CDADC1在人视网膜色素上皮细胞中功能研究

目的：研究细胞增殖相关基因CDADC1在人视网膜色素上皮细胞ARPE19中的表达及对ARPE19细胞增殖的影响。方法：采用基因重组技术构建荧光表达载体pEGFP－C1－CDADC1和真核表达载体pcDNA3．1-myc-CDADC1，用脂质体转染法转染ARPE19细胞，观察GFP—CDADC1融合蛋白在ARPE19细胞的表达定位，流式细胞仪测定CDADC1转染后对ARPE19细胞生长周期、凋亡的影响。结果：FP—CDADC1融合蛋白亚细胞定位显示，CDADC1低表达于ARPE19细胞胞浆，高表达于细胞核；pcD—NA3．1-myc-CDADC1瞬时转染ARPE19细胞显示24小时细胞无明显改变，48小时后重组质粒转染组S期细胞占细胞总数的19．37％，pcDNA3．1-myc空载质粒转染组S期细胞占10．87％，而空白对照组S期细胞占3．33％，重组质粒转染组与两对照组之间的差异有统计学意义（P〈0．01）。结论：CDADC1在增生性玻璃体视网膜病变（PVR）发生和发展过程中可促进DNA的合成，引起细胞增生。

Subcellular location and Functional Study of Cell Proliferation-Related Gene CDADC 1 in Human Retinal Pigment Epithelial Cells

Objective： To investigate the expression of cell proliferation-related gene CDADC 1 in human retinal pigment epithelial cells （ARPE19） and its effect on the proliferation of ARPE19. Methods： Fluorescence expressing vector pEGFP-C1-CDADC1 and eu- karyotic expressing vector pcDNA3.1-myc-CDADC 1 were constructed by using gene recombination technique. After confirmed by PCR and DNA sequencing, the plasmids were transfected into ARPE19 cells using liposome-mediated transfecting methods. The subcellular location of GFP-CDADC 1 fusion proteins in ARPE 19 cells was detected. Cell cycle distribution and apoptosis of ARPE 19 cells transfected by pcDNA3.1-myc-CDADC1 was determined by using flow cytometry. Results： Confirmed by PCR and DNA sequencing, the fluorescence expressing vector pEGFP-C 1-CDADC 1 and eukaryotic expressing vector pcDNA3.1-myc-CDADC 1 were successfully constructed. Fusion has low expression in the cytoplasm, while has high expression in the nucleus of ARPE 19 cells. The results of transient transfection of pcDNA3.1-myc-CDADC 1 into ARPE 19 cells indicated that 24th hour after transfection, there was no change of the cells, but 48th hour later, the average percentage of RPE cells which entered the S stage of cell cycle was 19.37 for pcDNA3.1-myc-CDADC1, 10.87 for pcDNA3.1-myc and 3.33 for control. The differences between the three groups were statistically significant （P〈0.01）. Conclusion： CDADCI could promote DNA synthesis and induce cell proliferation in the pathogenesis and development of PVR.