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细胞内稳态S-腺苷甲硫氨酸浓度对人肝细胞生长因子促肝癌细胞增殖效应中的影响
  • 期刊名称:中华实验外科杂志
  • 时间:0
  • 页码:1832-1834
  • 语言:中文
  • 分类:R735.7[医药卫生—肿瘤;医药卫生—临床医学]
  • 作者机构:[1]武汉大学中南医院普外科,430071
  • 相关基金:基金项目:国家自然科学基金资助项目(30872491/C160402);湖北省自然科学基金资助项目(2009CDB292)
  • 相关项目:S-腺苷蛋氨酸对肝细胞生长因子促肝癌细胞增殖效应的影响
中文摘要:

目的观察体外缺氧条件下肝癌细胞株HepG2中缺氧诱导因子-α(HIF-α)和甲硫氨酸腺苷转移酶2A(MAT2A)的表达,探讨HIF-α在低氧条件下对MAT2A基因表达的调控作用。方法构建人MAT2A启动子真核表达载体质粒,CoCl2化学模拟肿瘤缺氧环境,检测缺氧条件下MAT2A启动子活性,HIF-α和MAT2A在HepG细胞中的共定位、mRNA和蛋白水平的表达。以及小干扰RNA(siRNA)沉默HIF-α后对MAT2A基因表达的影响。结果缺氧24h能使HepG2细胞活性增加到高峰(41.26±2.34),同时MAT2A基因的表达也较常氧时显著升高(P〈0.01),siRNA转染HepG2细胞后能显著下调HIF-α蛋白表达(抑制率90%),并导致MAT2A基因的表达也受到明显抑制。结论缺氧促使HepG2细胞中HIF-α在蛋白水平表达升高,缺氧时HIF-α能上调MAT2A基因的表达水平。

英文摘要:

Objective To observe the expression of hypoixa inducible factor-1α (HIF-α) and me- thionine adenosyltransferase-2A (MAT2A) gene in HepG2 cells under hypoxia in vitro, and explore the regulation of MAT2A gene expression by HIF-1α. Methods Human MAT2A promoter eukaryotic expres- sion vector was constructed. CoC12 was used as a chemical hypoxia-inducible reagent to mimic tumor hy- poxic microenvironment. MAT2A promoter activity, and mRNA and protein expression of HIF-αwere de- tected in the HepG2 cells transfected with RNA interference (RNAi) originated by small interfering RNA (siRNA). The change of MAT2A gene expression was observed after HIF-1α gene silencing. Results Under hypoxia for 24 h, HepG2 cell activity was increased to (41.26 ± 2. 34) , and the mRNA and protein expression levels of MAT2A were up-regulated (P 〈 0. 01 ). After siRNA targeting, the HIF-1 α was down- regulated efficiently in HepG2 cells ( inhibition ratio: 90% ), and MAT2A gene was down-regulated as well. Conclusion Hypoxia can increase protein level of HIF-1α in HepG2 ceils, and HIF-1α up-regu- lates the gene expression of MAT2A.

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