| 引用本文： |
-
徐成党,黄盛松,钱多成,等.DNA羟甲基化酶TET1对良性前列腺增生细胞的增殖调控作用及其机制[J].同济大学学报（医学版）,2019,40(1):10-15. [点击复制]
- XU Cheng-dang,HUANG Sheng-song,QIAN Duo-cheng,et al.Effect of DNA hydroxymethylase TET1 on proliferation of benign prostatic hyperplasia cells and its mechanisms[J].同济大学学报（医学版）,2019,40(1):10-15. [点击复制]
|
|
| 摘要: |
| 目的 研究DNA羟甲基化酶TET1在前列腺增生细胞增殖调控中的作用及其机制。方法 按前列腺体积将患者分成3组： A组<30mL、30mL≤B组≤70mL、C组>70mL，收集手术前列腺外周带和移行带标本，用酶联免疫吸附实验方法比较检测两者双氢睾酮（DHT）的含量差异，免疫组化检测比较两者TET1、Ⅱ型5α-还原酶和甲基化的表达差异。采用慢病毒转染方法过表达和敲减正常前列腺上皮细胞RWPE-1中TET1基因后，CCK-8检测基因过表达或敲减和对照组细胞的增殖情况，流式细胞术检侧各组细胞凋亡，实时聚合酶链反应和Western印迹法检测各组Ⅱ型5α-还原酶表达水平。结果 B和C组移行带中DHT含量高于外周带（P<0.05），移行带中TET1和Ⅱ型5α-还原酶表达高于外周带，甲基化水平低于外周带（P<0.05），A组中无明显差异（P>0.05）。与对照组TET1-Ctrl（0.98±0.22）相比，TET1-OE组Ⅱ型5α-还原酶蛋白表达量（1.52±0.34）增加（P<0.05），SRD5A2表达升高（P<0.05），细胞凋亡水平下降（P<0.05），TET1-KD组结果则相反。结论 在前列腺增生过程中，可能是DNA羟甲基化酶TET1发挥去甲基化作用而降低移行带中Ⅱ型5α-还原酶基因SRD5A2启动子的甲基化水平，Ⅱ型5α-还原酶表达增加促进睾酮转化为DHT，移行带细胞增殖凋亡平衡被打破而导致过度增殖。 |
| 关键词: 前列腺增生 TET1基因 Ⅱ型5α-还原酶 甲基化 |
| DOI：10.16118/j.1008-0392.2019.01.003 |
| 投稿时间：2018-06-04 |
| 基金项目:上海市教委科研创新项目(15ZZ022) |
|
| Effect of DNA hydroxymethylase TET1 on proliferation of benign prostatic hyperplasia cells and its mechanisms |
| XU Cheng-dang,HUANG Sheng-song,QIAN Duo-cheng,WU Deng-long |
| (Dept. of Urology, Tongji Hospital, Tongji University, Shanghai 200065, China) |
| Abstract: |
| Objective To investigate the role and mechanism of DNA hydroxymethylase TET1 in the proliferation regulation of prostate hyperplasia cells. Methods Seventy-two patients with benign prostatic hyperplasia(BPH) were divided into three groups according to the prostate volume： group A(<30mL, n=11), group B(≥30mL,≤70mL, n=34) and group C(>70mL, n=27). The surgical specimens of prostate peripheral zone and transition zone were collected, the dihydrotestosterone(DHT) levels were measured with ELISA, the expression of methylation, TET1 and 5 α-reductase Ⅱ was detected with immunohistochemistry. The TET1 gene of normal prostate epithelial cells RWPE-1 was knocked down by transfection with lentivirus vector; the cell proliferation, expression of protein and mRNA of 5 α-reductase Ⅱ, apoptosis were examined with CCK-8,Western blotting,qPCR and the flow cytometry, respectively. Results The DHT level and the expression of TET1 and 5 α-reductase Ⅱ of the transitional zone were higher; the methylation level was lower than those of peripheral zone in group B and C(all P<0.05); however, there were no significant differences in group A(P>0.05). Compared to RWPE-1 control cells(TET1-Ctr), the expression of 5 α-reductase Ⅱ in TET1 over-expressing RWPE-1 cells(TET1-OE）was increased(1.52±0.34 vs 0.98±0.22, P<0.05), and SRD5A2 expression also increased（P<0.05）and the level of apoptosis decreased(P<0.05); the results of TET1 knockdown RWPE-1 cells(TET1-KD) were opposite. Conclusion The study indicates that in the process of hyperplasia of prostate, the hydroxyl DNA methylation of TET1 may induce the level of methylation of the SRD5A2 gene in the transitional zone, then increase the expression of 5 α-reductase Ⅱ to enhance the conversion of testosterone to DHT, resulting the proliferation and apoptosis balance broken of the transitional zone and cell excessive proliferation. |
| Key words: prostate hyperplasia TET1 genes 5 α-reductase Ⅱ methylation |
