中华口腔医学杂志
中華口腔醫學雜誌
중화구강의학잡지
Chinese Journal of Stomatology
2013年
2期
77-80
,共4页
尹小楠%马玉实%杜娟%范志朋
尹小楠%馬玉實%杜娟%範誌朋
윤소남%마옥실%두연%범지붕
癌,鳞状细胞%组蛋白脱甲基酶类%超音刺猬信号
癌,鱗狀細胞%組蛋白脫甲基酶類%超音刺猬信號
암,린상세포%조단백탈갑기매류%초음자위신호
Carcinoma,squamous cell%Histone demethylases%Sonic hedgehog signaling
目的 检测超音刺猬(sonic hedgehog,Shh)信号在头颈部鳞状细胞癌(squamous cell carcinoma,SCC)中是否具有调节组蛋白去甲基化酶相关基因表达的功能.方法 利用人重组SHH-N蛋白或过表达2型突变的平滑基因(mutant 2 smoothened,M2-SMO)在舌SCC细胞系SCC-6激活Shh信号；利用环靶明(cyclopamine)阻断Shh信号0、2、4和8h后收集细胞,采用实时荧光定量反转录PCR(reverse transcription PCR,RT-PCR)在mRNA水平检测组蛋白去甲基化酶相关基因的表达.结果 利用重组SHH-N蛋白激活Shh信号通路后,组蛋白去甲基化酶-赖氨酸特异性去甲基化酶8(lysine-specific demethylase 8,KDM-8)在mRNA水平表达明显升高1.841～3.591倍(P＜0.01)；过表达M2-SMO后KDM-8在mRNA水平表达明显升高1.358～3.013倍(P＜0.05)；而利用环靶明阻断Shh信号通路后KDM-8的表达明显降低25.6％ ～66.6％ (P ＜0.05).结论 在头颈部SCC-6中组蛋白去甲基化酶KDM-8是Shh信号通路的下游基因,其表达受Shh信号分子的正向调控.
目的 檢測超音刺猬(sonic hedgehog,Shh)信號在頭頸部鱗狀細胞癌(squamous cell carcinoma,SCC)中是否具有調節組蛋白去甲基化酶相關基因錶達的功能.方法 利用人重組SHH-N蛋白或過錶達2型突變的平滑基因(mutant 2 smoothened,M2-SMO)在舌SCC細胞繫SCC-6激活Shh信號；利用環靶明(cyclopamine)阻斷Shh信號0、2、4和8h後收集細胞,採用實時熒光定量反轉錄PCR(reverse transcription PCR,RT-PCR)在mRNA水平檢測組蛋白去甲基化酶相關基因的錶達.結果 利用重組SHH-N蛋白激活Shh信號通路後,組蛋白去甲基化酶-賴氨痠特異性去甲基化酶8(lysine-specific demethylase 8,KDM-8)在mRNA水平錶達明顯升高1.841～3.591倍(P＜0.01)；過錶達M2-SMO後KDM-8在mRNA水平錶達明顯升高1.358～3.013倍(P＜0.05)；而利用環靶明阻斷Shh信號通路後KDM-8的錶達明顯降低25.6％ ～66.6％ (P ＜0.05).結論 在頭頸部SCC-6中組蛋白去甲基化酶KDM-8是Shh信號通路的下遊基因,其錶達受Shh信號分子的正嚮調控.
목적 검측초음자위(sonic hedgehog,Shh)신호재두경부린상세포암(squamous cell carcinoma,SCC)중시부구유조절조단백거갑기화매상관기인표체적공능.방법 이용인중조SHH-N단백혹과표체2형돌변적평활기인(mutant 2 smoothened,M2-SMO)재설SCC세포계SCC-6격활Shh신호；이용배파명(cyclopamine)조단Shh신호0、2、4화8h후수집세포,채용실시형광정량반전록PCR(reverse transcription PCR,RT-PCR)재mRNA수평검측조단백거갑기화매상관기인적표체.결과 이용중조SHH-N단백격활Shh신호통로후,조단백거갑기화매-뢰안산특이성거갑기화매8(lysine-specific demethylase 8,KDM-8)재mRNA수평표체명현승고1.841～3.591배(P＜0.01)；과표체M2-SMO후KDM-8재mRNA수평표체명현승고1.358～3.013배(P＜0.05)；이이용배파명조단Shh신호통로후KDM-8적표체명현강저25.6％ ～66.6％ (P ＜0.05).결론 재두경부SCC-6중조단백거갑기화매KDM-8시Shh신호통로적하유기인,기표체수Shh신호분자적정향조공.
Objective To determine whether the sonic hedgehog (Shh) signaling could regulate the expression of histone demethylases in the head and neck squamous cell carcinoma(SCC).Methods Human recombinant SHH-N protein or over-expression of the mutant 2 smoothened (M2-SMO) was applied to activate the Shh signaling in tongue squamous cell carcinoma cell line-SCC-6 in this study.Cyclopamine was used to block the Shh signaling in SCC-6.The real-time reverse transcription (RT)-PCR was used to detect the expression of histone demethylases at the mRNA level.Results The data showed that activation of the Shh signaling up-regulated the expression of histone demethylase,lysine-specific demethylase 8 (KDM-8) at the mRNA level by human recombinant SHH-N protein (1.841 ～ 3.591 fold compare with untreated group;P ＜0.01),over-expression of the M2-SMO also increased the expression of KDM-8 (1.358 ～ 3.013 fold compared with empty vector group ;P ＜ 0.05),and after the Shh signaling was blocked by Cyclopamine,the expression of KDM-8 was down regulated (decreased 25.6％ ～ 66.6％ compared with control cells,P ＜0.05).Conclusions Histone demethylase KDM-8 was downstream target gene of Shh signaling in head and neck squamous cell carcinoma cell line SCC-6,and its expression was positively regulated by the Shh signaling.
