AG490对人视网膜母细胞瘤HXO-RB44细胞系中JAK2/STAT3信号通路的影响
[Effect of AG490 on JAK2/STAT3 signaling pathway in human retinoblastoma HXO-RB44 cell lines].
作者信息
Xu Bei, Chen Xiang, Tan Jia, Xu Xueliang
机构信息
Department of Ophthalmology, Xiangya Hospital, Central South University, Changsha 410008, China.
Department of Dermatology, Xiangya Hospital, Central South University, Changsha 410008, China.
出版信息
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2018 Oct 28;43(10):1061-1067. doi: 10.11817/j.issn.1672-7347.2018.10.004.
To investigate the role of Janus kinase (JAK) inhibitor AG490 in the anti-proliferation and cell cycle in human retinoblastoma HXO-RB44 cell lines in vitro, and to explore its effect on the expression of JAK2/signal transducer and activator of transcription 3 (STAT3). Methods: Cells were divided into an experiment group and a control group, and the experiment group was further divided into 6 sub-groups according to different AG490 concentrations (6.25, 12.50, 25.00, 50.00 or 100.00 μmol/L). Cell proliferation in the different groups was analyzed by cell vitality determination. Cell cycle distribution and apoptosis rate were examined by flow cytometry. The protein levels of STAT3, p-STAT3 and vascular endothelial growth factor (VEGF) were detected by Western blot. Results: After 48 h treatment with AG490, the viability of HXO-RB44 cells was reduced in a concentration-dependent manner. Compared with the control group, there was no significant difference in the experiment groups except the 6.25 μmol/L group (all P>0.05). The apoptosis rates in the experiment groups were significantly increased with increase in concentration of AG490 compared with that in the control group (all P<0.05). The cell ratio in the G1 phase in 50 or 100 μmol/L group was increased, whereas the cell ratio in the S phase was decreased. Western blot results showed that the expressions of STAT3 and p-STAT3 in the experiment groups were dramatically reduced with the increase in concentration of AG490 compared with that in the control group (all P<0.05). VEGF expression didn't obviously change in the experiment groups with AG490 concentration less than 12.5 μmol/L compared with that in the control group (both P>0.05), but there were significant differences in the other experiment groups (all P<0.05). Conclusion: JAK inhibitor AG490 can inhibit proliferation and promote apoptosis of the retinoblastoma HXO-RB44 cells through down-regulation of JAK2/STAT3 signaling pathway.
为研究Janus激酶（JAK）抑制剂AG490在体外对人视网膜母细胞瘤HXO-RB44细胞系增殖及细胞周期的作用，并探讨其对JAK2/信号转导及转录激活因子3（STAT3）表达的影响。方法：将细胞分为实验组和对照组，实验组再根据不同AG490浓度（6.25、12.50、25.00、50.00或100.00 μmol/L）分为6个亚组。通过细胞活力测定分析不同组的细胞增殖情况。采用流式细胞术检测细胞周期分布及凋亡率。通过蛋白质印迹法检测STAT3、p-STAT3和血管内皮生长因子（VEGF）的蛋白水平。结果：AG490处理48 h后，HXO-RB44细胞活力呈浓度依赖性降低。与对照组相比，除6.25 μmol/L组外，实验组差异均无统计学意义（均P>0.05）。与对照组相比，实验组凋亡率随AG490浓度增加而显著升高（均P<0.05）。50或100 μmol/L组G1期细胞比例增加，S期细胞比例降低。蛋白质印迹结果显示，与对照组相比，实验组中STAT3和p-STAT3的表达随AG490浓度增加而显著降低（均P<0.05）。AG490浓度小于12.5 μmol/L的实验组与对照组相比，VEGF表达无明显变化（均P>0.05），但其他实验组差异有统计学意义（均P<0.05）。结论：JAK抑制剂AG490可通过下调JAK2/STAT3信号通路抑制视网膜母细胞瘤HXO-RB细胞的增殖并促进其凋亡。
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