摘要
目的探讨HXO-RB44细胞上清液对人视网膜色素上皮(RPE)细胞系ARPE-19细胞内血管内皮生长因子(VEGF)表达的影响及其相关机制。方法分别利用含有10%胎牛血清的RPMI-1640培养基及MEM培养基对HXO-RB44细胞及ARPE-19细胞进行培养,在ARPE-19细胞内加入100μL HXO-RB44细胞上清液,分别干预0、4、8、12、24 h,利用RT-PCR、免疫荧光细胞化学、Western blot等方法,检测不同浓度的MLT干预后24 h ARPE-19细胞内VEGF mRNA及蛋白的表达情况。结果HXO-RB44细胞上清液加入后的不同时间点,VEGF mRNA的表达差异有统计学意义(F=195.072,P=0.000)。随着HXO-RB44细胞上清液作用时间的延长,ARPE-19细胞内VEGF mRNA表达逐渐增高(P<0.01)。HXO-RB44细胞上清液加入后4、8、12、24 h,ARPE-19细胞内VEGF蛋白表达显著增加(F=160.687,P=0.000),其表达与ARPE-19细胞内VEGF mRNA表达方式相同(P<0.05)。随着HXO-RB44细胞上清液作用时间的延长,ARPE-19细胞内绿荧光蛋白强度逐渐增加。结论HXO-RB44细胞上清液能促进ARPE-19细胞内VEGF的表达。
Objective VEGF is a promoting angiogenesis factor in retinoblastoma. It is associated with the differentiation and extension of tumor. HXO-RB44 supernatant fluid is a RB cell supernatant fluid. Present study was to explore the effect of RB cell supernalanl fluid on expression of VEGF in ARPE-19 cells, a type of retinal pigment epithelial cell, and its related mechanism. Methods HXO-RB44 cells were cultured in RPMI-1640 medium containing 10% bovine serum, and ARPE-19 cells were cultured in MEM medium containing 10% bovine serum. 100 μL of HXO-RB44 supernatanl fluid was added in ARPE- 19 cells medium. The expression of VEGF mRNA in ARPE-19 cells was monitored by RT-PCR, and the expression of VEGF protein was detected using Western-blot analysis in 0, 4, 8, 12, 24 hours alter effect of HXO-RB44 supernatanl fluid. lmmunocytoehemistry was used to evaluate the expression of VEGF in ARPE-19 cells based on the fluorescence intensity. Results The expression of VEGF mRNA was significantly different among various time points after addition of HXO-RB44 supernatant fluid（ F = 195. 072,P = 0. 000）. With the prolong of action time of HXO-RB44 supernatant fluid, the expression of mRNA in ARPE-19 cells was obviously increased（ P 〈 0.01 ）. Expression of VEGF protein was significantly iucreased in ARPE- 19 cells incubated with supernatant fluid of HXO-RB44 cells after 4,8,12,24 hours, showing a considerable increase among different groups （F =160. 687,P =0. 000） and followed the same pattern to VEGE mRNA in the dynamical cbange to HXO-RB44 supernatant fluid （ P 〈 0.05 ）. The green fluorescence intensity was gradually higher with the time prolong of HXO-RB44 supernatant fluid action. Conclusion Supernatant fluid of HXO-RB44 cells can enhance the expression of VEGF in ARPE-19 cells.
出处
《眼科研究》
CSCD
北大核心
2009年第10期878-882,共5页
Chinese Ophthalmic Research
