中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2012年
9期
658-663
,共6页
方汉林%于在诚%祝会斌%金永堂
方漢林%于在誠%祝會斌%金永堂
방한림%우재성%축회빈%금영당
5-氮杂-2’-脱氧胞苷%癌,非小细胞肺%SPC-A1细胞%分泌型卷曲相关蛋白1%DNA甲基化
5-氮雜-2’-脫氧胞苷%癌,非小細胞肺%SPC-A1細胞%分泌型捲麯相關蛋白1%DNA甲基化
5-담잡-2’-탈양포감%암,비소세포폐%SPC-A1세포%분비형권곡상관단백1%DNA갑기화
5-Aza-2-deoxycytidine%Carcinoma,non-small cell lung%SPC-A1 cells%Secreted frizzled related protein 1%DNA methylation
目的 观察分泌型卷曲相关蛋白1(SFRP1)基因在非小细胞肺癌(NSCLC)组织中的甲基化状态,研究抑甲基化制剂5-氮杂-2’-脱氧胞苷(5-Aza-CdR)对肺癌SPC-A1细胞中SFRP1、p16和O6-甲基鸟嘌呤-DNA甲基转移酶(MGMT)基因甲基化的影响.方法 采用甲基化特异性PCR(MSP)法和免疫组化SP法,检测60例NSCLC组织中SFRP1基因甲基化的状态和蛋白的表达,以21例肺良性病变组织作为对照.以5-Aza-CdR处理肺癌SPC-A1细胞,采用MSP法和RT-PCR法检测处理前后细胞中 SFRP1、p16和MGMT基因的甲基化状态及相应mRNA的表达.结果 SFRP1基因在60例NSCLC组织中的甲基化率为58.3％,明显高于肺良性病变组织(14.3％;x2 =12.118,P=0.001);SFRP1基因的甲基化与NSCLC的分化程度和淋巴结转移情况有关(均P＜0.05).SFRP1蛋白在35例SFRP1基因甲基化的NSCLC组织中的阴性表达率为68.6％,明显高于非甲基化的NSCLC组织(24.0％；x2=9.613,P=0.002)；SFRP1蛋白的阴性表达与NSCLC的分化程度、临床分期以及淋巴结转移情况有关(均P＜0.05).未用5-Aza-CdR处理时,SPC-A1细胞中SFRP1、p16和MGMT基因甲基化和mRNA的表达量很低；应用不同浓度的5-Aza-dCR处理后,SPC-A1细胞中SFRP1、p16和MGMT基因甲基化和mRNA的表达均显著上调(均P＜0.05).结论 SFRP1基因的甲基化与NSCLC的发生发展密切相关；5-Aza-CdR能逆转SFRP1、p16和MGMT基因的甲基化状态,促进其重新表达.
目的 觀察分泌型捲麯相關蛋白1(SFRP1)基因在非小細胞肺癌(NSCLC)組織中的甲基化狀態,研究抑甲基化製劑5-氮雜-2’-脫氧胞苷(5-Aza-CdR)對肺癌SPC-A1細胞中SFRP1、p16和O6-甲基鳥嘌呤-DNA甲基轉移酶(MGMT)基因甲基化的影響.方法 採用甲基化特異性PCR(MSP)法和免疫組化SP法,檢測60例NSCLC組織中SFRP1基因甲基化的狀態和蛋白的錶達,以21例肺良性病變組織作為對照.以5-Aza-CdR處理肺癌SPC-A1細胞,採用MSP法和RT-PCR法檢測處理前後細胞中 SFRP1、p16和MGMT基因的甲基化狀態及相應mRNA的錶達.結果 SFRP1基因在60例NSCLC組織中的甲基化率為58.3％,明顯高于肺良性病變組織(14.3％;x2 =12.118,P=0.001);SFRP1基因的甲基化與NSCLC的分化程度和淋巴結轉移情況有關(均P＜0.05).SFRP1蛋白在35例SFRP1基因甲基化的NSCLC組織中的陰性錶達率為68.6％,明顯高于非甲基化的NSCLC組織(24.0％；x2=9.613,P=0.002)；SFRP1蛋白的陰性錶達與NSCLC的分化程度、臨床分期以及淋巴結轉移情況有關(均P＜0.05).未用5-Aza-CdR處理時,SPC-A1細胞中SFRP1、p16和MGMT基因甲基化和mRNA的錶達量很低；應用不同濃度的5-Aza-dCR處理後,SPC-A1細胞中SFRP1、p16和MGMT基因甲基化和mRNA的錶達均顯著上調(均P＜0.05).結論 SFRP1基因的甲基化與NSCLC的髮生髮展密切相關；5-Aza-CdR能逆轉SFRP1、p16和MGMT基因的甲基化狀態,促進其重新錶達.
목적 관찰분비형권곡상관단백1(SFRP1)기인재비소세포폐암(NSCLC)조직중적갑기화상태,연구억갑기화제제5-담잡-2’-탈양포감(5-Aza-CdR)대폐암SPC-A1세포중SFRP1、p16화O6-갑기조표령-DNA갑기전이매(MGMT)기인갑기화적영향.방법 채용갑기화특이성PCR(MSP)법화면역조화SP법,검측60례NSCLC조직중SFRP1기인갑기화적상태화단백적표체,이21례폐량성병변조직작위대조.이5-Aza-CdR처리폐암SPC-A1세포,채용MSP법화RT-PCR법검측처리전후세포중 SFRP1、p16화MGMT기인적갑기화상태급상응mRNA적표체.결과 SFRP1기인재60례NSCLC조직중적갑기화솔위58.3％,명현고우폐량성병변조직(14.3％;x2 =12.118,P=0.001);SFRP1기인적갑기화여NSCLC적분화정도화림파결전이정황유관(균P＜0.05).SFRP1단백재35례SFRP1기인갑기화적NSCLC조직중적음성표체솔위68.6％,명현고우비갑기화적NSCLC조직(24.0％；x2=9.613,P=0.002)；SFRP1단백적음성표체여NSCLC적분화정도、림상분기이급림파결전이정황유관(균P＜0.05).미용5-Aza-CdR처리시,SPC-A1세포중SFRP1、p16화MGMT기인갑기화화mRNA적표체량흔저；응용불동농도적5-Aza-dCR처리후,SPC-A1세포중SFRP1、p16화MGMT기인갑기화화mRNA적표체균현저상조(균P＜0.05).결론 SFRP1기인적갑기화여NSCLC적발생발전밀절상관；5-Aza-CdR능역전SFRP1、p16화MGMT기인적갑기화상태,촉진기중신표체.
Objective To observe the expression of SFRP1 gene methylation in non-small cell lung cancer (NSCLC),and study the effect of 5-Aza-2-deoxycytidine (5-Aza-CdR) on DNA methylation and expression of SFRP1,p16 and MGMT genes in the human lung cancer cell line SPC-A-1 cells.Methods SP immunohistochemistry and methylation-specific PCR were used to detect the SFRP1 methylation in 60 NSCLC cases,and 21 cases of benign lung diseases were used as control group.SPC-A-1 cells were cultured and treated with 5-Aza-CdR.The promoter methylation status of SFRP1,p16 and MGMT genes were detected by methylation-specific polymerase (MSP) chain reaction,and mRNAs were detected by real-time PCR.Results The positive rate of SFRP1 gene methylation in NSCLC was significantly higher than that in normal lung tissue (58.3％ vs.14.3％ ; x2 =12.118,P =0.001).SFRP1 gene methylation was closely correlated with lymph node metastasis and degree of differentiation in NSCLC (P ＜ 0.05).SFRP1 protein expression was correlated with clinical stage,degree of differentiation and lymph node metastasis in NSCLC (P ＜0.05).The positive expression of SFRP1 protein in 30 cases of NSCLC tissue containing SFRP1 gene methylation was significantly higher than that in non-methylated NSCLC (68.6％ vs.24.0％ ; x2 =9.613,P =0.002).SFRP1 gene methylation was closely correlated with SFRP1 gene protein expression in NSCLC (P ＜0.05).Negative expression of SFRP1 protein was correlated with the differentiation,clinical stage,and lymph node metastasis in NSCLC (all P ＜ 0.05).Without 5-Aza-CdR treatment,the expressions of methylation of SFRP1,p16 and MGMT genes and their mRNA were low.After 5-Aza-CdR treatment at different concentrations,their expressions were significantly elevated (all P ＜ 0.05).Conclusions SFRP1 gene methylation is closely associated with carcinogenesis and development of NSCLC.5-Aza-CdR may reverse the methylation of SFRP1,p16 and MGMT genes,and facilitate the re-expression of the anti-oncogenes.
