中华医学遗传学杂志
中華醫學遺傳學雜誌
중화의학유전학잡지
CHINESE JOURNAL OF MEDICAL GENETICS
2010年
1期
60-65
,共6页
朱亚杰%邱萌%周继陶%刘明%刘素蕊%黄娟%毕锋
硃亞傑%邱萌%週繼陶%劉明%劉素蕊%黃娟%畢鋒
주아걸%구맹%주계도%류명%류소예%황연%필봉
P53蛋白%P21WAF1/Cip1蛋白%Mdm2蛋白%Gadd45α蛋白%胃癌
P53蛋白%P21WAF1/Cip1蛋白%Mdm2蛋白%Gadd45α蛋白%胃癌
P53단백%P21WAF1/Cip1단백%Mdm2단백%Gadd45α단백%위암
protein P53%protein P21WAF1/Cip1%protein Mdm2%protein Gadd45α%gastric cancer
目的 探讨P53蛋白转录调控活性在胃癌发生发展中的作用.方法 应用双荧光素酶报告基因测试(dual-luciferase reporter assay)检测人类胃癌细胞和正常细胞中p53转录调控活性,并通过逆转录-PCR对细胞中p53全长进行扩增,构建pMD18-T-p53重组质粒测序,检测p53突变对其活性的影响.应用免疫组织化学的方法 联合检测P21WAF1/Cip1,Gadd45α,Mdm2三种蛋白在76例胃癌及癌旁组织中表达水平,通过分析它们与肿瘤的临床病理特征及各个分子之间的相关性来间接反应P53在组织中的转录调控活性.结果 双荧光素酶报告基因显示p53转录调控活性在正常细胞中明显高于肿瘤细胞,在p53基因发生突变的细胞系MKN28中P53蛋白转录活性最低.免疫组化结果显示P21WAF1/Cip1,Gadd45α,Mdm2在癌旁组织中的表达明显高于肿瘤组织(P<0.05),且随着肿瘤TNM分期的增高有表达降低的趋势(P<0.05).P21WAF1/Cip1,Gadd45α,Mdm2之间的相关性分析结果显示3个分子在胃癌组织和癌旁组织中的表达呈正相关(P<0.05).结论 P53蛋白转录调控活性的变化在胃癌的发生发展中起着重要的作用,p53突变可以影响其转录调控活性.P21WAF1/Cip1,Gadd45α,Mdm2联合检测可以作为间接反映P53蛋白转录调控活性的效应分子.
目的 探討P53蛋白轉錄調控活性在胃癌髮生髮展中的作用.方法 應用雙熒光素酶報告基因測試(dual-luciferase reporter assay)檢測人類胃癌細胞和正常細胞中p53轉錄調控活性,併通過逆轉錄-PCR對細胞中p53全長進行擴增,構建pMD18-T-p53重組質粒測序,檢測p53突變對其活性的影響.應用免疫組織化學的方法 聯閤檢測P21WAF1/Cip1,Gadd45α,Mdm2三種蛋白在76例胃癌及癌徬組織中錶達水平,通過分析它們與腫瘤的臨床病理特徵及各箇分子之間的相關性來間接反應P53在組織中的轉錄調控活性.結果 雙熒光素酶報告基因顯示p53轉錄調控活性在正常細胞中明顯高于腫瘤細胞,在p53基因髮生突變的細胞繫MKN28中P53蛋白轉錄活性最低.免疫組化結果顯示P21WAF1/Cip1,Gadd45α,Mdm2在癌徬組織中的錶達明顯高于腫瘤組織(P<0.05),且隨著腫瘤TNM分期的增高有錶達降低的趨勢(P<0.05).P21WAF1/Cip1,Gadd45α,Mdm2之間的相關性分析結果顯示3箇分子在胃癌組織和癌徬組織中的錶達呈正相關(P<0.05).結論 P53蛋白轉錄調控活性的變化在胃癌的髮生髮展中起著重要的作用,p53突變可以影響其轉錄調控活性.P21WAF1/Cip1,Gadd45α,Mdm2聯閤檢測可以作為間接反映P53蛋白轉錄調控活性的效應分子.
목적 탐토P53단백전록조공활성재위암발생발전중적작용.방법 응용쌍형광소매보고기인측시(dual-luciferase reporter assay)검측인류위암세포화정상세포중p53전록조공활성,병통과역전록-PCR대세포중p53전장진행확증,구건pMD18-T-p53중조질립측서,검측p53돌변대기활성적영향.응용면역조직화학적방법 연합검측P21WAF1/Cip1,Gadd45α,Mdm2삼충단백재76례위암급암방조직중표체수평,통과분석타문여종류적림상병리특정급각개분자지간적상관성래간접반응P53재조직중적전록조공활성.결과 쌍형광소매보고기인현시p53전록조공활성재정상세포중명현고우종류세포,재p53기인발생돌변적세포계MKN28중P53단백전록활성최저.면역조화결과현시P21WAF1/Cip1,Gadd45α,Mdm2재암방조직중적표체명현고우종류조직(P<0.05),차수착종류TNM분기적증고유표체강저적추세(P<0.05).P21WAF1/Cip1,Gadd45α,Mdm2지간적상관성분석결과현시3개분자재위암조직화암방조직중적표체정정상관(P<0.05).결론 P53단백전록조공활성적변화재위암적발생발전중기착중요적작용,p53돌변가이영향기전록조공활성.P21WAF1/Cip1,Gadd45α,Mdm2연합검측가이작위간접반영P53단백전록조공활성적효응분자.
Objective To investigate the transcriptional activity of P53 in gastric cancer. Methods The activity of p53 in gastric cancer was investigatedby dual-luciferase reporter assay. The coding sequence of the p53 was amplified by reverse transcription-polymerase chain reaction(RT-PCR) and sequenced. The expression of P21WAF1/Cip1, Gadd45α and Mdm2 was detected by immunohistochemistry in 76 samples of gastric carcinoma, and their adjacent tissues were analyzed as control. Results Thep53 activity was higher in human normal cell lines than that of gastric cancer. Furthermore, the transcriptional activity of P53 was lowest in MKN28 cells in which p53 was mutated. The expression level of P21WAF1/Cip1 , Gadd45α and Mdm2 in the adjacent tissues was higher than that of cancer tissues (P<0.05), and there was a tendency of decline in the positive ratio with the poor differentiation of the carcinoma (P<0.05). In addition, a strong linear correlation was observed between P21WAF1/Cip1 , Gadd45α and Mdm2 (P<0.05). Conclusion Changes of P53 transcriptional activity play an important role in the development of gastric cancer. p53 mutation could affect its transcriptional activity. P21WAF1/Cip1 , Gadd45α and Mdm2 are a group of effecters that could reflect P53 transcriptional activity when detected together in cancer tissues.
