Wang Li-Lun, Feng Yan-Qin, Cheng Yu-Hong
Department of Ophthalmology, Affiliated Hospital of Yan'an University, Yan'an 716000, Shaanxi Province, China.
Department of Ophthalmology, the First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710061, Shaanxi Province, China.
Int J Ophthalmol. 2017 Jan 18;10(1):30-34. doi: 10.18240/ijo.2017.01.05. eCollection 2017.
To investigate the effect of high mobility group protein box-1 (HMGB1) siRNA on proliferation and apoptosis of retinoblastoma (Rb) cells.
The expression of HMGB1 in Rb cells were detected by real-time polymerase chain reaction (RT-PCR) and Western blot. Chemically synthesized HMGB1 siRNA was transfected into Y79 cells. The inhibitory rate was also examined by RT-PCR and Western blot. After HMGB1 siRNA transfection, the cell proliferation was analyzed by MTT, and cell apoptosis was detected by Caspase-3 active detection kit. Cell cycle distribution and apoptosis were detected by flow cytometry.
The expression of HMGB1 significantly elevated in Rb cells (<0.01). After transfected by siRNA, the HMGB1 protein level of Y79 cells was significantly reduced (<0.01). After siRNA interference HMGB1, the proportion of proliferating cells reduced, and the proportion of quiescent cells increased (<0.05). In addition, apoptosis rate of Y79 cells increased from 2.03% to 9.10% after interfering with HMGB1 siRNA (<0.05).
Specific HMGB1 siRNA can inhibit the expression of HMGB1. The effect may be attributed to inhibit the proliferation and promote cell apoptosis.
探讨高迁移率族蛋白盒1（HMGB1）小干扰RNA（siRNA）对视网膜母细胞瘤（Rb）细胞增殖和凋亡的影响。
采用实时聚合酶链反应（RT-PCR）和蛋白质印迹法检测Rb细胞中HMGB1的表达。将化学合成的HMGB1 siRNA转染至Y79细胞。同时采用RT-PCR和蛋白质印迹法检测干扰效率。转染HMGB1 siRNA后，采用噻唑蓝（MTT）法分析细胞增殖情况，用Caspase-3活性检测试剂盒检测细胞凋亡情况。通过流式细胞术检测细胞周期分布和凋亡情况。
Rb细胞中HMGB1的表达显著升高（<0.01）。转染siRNA后，Y79细胞中HMGB1蛋白水平显著降低（<0.01）。干扰HMGB1后，增殖细胞比例降低，静止细胞比例增加（<0.05）。此外，干扰HMGB1 siRNA后，Y79细胞凋亡率从2.03%升至9.10%（<0.05）。
特异性HMGB1 siRNA可抑制HMGB1表达，其作用机制可能是抑制细胞增殖并促进细胞凋亡。
