不同还原态叶酸和胆碱组合对人结肠腺癌细胞hMLH1和hMSH2转录水平的影响  

Effects of combining different reduced states of folate and choline on transcription of hMLH1 and hMSH2 in human colon adenocarcinoma cells

作　　者：李珍[1,2] 倪娟[1,3] 周滔[1,3] 王晗[1] 汪旭[1,3] 

机构地区：[1]云南师范大学生命科学学院,云南昆明650500 [2]云南白药集团股份有限公司,云南昆明650500 [3]云南师范大学生物能源持续开发与利用教育部工程研究中心,云南昆明650500

出　　处：《癌变．畸变．突变》2015年第3期197-201,206,共6页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：国家自然科学基金(31260268);联合基因生物医药有限公司合作项目

摘　　要：目的:探讨不同还原态叶酸和胆碱对人结肠腺癌细胞DNA错配修复基因h MLH1和h MSH2转录水平的影响。方法:根据受试物在生理和离体情形下维持人基因组结构稳定的最适浓度,以含15、30、120 nmol/L的氧化型叶酸(FA)或还原型叶酸(5-Me THF)与12μmol/L氯化胆碱(CC)组合,以及不同浓度CC(1.5、3μmol/L)与120 nmol/L FA组合的修饰RPMI-1640培养基,对人结肠腺癌细胞株COLO205进行20 d干预培养。以实时荧光定量PCR检测h MLH1和h MSH2 m RNA水平。结果:受试细胞株h MLH1和h MSH2 m RNA水平与FA、5-Me THF和CC浓度呈显著负相关(P<0.01或P<0.05)。30和120 nmol/L浓度下,5-Me THF对上述细胞h MLH1和h MSH2转录水平的影响显著强于同等浓度的FA(P<0.01或P<0.05)。结论:FA、5-Me THF和CC缺乏可上调结肠腺癌细胞h MLH1和h MSH2转录水平,这可能是细胞对叶酸和胆碱缺乏导致的DNA损伤和异常甲基化的应答反应。OBJECTIVE： To explore the effects of folate and choline on human DNA mismatch repair genes h MLH1,h MSH2 expressions in human colonic adenocarcinoma cells. METHODS：Modified RPMI-1640 with the combinations of 12 μmol/L choline chlorine（CC） and 15,30,120 nmol/L of folic acid（FA） or 5-methyltetrahydrofolate（5-Me THF）,FA 120 nmol/L and CC（1.5,3 μmol/L） were set as the intervening culture medium. COLO205 cells were cultured for 20 days. The transcription levels of h MLH1 and h MSH2 were analyzed by real-time fluorescence quantitative PCR. RESULTS：Transcription levels of h MLH1 and h MSH2 were significantly negatively correlated significantly with the concentrations of FA,5-Me THF or CC in tested cells（P〈0.01）. 5-Me THF was more effective on h MLH1 and h MSH2 transcription regulation than same concentration of FA at 30 and 120 nmol/L（P〈0.01 or 0.05）. CONCLUSION：We concluded that FA,5-Me THF and CC deficiency up-regulated the expressions of h MLH1 and h MSH2. This may be the cellular response to DNA damage or abnormal methylation induced by folate and choline deficiency.

关 键 词：氧化型叶酸 5-甲基四氢叶酸 胆碱 HMLH1 HMSH2 

分 类 号：R151[医药卫生—营养与食品卫生学]