中华胃肠外科杂志
中華胃腸外科雜誌
중화위장외과잡지
CHINESE JOURNAL OF GASTROINTESTINAL SURGERY
2010年
6期
440-444
,共5页
叶青%冯波%彭远飞%蔡劬%陈雪华%俞焙秦%马君俊%陆爱国%李健文%王明亮%刘炳亚%郑民华
葉青%馮波%彭遠飛%蔡劬%陳雪華%俞焙秦%馬君俊%陸愛國%李健文%王明亮%劉炳亞%鄭民華
협청%풍파%팽원비%채구%진설화%유배진%마군준%륙애국%리건문%왕명량%류병아%정민화
结直肠肿瘤%γ-synuclein%甲基化%5-杂氮-2'-脱氧胞苷
結直腸腫瘤%γ-synuclein%甲基化%5-雜氮-2'-脫氧胞苷
결직장종류%γ-synuclein%갑기화%5-잡담-2'-탈양포감
Colorectal neoplasms%γ-synuclein%Methylation%5-aza-2'-deoxycytidine
目的 探讨结直肠癌中γ-synuclein基因的表达与启动子区CpG岛甲基化修饰之间的关系,以及γ-synuclein基因去甲基化水平与结直肠癌临床病理特征的关系.方法 采用半定量RT-PCR法检测30例结直肠癌和相应癌旁组织中γ-synuclein基因的表达及去甲基化试剂5-杂氮-2'-脱氧胞苷(5-Aza-C)干预对结直肠癌细胞COLO205、LoVo和SW480的γ-synuclein基因表达的影响.采用亚硫酸氢盐修饰测序(BSP)法检测5-Aza-C处理前后CpG岛的甲基化情况.采用巢式甲基化PCR(NMSP)及实时荧光定量MSP法检测67例结直肠癌组织和30例相应癌旁组织中γ-synuclein基因的甲基化状态,并分析其与临床病理特征的关系.结果结直肠癌组织中γ-synuclein mRNA的表达水平0.66±0.34,高于相应癌旁组织的0.45±0.26,差异有统计学意义(P=0.011).5-Aza-C处理后,COLO205、LoVo和SW480细胞γ-synuclein mRNA表达水平明显上升,同时γ-synuclein基因启动子区CpG岛的甲基化水平明显降低.80.0%(24/30)的结直肠癌组织和50.0%(15/30)的相应癌旁组织的γ-synuclein基因被去甲基化,结直肠癌组织中γ-synuclein基因去甲基化的趋势明显强于相应癌旁组织(P=0.030).γ-synuclein基因的去甲基化水平与结直肠癌的淋巴结转移、临床病理分期及远处转移有关.结论 结直肠癌中γ-synuclein基因的上调表达与启动子区CpG岛的去甲基化状态有一定关联,γ-synuclein基因的去甲基化水平有望成为结直肠癌患者判断预后的潜在标志物.
目的 探討結直腸癌中γ-synuclein基因的錶達與啟動子區CpG島甲基化脩飾之間的關繫,以及γ-synuclein基因去甲基化水平與結直腸癌臨床病理特徵的關繫.方法 採用半定量RT-PCR法檢測30例結直腸癌和相應癌徬組織中γ-synuclein基因的錶達及去甲基化試劑5-雜氮-2'-脫氧胞苷(5-Aza-C)榦預對結直腸癌細胞COLO205、LoVo和SW480的γ-synuclein基因錶達的影響.採用亞硫痠氫鹽脩飾測序(BSP)法檢測5-Aza-C處理前後CpG島的甲基化情況.採用巢式甲基化PCR(NMSP)及實時熒光定量MSP法檢測67例結直腸癌組織和30例相應癌徬組織中γ-synuclein基因的甲基化狀態,併分析其與臨床病理特徵的關繫.結果結直腸癌組織中γ-synuclein mRNA的錶達水平0.66±0.34,高于相應癌徬組織的0.45±0.26,差異有統計學意義(P=0.011).5-Aza-C處理後,COLO205、LoVo和SW480細胞γ-synuclein mRNA錶達水平明顯上升,同時γ-synuclein基因啟動子區CpG島的甲基化水平明顯降低.80.0%(24/30)的結直腸癌組織和50.0%(15/30)的相應癌徬組織的γ-synuclein基因被去甲基化,結直腸癌組織中γ-synuclein基因去甲基化的趨勢明顯彊于相應癌徬組織(P=0.030).γ-synuclein基因的去甲基化水平與結直腸癌的淋巴結轉移、臨床病理分期及遠處轉移有關.結論 結直腸癌中γ-synuclein基因的上調錶達與啟動子區CpG島的去甲基化狀態有一定關聯,γ-synuclein基因的去甲基化水平有望成為結直腸癌患者判斷預後的潛在標誌物.
목적 탐토결직장암중γ-synuclein기인적표체여계동자구CpG도갑기화수식지간적관계,이급γ-synuclein기인거갑기화수평여결직장암림상병리특정적관계.방법 채용반정량RT-PCR법검측30례결직장암화상응암방조직중γ-synuclein기인적표체급거갑기화시제5-잡담-2'-탈양포감(5-Aza-C)간예대결직장암세포COLO205、LoVo화SW480적γ-synuclein기인표체적영향.채용아류산경염수식측서(BSP)법검측5-Aza-C처리전후CpG도적갑기화정황.채용소식갑기화PCR(NMSP)급실시형광정량MSP법검측67례결직장암조직화30례상응암방조직중γ-synuclein기인적갑기화상태,병분석기여림상병리특정적관계.결과결직장암조직중γ-synuclein mRNA적표체수평0.66±0.34,고우상응암방조직적0.45±0.26,차이유통계학의의(P=0.011).5-Aza-C처리후,COLO205、LoVo화SW480세포γ-synuclein mRNA표체수평명현상승,동시γ-synuclein기인계동자구CpG도적갑기화수평명현강저.80.0%(24/30)적결직장암조직화50.0%(15/30)적상응암방조직적γ-synuclein기인피거갑기화,결직장암조직중γ-synuclein기인거갑기화적추세명현강우상응암방조직(P=0.030).γ-synuclein기인적거갑기화수평여결직장암적림파결전이、림상병리분기급원처전이유관.결론 결직장암중γ-synuclein기인적상조표체여계동자구CpG도적거갑기화상태유일정관련,γ-synuclein기인적거갑기화수평유망성위결직장암환자판단예후적잠재표지물.
Objective To explore the relationship between γ-synuclein gene expression and CpG island demethylation in colorectal cancer (CRC), and the relationship between the demethylation and clinicopathological factors of CRC. Methods The expression of γ-synuclein mRNA was examined in 30 pairs of tumor tissues and tumor-matched non-neoplastic adjacent tissues (NNAT) by RT-PCR.CRC cell lines including COLO205, LoVo, and SW480 were used and treated with a demethylating agent, 5-aza-2'-deoxycytidine (5-aza-C). Before and after the treatment, the expression of γ-synuclein mRNA in the cells was determined by RT-PCR, and bisulfite sequencing PCR was also used to analyze methylation status of CpG island. The methylation status of γ-synuclein was then examined in 67 CRC samples and 30 NNAT samples by nested methylation-specific PCR (NMSP) and real time methylationspecific PCR (real-time MSP). The relationship between the demethylation of γ-synuclein in CRC and clinicopathological factors was analyzed. Results The mean γ-synuclein mRNA expression was 0.66±0.34 in CRC samples, which was much higher than 0.45±0.26 in NNAT samples (P=0.011). 5-aza-C could induce expression and demethylation of γ-synuclein in COLO205, LoVo and SW480 cells. Γ-Synuclein gene was demethylated in 80.0%(24/30) of the CRC samples and 50.0%(15/30) of the NNAT samples.The demethylated status of γ-synuclein was much higher in CRC samples than that in NNAT samples (P=0.030), and was significantly correlated with clinical stage, lymph node involvement, and distant metastasis of CRC (P＜0.05). Conclusion The upregulation of γ-synuclein expression in CRC is primarily attributed to the demethylation of CpG island, which may be used as a marker for prognosis.
