| 牛方霞,李金凤,高哲,等.海参糖胺聚糖联合放疗对人肺腺癌A549细胞增殖抑制及促凋亡机制研究[J].中国海洋药物,2020,39(5):16-22. |
| 海参糖胺聚糖联合放疗对人肺腺癌A549细胞增殖抑制及促凋亡机制研究 |
| The mechanism of proliferation inhibition and apoptosis promotion of Holothurian Glycosaminoglycan combined with radiation on human lung adenocarcinoma A549 |
| 投稿时间：2020-01-13 修订日期：2020-07-29 |
| DOI： |
| 中文关键词: 海参糖胺聚糖 放射 A549细胞 增殖抑制 细胞凋亡 放疗增敏 |
| English Keywords:hGAG radiation A549 cells proliferation inhibition apoptosis radiation sensitization |
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| 中文摘要: |
| 目的 探讨海参糖胺聚糖（hGAG）与放疗联合应用对人肺腺癌A549细胞的增殖抑制作用及促凋亡作用机制。方法 体外培养人肺腺癌A549细胞至对数生长期，分为对照组、hGAG组、放射组、联合组（hGAG联合放射处理）。CCK-8法测定不同处理组细胞的增殖抑制率；Hoechest 33258法对实验细胞进行染色并观察细胞变化；AnnexinV-FITC/PI双染法联合流式细胞术检测凋亡状况；蛋白印迹法（Western blot）测定各个组细胞survivin、Bax、Bcl-2及caspase-3四种凋亡蛋白的相对表达量。结果（1）CCK-8法显示，24h、48h和72h各时间段hGAG组、放射组、联合组细胞的增殖抑制率均高于对照组，差异均有统计学意义（P<0.05）；24h、48h和72h各时间段联合组细胞的增殖抑制率亦均高于放射组，差异均有统计学意义（P<0.05）。（2）Hoechest 33258染色示：hGAG组、放射组及联合组在镜下均可见凋亡细胞呈现核碎裂、核固缩现象，细胞呈现为染色浓密的状态，联合组较放射组更为明显；（3）流式细胞仪检测显示:三个实验组的细胞总凋亡率较对照组高（P<0.05），以及联合组的细胞总凋亡率比放射组高（P<0.05）；（4）Western blot示：三个实验组与对照组比较，以及联合组与放射组相比，蛋白Bax、caspase-3的表达水平升高，蛋白Bcl-2、survivin的表达水平降低（均P<0.05）。结论 hGAG可有效抑制A549肺癌细胞的增殖并促进其凋亡，并且可以增强A549细胞系对射线的敏感性，推测其作用机制可能与caspase-3蛋白和Bax蛋白表达上调，以及survivin蛋白、Bcl-2蛋白的表达下调有关。 |
| English Summary: |
| Objective: To investigate the proliferation inhibition and apoptosis promotion of Holothurian Glycosaminoglycan（hGAG） in combination with radiation on pulmonary adenocarcinoma cell line A549 and to study the mechanism of these effects. Methods: The human lung adenocarcinoma A549 cells were cultured to logarithmic growth phase and divided into four groups: control group, hGAG group, radiation group, combination group (hGAG combined radiation). CCK-8 method was used to detect the inhibition rate of cell proliferation in each group. Hoechest 33258 method was used to stain the experimental cells and observe the morphological changes of cell apoptosis. Annexin V-FITC/PI double-staining combined with flow cytometry was used to detect cell apoptosis. Western blot was used to detect the relative expression of four apoptotic proteins (Bcl-2, Bax, survivin and caspase-3). Results: CCK-8 experimental results showed that the proliferation inhibition rate of hGAG group, the radiation group and the combination group were all higher than that of the control group after treatment for 24 h,24h and 48h (all P < 0.05). The proliferation inhibition rates of the combination group were higher than those of the radiation group at each time period (all P < 0.05). Hoechest 33258 staining experiment showed that dense apoptotic cells, nuclear fragmentation and nuclear pyknosis could be observed in hGAG group, radiation group and combination group. The most obvious manifestation was in the combination group. Annexin V-FITC/PI double-staining combined with flow cytometry showed that the total apoptotic rate of the three experimental groups was significantly higher than that of the control group and the total apoptotic rate of the combination group was significantly higher than that of the radiation group (all P < 0.05). Furthermore, western blot results showed that compared with the control group, the expression of protein Bax and caspase-3 were increased and the expression of protein Bcl-2 and survivin were decreased in the three experimental groups. The expression of protein Bax and caspase-3 were increased and the expression of protein Bcl-2 and survivin were decreased in the combination groups compared with the radiation group (all P < 0.05). Conclusion: hGAG can inhibit the proliferation and promote apoptosis the of A549 cells and enhance the radio sensitivity of A549 cells. Its mechanism may be related to up-regulation of Bax and caspase-3 protein expression and down-regulation of Bcl-2 and survivin protein expression. |
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