| 康亚辉,葛宁,洪福,等.穿心莲内酯对食管癌细胞的放射增敏作用[J].安徽医药,2019,23(12):2517-2521. |
| 穿心莲内酯对食管癌细胞的放射增敏作用 |
| Radiosensitivity of Andrographolide on esophageal cancer cell |
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| DOI：10.3969/j.issn.1009?6469.2019.12.046 |
| 中文关键词: 食管肿瘤 辐射增敏药 穿心莲内酯 辐射耐受性 bcl?2相关 X蛋白质 NF?κB 半胱氨酸天冬氨酸蛋白酶 3 组蛋白类 |
| 英文关键词: Esophageal neoplasms Radiation?sensitizing agents Andrographolide Radiation tolerance Bcl?2?associated X protein NF?kappa B Caspase 3 Histones |
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| 中文摘要: |
| 目的研究穿心莲内酯对食管癌细胞的放射增敏作用。方法培养食管癌 ECA?109细胞，通过四甲基偶氮唑盐微量酶反应比色法(MTT法)检测穿心莲内酯对细胞生长的抑制；克隆集落形成实验观察穿心莲内酯的放射增敏作用；通过免疫荧光实验观察促凋亡蛋白 Bax的表达情况；流式细胞仪检测细胞的凋亡和周期情况；蛋白质印迹法(Western Blot)检测细胞内核因子 κB(NF?κB)半胱氨酸天冬氨酸蛋白酶 3(caspase3)Bax，B淋巴细胞瘤 ?2基因(Bcl?2)的表达；磷酸化组蛋白 H2AX(γ? H2AX)焦点形成检，测 DNA分钟损伤情况。细胞划痕实验，检测细胞增殖、迁移的情况。结果穿心莲内酯抑制食管癌 ECA? 109细胞的生长，并且有明显的时间和剂量依赖性；通过单击靶模型拟合曲线，可见低浓度穿心莲内酯预处理 6h，相对比单纯照射组可以增加 ECA?109细胞的放射敏感性(t＝4.59，P＝0.02)放射增敏比为 1.28；与单纯照射组(26.5±2.2)%相比，加入穿心莲内酯联合 X射线照射组中的细胞凋亡率(32.5±0.9)%明显增加，(P＜0.05)；穿心莲内酯作用于食管癌细胞，可以使凋亡相关蛋白 caspase3，Bax的表达升高，并且呈明显的量效关系， NF?κB表达趋势与 Bcl?2一致；相对于单纯照射组，穿心莲内酯与 X射线联合作用，能够增加双链 DNA断裂(DSB)数目；药物联合放射可以明显影响食管癌细胞的增殖迁移能力。结论穿心莲内酯能够提高食管癌细胞的放射敏感性，机制可能与其增加食管癌细胞的凋亡率和调节细胞内的 NF?κB的表达有关。 |
| 英文摘要: |
| Objective To explore the radiosensitivity of Andrographolide on esophageal cancer.Methods The inhibition effects of Andrographolide were measured by MTT assay.Clonogenic survival assay was used to evaluate the effect of Andrographolide on ra? diosensitivity of esophageal cancer cell.Immunofluorescence was employed to examine the expression of Bax.The change of cellapoptosis was assayed by flow cytometry.The expression of NF?κB/caspase3/Bax/Bcl?2 was measured by Western Blot.DNA damagewas detected by γ?H2AX foci counting.Results With a clear dose and time effect，Andrographolide could inhibit the proliferation of esophageal cell line ECA?109.Clonogenic survival assay showed that Andrographolide could markedly enhance the radiosensitivity(t＝4.59，P＝0.02)and the sensitizing enhancement ratio(SER)was 1.28.Compared with the irradiation group(26.5±2.2)%，the apoptosis rate in the Andrographolide combined with X?ray irradiation group(32.5±0.9)% was significantly increased(P＜0.05). Andrographolide caused a dose?dependent increase in caspase3/Bax protein expressions，in contrast Bcl?2/NF?κB；Compared with control，the apoptosis in ECA?109 treated with Andrographolide was increased significantly；Compared with radiation alone group， Andrographolide combined with the X?ray can increase the number of DNA double chain brea(DSB).Conclusion Androgra? pholide can increase the radiosensitivity of esophageal cell line ECA?109，which may be associated with reducing the level of NF? κB and inducing apoptosis of esophageal cancer cells. |
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