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

doi:10.3969/j.issn.1004-616x.2015.03.008

癌变·畸变·突变 ›› 2015, Vol. 27 ›› Issue (3): 197-201,206.doi: 10.3969/j.issn.1004-616x.2015.03.008

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

李珍^1,3, 倪娟^1,2, 周滔^1,2, 王晗¹, 汪旭^1,2

1. 云南师范大学生命科学学院, 云南昆明 650500;
2. 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;
3. 云南白药集团股份有限公司, 云南昆明 650500

收稿日期:2014-12-09 修回日期:2015-04-22 出版日期:2015-05-30 发布日期:2015-05-30
通讯作者: 汪旭,E-mail:wangxu@fudan.edu.cn E-mail:wangxu@fudan.edu.cn
作者简介:李珍,E-mail:lizhen06jibban@126.com
基金资助:
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

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

LI Zhen^1,3, NI Juan^1,2, ZHOU Tao^1,2, WANG Han¹, WANG Xu^1,2

1. School of Life Sciences, Yunnan Normal University, Kunming 650500;
2. The Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University, Kunming 650500;
3. Yunnan Baiyao Group Co., Ltd., Kunming 650500, Yunnan, China

Received:2014-12-09 Revised:2015-04-22 Online:2015-05-30 Published:2015-05-30

摘要/Abstract

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hMLH1和hMSH2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-MeTHF)与12 μmol/L氯化胆碱(CC)组合,以及不同浓度CC(1.5、3 μmol/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hMLH1和hMSH2 mRNA水平。结果:受试细胞株hMLH1和hMSH2 mRNA水平与FA、5-MeTHF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-MeTHF对上述细胞hMLH1和hMSH2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-MeTHF和CC缺乏可上调结肠腺癌细胞hMLH1和hMSH2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。

关键词: 氧化型叶酸, 5-甲基四氢叶酸, 胆碱, hMLH1, hMSH2

Abstract:

OBJECTIVE: To explore the effects of folate and choline on human DNA mismatch repair genes hMLH1, hMSH2 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-MeTHF), 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 hMLH1 and hMSH2 were analyzed by real-time fluorescence quantitative PCR. RESULTS:Transcription levels of hMLH1 and hMSH2 were significantly negatively correlated significantly with the concentrations of FA, 5-MeTHF or CC in tested cells (P<0.01). 5-MeTHF was more effective on hMLH1 and hMSH2 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-MeTHF and CC deficiency up-regulated the expressions of hMLH1 and hMSH2. This may be the cellular response to DNA damage or abnormal methylation induced by folate and choline deficiency.

Key words: folic acid, 5-methyltetrahydrofolate, choline, hMLH1, hMSH2

中图分类号:

R151

李珍, 倪娟, 周滔, 王晗, 汪旭. 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[J]. 癌变·畸变·突变, 2015, 27(3): 197-201,206.

LI Zhen, NI Juan, ZHOU Tao, WANG Han, WANG Xu. Effects of combining different reduced states of folate and choline on transcription of hMLH1 and hMSH2 in human colon adenocarcinoma cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2015, 27(3): 197-201,206.

参考文献

[1] Bevilacqua RA, Simpson AJ. Methylation of the hMLH1 promoter but no hMLH1 mutations in sporadic gastric carcinomas with high-level microsatellite instability[J]. Int J Cancer, 2000, 87(2):200-203.
[2] Kamory E, Kolacsek O, Otto S, et al. hMLH1 and hMSH2 somatic inactivation mechanisms in sporadic colorectal cancer patients[J]. Pathol Oncol Res, 2003, 9(4):236-241.
[3] Zhang H, Zhang S, Cui J, et al. Expression and promoter methylation status of mismatch repair gene hMLH1 and hMSH2 in epithelial ovarian cancer[J]. Aust NZJ Obstet Gynaecol, 2008, 48(5):505-509.
[4] 汪旭, 薛京伦. 叶酸代谢与人类基因组稳定性的关系研究进展[J]. 国外医学:遗传学分册, 2005, 28(5):257-261.
[5] Ross SA. Diet and DNA methylation interactions in cancer prevention[J]. Ann NY Acad Sci, 2003, 983:197-207.
[6] Kimura M, Umegaki K, Higuchi M, et al. Methylenetetra-hydrofolate reductase C677T polymorphism, folic acid and riboflavin are important determinants of genome stability in cultured human lymphocytes[J]. J Nutr, 2004, 134(1):48-56.
[7] da Costa KA, Niculescu MD, Craciunescu CN, et al. Choline deficiency increases lymphocyte apoptosis and DNA damage in humans[J]. Am J Clin Nutr, 2006, 84(1):88-94.
[8] Fenech M. Folate, DNA damage and the aging brain[J]. Mech Ageing Dev, 2010, 131(4):236-241.
[9] Duthie SJ. Folate and cancer:how DNA damage, repair and methylation impact on colon carcinogenesis[J]. J Inherit Metab Dis, 2011, 34(1):101-109.
[10] 鲁琳, 高微微, 倪娟, 等. 胆碱与叶酸组合对人成淋巴细胞系DNA损伤效应[J]. 营养学报, 2010, 32(1):26-29.
[11] Wang X, Fenech M. A comparison of folic acid and 5-methyltetrahydrofolate for prevention of DNA damage and cell death in human lymphocytes in vitro[J]. Mutagenesis, 2003, 18(1):81-86.
[12] Fenech M. The role of folic acid and Vitamin B12 in genomic stability of human cells[J]. Mutat Res, 2001, 475(1/2):57-67.
[13] Vageli D, Daniil Z, Dahabreh J, et al. Phenotypic mismatch repair hMSH2 and hMLH1 gene expression profiles in primary non-small cell lung carcinomas[J]. Lung Cancer, 2009, 64 (3):282-288.
[14] 寸燕萍, 倪娟, 徐伟江, 等. 氧化型/还原型叶酸不足对人淋巴细胞hMSH2转录的影响[J]. 肿瘤防治研究杂志, 2011, 38(11):1320-1321.
[15] Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT Method[J]. Methods, 2001, 25(4):402-408.
[16] Fenech M. Genome health nutrigenomics and nutrigenetics diagnosis and nutritional treatment of genome damage on an individual basis[J]. Food Chem Toxicol, 2008, 46(4):1365-1370.
[17] Zeisel SH. Importance of methyl donors during reproduction[J]. Am J Clin Nutr, 2009, 89(2):673-677.
[18] Maeda T, Guan JZ, Oyama J, et al. Age-related changes in subtelomeric methylation in the normal Japanese population[J]. J Gerontol A Biol Sci Med Sci, 2009, 64(4):426-434.
[19] Jones PA, Baylin SB. The fundamental role of epigenetic events in cancer[J]. Nat Rev Genet, 2002, 3(6):415-428.

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

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 7

编辑推荐

Metrics

本文评价

[1]	徐海明, 郝艳星, 陈娅楠, 杨惠芳, 李江平, 朱玲勤, 任彬彬, 德小明. 羟胺三氯化铁法测定人血清胆碱酯酶方法的优化[J]. 癌变·畸变·突变, 2020, 32(2): 145-148,154.
[2]	赵敏娴, 姚欣雅, 曹正颖, 王灿楠. 大鼠一次性经皮染毒毒死蜱的毒代动力学和毒效应学研究[J]. 癌变·畸变·突变, 2015, 27(2): 129-133,145.
[3]	吴惠,乙楠楠,李亭亭,何贤松,赵敏娴,姚欣雅,王灿楠. 毒死蜱和乐果联合染毒对大鼠胆碱酯酶活性的影响[J]. 癌变·畸变·突变, 2013, 25(1): 31-34.
[4]	高微微;徐伟江;周滔;汪旭. 胆碱与叶酸不同浓度组合诱导正常人与乳腺癌患者成淋巴细胞凋亡效应的比较[J]. , 2010, 22(3): 210-213.
[5]	鲁琳;高微微;徐伟江;周滔;汪旭. 胆碱与叶酸不足对人成淋巴细胞株细胞死亡的影响[J]. , 2010, 22(2): 123-125.
[6]	朱　欣1;连灵君1;王　燕1;刘慧刚1;徐立红1,2. 三丁基锡暴露可使大鼠脑乙酰胆碱酯酶活性升高[J]. , 2006, 18(4): 292-293.
[7]	和智君　李晓琼　汪　旭. 胆碱受体拮抗剂在小鼠骨髓细胞中的C- 有丝分裂效应研究[J]. , 1999, 11(4): 165-167.