中华普通外科杂志
中華普通外科雜誌
중화보통외과잡지
CHINESE JOURNAL OF GENERAL SURGERY
2008年
9期
702-705
,共4页
王歆光%王维民%杨尹默%刘玉村%万远廉
王歆光%王維民%楊尹默%劉玉村%萬遠廉
왕흠광%왕유민%양윤묵%류옥촌%만원렴
癌%肝细胞%基因表达%环耙明%Hedgehog
癌%肝細胞%基因錶達%環耙明%Hedgehog
암%간세포%기인표체%배파명%Hedgehog
Carcinoma,hepatocellular%Gene expression%Cyclopamine%Hedgehog
目的 研究Hedgehog信号传导通路相关基因在肝癌组织中的表达,及其抑制剂对肿瘤细胞生长的影响.方法 免疫组化法检测14例肝癌组织切片、4个肝癌细胞系和正常肝细胞中Hedgehog信号传导通路成员Ihh、Ptch、Smo、Gli的蛋白表达;Western blot检测9例新鲜肝癌组织标本、6例正常肝组织标本和肝癌细胞系中Ihh、Ptch、Smo、Gh的蛋白表达;RT-PCR检测3肝癌组织标本和肝癌细胞系中Ihh、Ptch、Smo、Gli、Hip mRNA表达.结果 14例肝癌组织中免疫组化染色阳性的Gli 6例(42.9%),Ptch 10例(71.4%),Ihh 10例(71.4%),Smo 12例(85.7%).Western blot和RT-PCR检测显示,肿瘤组织中Gli基因的mRNA和蛋白表达均高于正常肝脏细胞,Hip基因的mRNA表达低于正常肝细胞.肝癌细胞系HepG2,Bel-7402和QGY-7701免疫细胞化学染色Pteh和Gli同时为阳性,证明为Hedgehog通路活化的细胞系.在环耙明(KAAD-cyclopamine)作用后,3个细胞系Ptch及Gli基因的mRNA表达下调(Ptch基因tHepG2=3.78,tBel-7402=9.03,tQGY-7701=5.63,Gli基因tHepG2=9.61,tBel-7402=4.15,tQGY-7701=20.30,均P＜0.05);QGY-7701组Hip基因RNA表达上调(t=4.70,P＜0.05).环耙明抑制肝癌细胞系QGY-7701的作用最明显.结论 原发性肝癌组织中Hedgehog信号传导通路有多种基因表达,环耙明有抑制肝癌细胞生长的作用.
目的 研究Hedgehog信號傳導通路相關基因在肝癌組織中的錶達,及其抑製劑對腫瘤細胞生長的影響.方法 免疫組化法檢測14例肝癌組織切片、4箇肝癌細胞繫和正常肝細胞中Hedgehog信號傳導通路成員Ihh、Ptch、Smo、Gli的蛋白錶達;Western blot檢測9例新鮮肝癌組織標本、6例正常肝組織標本和肝癌細胞繫中Ihh、Ptch、Smo、Gh的蛋白錶達;RT-PCR檢測3肝癌組織標本和肝癌細胞繫中Ihh、Ptch、Smo、Gli、Hip mRNA錶達.結果 14例肝癌組織中免疫組化染色暘性的Gli 6例(42.9%),Ptch 10例(71.4%),Ihh 10例(71.4%),Smo 12例(85.7%).Western blot和RT-PCR檢測顯示,腫瘤組織中Gli基因的mRNA和蛋白錶達均高于正常肝髒細胞,Hip基因的mRNA錶達低于正常肝細胞.肝癌細胞繫HepG2,Bel-7402和QGY-7701免疫細胞化學染色Pteh和Gli同時為暘性,證明為Hedgehog通路活化的細胞繫.在環耙明(KAAD-cyclopamine)作用後,3箇細胞繫Ptch及Gli基因的mRNA錶達下調(Ptch基因tHepG2=3.78,tBel-7402=9.03,tQGY-7701=5.63,Gli基因tHepG2=9.61,tBel-7402=4.15,tQGY-7701=20.30,均P＜0.05);QGY-7701組Hip基因RNA錶達上調(t=4.70,P＜0.05).環耙明抑製肝癌細胞繫QGY-7701的作用最明顯.結論 原髮性肝癌組織中Hedgehog信號傳導通路有多種基因錶達,環耙明有抑製肝癌細胞生長的作用.
목적 연구Hedgehog신호전도통로상관기인재간암조직중적표체,급기억제제대종류세포생장적영향.방법 면역조화법검측14례간암조직절편、4개간암세포계화정상간세포중Hedgehog신호전도통로성원Ihh、Ptch、Smo、Gli적단백표체;Western blot검측9례신선간암조직표본、6례정상간조직표본화간암세포계중Ihh、Ptch、Smo、Gh적단백표체;RT-PCR검측3간암조직표본화간암세포계중Ihh、Ptch、Smo、Gli、Hip mRNA표체.결과 14례간암조직중면역조화염색양성적Gli 6례(42.9%),Ptch 10례(71.4%),Ihh 10례(71.4%),Smo 12례(85.7%).Western blot화RT-PCR검측현시,종류조직중Gli기인적mRNA화단백표체균고우정상간장세포,Hip기인적mRNA표체저우정상간세포.간암세포계HepG2,Bel-7402화QGY-7701면역세포화학염색Pteh화Gli동시위양성,증명위Hedgehog통로활화적세포계.재배파명(KAAD-cyclopamine)작용후,3개세포계Ptch급Gli기인적mRNA표체하조(Ptch기인tHepG2=3.78,tBel-7402=9.03,tQGY-7701=5.63,Gli기인tHepG2=9.61,tBel-7402=4.15,tQGY-7701=20.30,균P＜0.05);QGY-7701조Hip기인RNA표체상조(t=4.70,P＜0.05).배파명억제간암세포계QGY-7701적작용최명현.결론 원발성간암조직중Hedgehog신호전도통로유다충기인표체,배파명유억제간암세포생장적작용.
Objective To investigate the expressions of Hedgehog sisnaling pathway genes in hepatocellular carcinoma tissues(HCC),and the effect of specific Hedgehog pathway inhibitor(KAADcyclopamine)on the growth of HCC cells and the expressions of Hedgehog genes. Methods The expression of Hedgehog signaling pathway components(Ihh,Ptch,Smo and Gli)was investigated in 14 HCC tissue slices,4 HCC cell lines and a normal hepatic cell line by using immunochemistry.The expression of Ihh,Ptch,Smo and Gli proteins was investigated in 9 HCC tissue specimens and 6 normal hepatic tissue specimens by using Western blotting.The expression of Ihh、Ptch、Smo、Gli and Hip genes was investigated by RT-PCR.Results The positive ratio of Gli,Ptch,Ihh and Smo were 42.9%,71.4%,71.4% and 85.7% in 14 HCC tissue slices,respectively.The expressions of Gli protein and Gii gene were up regulated while the expression of Hip gene was down regulated in HCC specimens compared with normal hepatic tissue specimens.Hedgehog signaling pathways in HCC cell lines HepG2,Bel-7402 and QGY-7701 were activated;KAAD-cyclopamine,a specific inhibitor of the Hedgehog signaling pathway,down regulated cell growth and the expressions of Ptch and Gli genes in the 3 HCC cell lines(Ptch gene:tHepG2=3.78,tBel-7402=9.03,tQGY-7701=5.63;Gli gene:tHepG2=9.61,tBel-7402=4.15,tQGY-7701=20.30,P＜0.05 in each group).The expression of Hip gene was up regulated in QGY-7701 after treated with KAAD-cyclopamine(t=4.70,P＜0.05).Conclusion The expression of main Hedgehog signaling pathway components were detected in HCC,KAAD-cyclopamine specifically inhibited the Hedgehog signaling pathway.
