P53、14-3-3σ、miR-365在UVB致HaCaT细胞G2_M期阻滞中的作用.doc

  P53、14-3-3σ、miR-365 在 UVB 致 HaCaT 细胞 G2/M 期阻滞中的作用# 5 10  摘要：目的 研究 30mJ/cm2 的 UVB 照射后，HaCaT 细胞周期的改变及其可能机制。方法 实 验采用人永生化上皮细胞（HaCaT）为研究对象，以波长为 305nm 的 UVB 作为干预因子， 首先观察了 HaCaT 细胞在接受 30mJ/cm2 的 UVB 作用后其细胞周期的变化；其次观察了 UVB 照后不同时间点 p53 蛋白、14-3-3σ蛋白表达的改变情况；最后用 miR-365 高表达的 HCcaT 细胞株分析了 miR-365 在 UVB 所致了的周期阻滞中的可能作用。结果 HaCaT 细胞 在接受 30mJ/cm2 的 UVB 作用后 18h 出现较明显的 G2 期阻滞；p53 蛋白、磷酸化的 p53 以 及 14-3-3σ蛋白在 UVB 照射后均有明显升高； HaCaT 细胞在转染了 miR-365 高表达的载 体之后，14-3-3σ蛋白的表达下降，且在 UVB 照射后无明显改变。结论 30mJ/cm2 的 UVB 15 20 25 30 35 40 照射可诱导 HaCaT 细胞发生 G2 期阻滞，p53、14-3-3σ、miR-365 可能在其中发挥了重要 作用。 关键词：UVB；细胞周期阻滞；miR-365；p53；14-3-3σ 中图分类号：R81 The possible role of P53、14-3-3σ and miR-365 in the HaCaT G2/M cell cycle arrest induced by UVB irradiation GUO Yuanxia1, WANG Yinghui1, ZHOU Meijuan2, DING Zhenhua3 (1. School of Public Health and Tropic Medicine, Southern Medical University, Guangzhou 510515; 2. School of Public Health and Tropic Medicine, Southern Meidcal University,Guangzhou 510515; 3. School of Public Health and Tropic Medicine, Southern Medical University,Guangzhou 510515) Abstract: Objective To study the effect of UVB irradiation on the expression of miR-465 in MEF and analysed its target genes and their functions by bioinformatics analysis. Method experiment using MEFp53+/+ and MEFp53-/- two types of cells, first verify the two cells of the p53 gene and protein expression followed by observation of expression of miR-465 in the two types of cell lines after UVB irradiation in 50J/m2 2h, 4h, 6h, 12h, 24h and then analysed target gene and its functions of miR-465 by bioinformatics. Results P53 in mRNA and protein levels showed a lack of state in MEFp53-/-. miR-465 shown downregulation in MEFp53+/+ at different time points after UVB irradiation and of that was upregulated in MEFp53-/- at all the cooresponding time point. Bioinformatic analysis results show that the different databases miR-465 was jointly predicted target genes, 29 of these target genes involved in positive regulation of cel