中华消化杂志
中華消化雜誌
중화소화잡지
Chinese Journal of Digestion
2009年
5期
322-325
,共4页
左云%高军%李兆申%宋宇%涂建成%程志健%王浩%丰宇芳%陈亚楠%刘苏霞%龚燕芳
左雲%高軍%李兆申%宋宇%塗建成%程誌健%王浩%豐宇芳%陳亞楠%劉囌霞%龔燕芳
좌운%고군%리조신%송우%도건성%정지건%왕호%봉우방%진아남%류소하%공연방
胃肿瘤%Hedgehog蛋白质%甲基化
胃腫瘤%Hedgehog蛋白質%甲基化
위종류%Hedgehog단백질%갑기화
Stomach neoplasms%Hedgehog proteins%Methylation
目的 研究Hedgehog通路中Ptchl基因表达及其甲基化状态在胃癌发生中的变化.方法 分别抽提10例胃癌组织及其癌旁＞3 cm组织和胃癌细胞株AGS的总RNA和基因组DNA.实时定量逆转录多聚酶链反应(QRT-PCR)检测Ptch基因的mRNA表达.应用软件分析Ptchl基因5'调控序列的CpG岛状态,亚硫酸氢盐测序PCR(BSP)分析甲基化水平.结果 对Ptchl mRNAla转录体转录起始位点(计为0点)上游-3950 bp和下游+2050 bp进行CpG岛分析,发现存在两个CpG岛,第1个为-1139 bp～+860 bp,第2个为+875 bp～+1692 bp.以第1个CpG岛的-870 bp～+229 bp区段内的19个CpG位点的BSP测序结果显示,胃癌细胞株AGS全部发生甲基化,胃癌组织中甲基化程度为16%～100%,平均64%±32%,癌旁组织甲基化程度为0%～42%,平均13%±14%,两组问差异有统计学意义(P＜0.05).Spearman秩相关分析发现,Ptchl基因甲基化同其表达呈负相关(r=-0.558,P=0.011).结论 Ptchl基因高甲基化参与胃癌的发生,可能为胃癌新的肿瘤标志物.
目的 研究Hedgehog通路中Ptchl基因錶達及其甲基化狀態在胃癌髮生中的變化.方法 分彆抽提10例胃癌組織及其癌徬＞3 cm組織和胃癌細胞株AGS的總RNA和基因組DNA.實時定量逆轉錄多聚酶鏈反應(QRT-PCR)檢測Ptch基因的mRNA錶達.應用軟件分析Ptchl基因5'調控序列的CpG島狀態,亞硫痠氫鹽測序PCR(BSP)分析甲基化水平.結果 對Ptchl mRNAla轉錄體轉錄起始位點(計為0點)上遊-3950 bp和下遊+2050 bp進行CpG島分析,髮現存在兩箇CpG島,第1箇為-1139 bp～+860 bp,第2箇為+875 bp～+1692 bp.以第1箇CpG島的-870 bp～+229 bp區段內的19箇CpG位點的BSP測序結果顯示,胃癌細胞株AGS全部髮生甲基化,胃癌組織中甲基化程度為16%～100%,平均64%±32%,癌徬組織甲基化程度為0%～42%,平均13%±14%,兩組問差異有統計學意義(P＜0.05).Spearman秩相關分析髮現,Ptchl基因甲基化同其錶達呈負相關(r=-0.558,P=0.011).結論 Ptchl基因高甲基化參與胃癌的髮生,可能為胃癌新的腫瘤標誌物.
목적 연구Hedgehog통로중Ptchl기인표체급기갑기화상태재위암발생중적변화.방법 분별추제10례위암조직급기암방＞3 cm조직화위암세포주AGS적총RNA화기인조DNA.실시정량역전록다취매련반응(QRT-PCR)검측Ptch기인적mRNA표체.응용연건분석Ptchl기인5'조공서렬적CpG도상태,아류산경염측서PCR(BSP)분석갑기화수평.결과 대Ptchl mRNAla전록체전록기시위점(계위0점)상유-3950 bp화하유+2050 bp진행CpG도분석,발현존재량개CpG도,제1개위-1139 bp～+860 bp,제2개위+875 bp～+1692 bp.이제1개CpG도적-870 bp～+229 bp구단내적19개CpG위점적BSP측서결과현시,위암세포주AGS전부발생갑기화,위암조직중갑기화정도위16%～100%,평균64%±32%,암방조직갑기화정도위0%～42%,평균13%±14%,량조문차이유통계학의의(P＜0.05).Spearman질상관분석발현,Ptchl기인갑기화동기표체정부상관(r=-0.558,P=0.011).결론 Ptchl기인고갑기화삼여위암적발생,가능위위암신적종류표지물.
Objective To investigate the expression and aberrant methylation of Ptchl gene in hedgehog signal pathway in carcinogenesis of human gastric cancer.Methods The total RNA and genomic DNA were extracted from 10 human gastric carcinoma tissues,adjacent tissues(＞3 cm from cancerous tissue)and gastric cancer eell line AGS.Ptchl mRNA expression was detected by real-time quantitative reverse transcription PCR.The pattern of CpG island in Ptchl gene 5'regulation sequence was analyzed by software and its methylation extent was tested by bisulfite sequencing PCR.Results The analysis of CpG island(starting-3950 bases upstream of the Ptchl mRNAla transcription start site and ending 2050 bases downstream)revealed that there were two CpG islands in Ptchl gene 5' regulation sequence(first CpG:-1139 bp～+860 bp;second CpG:+875 bp～+1692 bp).Bisulfite sequencing PCR analysis of 19 CpG sites included in the first CpG island(-870 bp～+229 bp)showed that there was methylation present in all cell lines and the average extent of the methylation of these CpG sites was significantly higher in cancerous tissues(64%±32%,ranged 16%～100%)than that in adjacent tissues(13%±14%,ranged 0%～42%,P＜0.05).There was a negative correlation of the Ptchl methylation with its expression.Conclusion The high methylation of Ptchl gene that involves in the carcinogenesis of human gastric carcer will be a new biomarker for gastric carcer.
