前列腺癌细胞DU145同步化诱导的DNA损伤反应通路和P13KAkt通路对凋亡抑制因子基因Apollon（BIRC6）mRNA表达的影响  

Study on the effect of DNA damage Response Pathway and PI3K/Akt Pathway Induced by Syn- chronous Culture of Prostate Cancer Cells DU145 on mRNA Expression of Apollon（BIRC6）

作　　者：彭佑共[1] 杨罗艳[1] 金鹏[1] 吴洪涛[1] 易路[1] 欧阳时锋[1] 

机构地区：[1]中南大学湘雅二医院泌尿外科,湖南长沙410011

出　　处：《医学临床研究》2009年第1期8-12,共5页Journal of Clinical Research

摘　　要：【目的】探讨在前列腺癌细胞DU145同步化培养后引起的DNA损伤反应通路和P13K／AKT通路对凋亡抑制因子（IAP）基因Apollon／BIRC6mRNA表达的影响。同时分析Apollon基因所在染色体是否存在特异性的异常。【方法】采用无血清的饥饿培养法使细胞同步在G。期；用含羞草碱、胸腺嘧啶和噻氨酯哒唑分别使细胞同步在G1期、S期和G2／M期并引起DNA损伤反应，同时加入P13K的特异性抑制剂ly294002以阻止P13K／AKT通路。利用RT—PCR半定量法检测ApollonITIRNA在各个细胞周期相的表达情况。细胞遗传学的常规G式显带法，分析凋亡抑制因子基因所在的染色体是否存在特异性的异常。【结果】含羞草碱同步化的G0／G1期细胞达到了78．04％，胸腺嘧啶同步化的S期细胞达到62．19％，噻氨酯哒唑同步化的G2／M60．5％。Apollon基因所在的染色体2D，存在缺失或者重复，如2p-，2p＋。ApollontuRNA的表达，同步化的G2／M、S期细胞组分别与非同步化细胞组比较差异有显著性（P〈0．01），噻氨酯哒唑＋ly294002组与噻氨酯哒唑相比较差异有显著性（P〈0．01）。【结论】前列腺癌细胞株DU145存在某些染色体的结构和数目异常，可能影响某些基因的表达。药物的同步化激活了DNA损伤反应通路和生存信号通路，再通过P13K／Akt通路，在细胞周期的某个时相调控BIRC6／ApollonmRNA的表达。[Objective]To study the effect of DNA damage response pathway and PI3K/Akt pathway induced by synchronous culture of prostate cancer cells DU145 on mRNA expression of Apollon gene, an apop- tosis inhibitory factor, and to analyze the specific abnormality of the chromosome of Apollon（BIRC6）. [Methods]The starve culture method was used to obtain the cell of Go phase. Mimosine, thymidine and nocodazole were Used respectively to obtain Gl , S and G2/M phase cell, and to cause DNA damage response. The specific inhibitor 1y294002 of PI3K was used to prevent PI3K/AKT pathway. Semi-quantitative RT-PCR method was used to detect the expression of Apollon mRNA at different cell cycle phases. The routine method of cytogenetic G-type Banding was used to analyze the specific abnormality of the chromosome in which apollon（BIRC6） located. [Results]The G0/G1 phase cells synchronized by mimosine was up to 78.04 % ,and the S phase cell by thymidine 62.19% ,and the Gz/M phase cell by nocodazole 60.5%. The chromosome 2p in which Apollon gene located presented deletion and duplication, i.e. 2p- ,2p＋. The expression level of Apollon mRNA in the G2/M and S phase cell group were respectively compared with that in the non-synchronous cell group （p〈0. 01）. The expression of Apollon mRNA in the cell group treated with nocodazole and ly294002 was compared with that in the cell group treated with nocodazole（p〈0.01）. [Conclusion]Synchronizing by the drugs can trigger the DNA damage response pathway and survival signal pathway,and then PI3K/Akt pathway, but not p53 pathway. These pathways may control the expression of BIRC6/Apollon mRNA at a certain phase.

关 键 词：前列腺肿瘤/病理学 DNA损伤 脱噬作用 抑制因子 免疫 RNA 信使 

分 类 号：R737.25[医药卫生—肿瘤]