PCR).In vitro human gastric cancer SGC-7901 cells were stably transfected with lentivirus-mediated YAP small hairpin RNA (shRNA),and the expression levels of proliferating cell nuclear antigen (PCNA) and metalloproteinase-2 (MMP-2) were detected by using Western blotting.The proliferative activity and metastatic capability of gastric cancer cells were evaluated by methyl thiazol tetrazolium (MTT) and Transwell assays.Results The expression of YAP was observed in 69.23％ gastric adenocarcinoma tissues,and its mRNA expression level in gastric adenocarcinoma (1.74 ± 0.46) was significantly increased as compared with that in normal gastric mucosa (0.33 ± 0.15) and gastric adenoma (0.42 ±0.20).Knockdown of YAP by shRNA could down-regulate the expression of PCNA and MMP-2,and inhibit the proliferation and metastasis of gastric cancer cells.Conclusion YAP is strongly expressed in gastric adenocarcinoma,and knockdown of YAP inhibits proliferation and metastasis of gastric cancer cells.
