HXO-RB44细胞上清液对ARPE-19细胞内VEGF表达的影响
摘要
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 supernatant 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 acre cultured in MEM medium containing 10% bovine serum. 100 μL of HXO-RB44 supernatant 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 supernatant fluid. Immunocytochemistry was used to evaluate the expression of VEGE in 4RPE-19 cells based on the fluorescence intensity. Results The expression of VRGF mRNA was significantly different among various time points alter 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-l9 cells was obviously increased (P＜0.01). Expression of VEGE protein was significantly increased in ARPE19 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 VEGF mRNA in the d3namicsl change to HXO-RB44 supernatant fluid (P＜0.05). The green fluorescence intensity ass gradually higher with the time prolong of HXO-RB44 supernatant fluid action. Conclusion Supernatant fluid of HXO-RB cells can enhance the expression of EGF in ARPE-19 cells.