视网膜母细胞瘤细胞株SO-RB_(50)中生存素的表达

Expression of survivin in SO-RB_(50) cells

目的研究凋亡抑制因子生存素(survivin)在视网膜母细胞瘤细胞株SO-RB50中的表达。方法收集培养的SO-RB50细胞,通过RT-PCR、Western blot和免疫组织化学法检测SO-RB50细胞中survivin mRNA和蛋白的表达情况。免疫组织化学法检测survivin在正常视网膜组织中的表达。结果SO-RB50细胞中可检测到survivin mRNA和蛋白的表达,免疫组织化学检测显示survivin在SO-RB50细胞核和细胞浆中均有表达。正常视网膜组织中未见survivin表达。结论Survivin参与了视网膜母细胞瘤的发病。

Objective Survivin is thought to be the most powerful apoptosis inhibiting factor, which could suppress apoptosis and promote proliferation of cells. Study has proved that the overexpression of survivin might lead to neoplasm. Present study was to investigate the expression of survivin in human retinoblastoma cell line, SO-RB50. Methods Human retinoblastoma cell line, SO-RB50 was cultured in RPMI-1640 medium containing 10% fetal bovine serum. The expression of survivin mRNA in SO-RB50 was detected by RT-PCR, and expression of survivin protein in SO-RB50 was detected by Western blot and immunohistochemistry. The retina of eyeball from donor was prepared for the detect of survivin protein by immunohistocbemistry as control. Results Three bands at 5 s, 18 s and 28 s were clearly showed by agarose gel electrophoresis after extract of total RNA, and the band with 260 bp was verified to be an amplified product of survivin mRNA. Western blot revealed a expression of survivin protein in SO-RBs0 cell with the relative molecular weight 16 500. Immunohistochemistry determined that the survivin protein distributed in both nucleolus and cytoplasm of SO-RB50. However, no survivin protein was detected in normal retina of donor. Conclusion Survivin plays a role in the pathogenesis of retinoblastoma.