摘要目的: 探讨过表达SOX11(Sex determination region of Y chromosome 11)基因对人视网膜母细胞瘤Y79细胞增殖、凋亡的影响及参与该过程的细胞信号传到途径。 方法: 将用重组pcDNA3.1-SOX11质粒转染的Y79细胞作为实验组,以空载体pc-DNA3.1-mock转染的细胞作为对照组。细胞增殖通过Edu细胞荧光染色法检测,细胞凋亡通过Hoechst 33342细胞染色法检测。PTEN、AKT及p-AKT的表达采用westernblot法检测。 结果: 实验组中SOX11 mRNA的相对表达量显著高于对照组[(354±26.2)% vs (100±1.3)%,P<0.05 ]。实验组细胞在450nm吸光(OD)值低于对照组(2.8±0.3 vs 4.5±0.3,P<0.05)。Edu免疫荧光染色结果证实,实验组细胞增殖比例较对照组低[(12.4±4.3)% vs (32.3±3.3)%,P<0.05]。Hoechst 33342细胞凋亡荧光染色结果表明,实验组细胞凋亡比例较对照组高[(10.3±2.3)% vs (5.3±1.4)%,P<0.05]。Western blot实验结果表明,实验组PTEN蛋白的相对表达量高于对照组(0.6±0.03 vs 0.3±0.03,P<0.05),磷酸化Akt的相对表达量低于对照组(0.4±0.02 vs 0.7±0.02,P<0.05)。 结论: 过表达SOX11基因显著抑制人视网膜母细胞Y79的增殖并促使细胞凋亡。PTEN/Akt信号通路的激活可能参与该过程。
Abstract:Objective: To explore the effects of SOX11 overexpression on proliferation and apoptosis of retinoblastoma of Y79 cells and the underlying mechanism. Methods: Y79 cells were transfected with pcDNA3.1-SOX11 plasmid or pc-DNA3.1-mock. Stable overexpression of SOX11 Y79 cells were collected for the following functional experiments. The effects of SOX11 overexpression on cell proliferation were examed by Edu cell immunofluorescence staining assay. Cell apoptosis were analyzed by Hoechst 33342 staining assay. Akt phosphorylation and PTEN protein expression were detected by Western blot assay. Results: The expression of SOX11 mRNA in SOX11 overexpression group were higher than control group [(354±26.2)% vs (100±1.3)%,P<0.05]. The OD 450nm value of SOX11 overexpression group were lower than control group(2.8±0.3 vs 4.5±0.3,P<0.05). The rate of proliferation cells in SOX11 overexpression group were decreased compared with control group [(12.4±4.3)% vs (32.3±3.3)%,P<0.05] detected by Edu cell immunofluorescence staining assay. The percentage of apoptosis cells in SOX11 overexpression group were higher than control group [(10.3±2.3)% vs (5.3±1.4)%,P<0.05] detected by Hoechst 33342 staining assay. The phosphorylation of AKT [(0.7±0.02)vs(0.4±0.02),P<0.05] was decreased, while PTEN expression [(0.3±0.03)vs(0.6±0.03),P<0.05]increased in SOX11 overexpression group compared with control group. The difference had statistical significance. Conclusion: SOX11 ovrexpression can inhibit Y79 cell proliferation and enhance apoptosis. PTEN/Akt signal pathway may involve in this process.
黄晓寒, 唐少华, 刘爽, 董红. 过表达SOX11基因对人视网膜母细胞瘤Y79细胞增殖凋亡影响的研究[J]. 河北医学, 2018, 24(9): 1409-1412.
HUANG Xiaohan, TANG Shaohua, LIU Shuang, et al. Effects of SOX11 Overexpression on Proliferation and Apoptosis of Retinoblastoma of Y79 Cells. HeBei Med, 2018, 24(9): 1409-1412.
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