P21 抑制剂 UC2288 诱导鼻咽癌放射抗拒细胞 CNE-2R 凋亡的研究
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1.广西医科大学附属肿瘤医院,南宁 530021; 2.广西区域高发肿瘤早期防治重点实验室,南宁 530021; 3.广西医科大学附属武鸣医院,南宁 530021

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Apoptosis of nasopharyngeal carcinoma radioresistant cell line CNE-2R induced by P21 inhibitor UC2288
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1.Guangxi Medical University Cancer Hospital, Nanning 530021, China. 2. Key Laboratory of Early Prevention and Treatment for Regional High Frequency Tumor (Guangxi), Ministry of Education, Nanning 530021. 3. Wuming Hospital of Guangxi Medical University, Nanning 530021

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    摘要:

    目的 探讨 P21 抑制剂 UC2288 对鼻咽癌放射抗拒细胞 CNE-2R 增殖和凋亡的影响及可能的作用机制。 方法 运用 CCK-8 实验检测 UC2288 对细胞活力的影响;利用克隆形成实验检测细胞克隆形成能力的情况; 显微镜拍照法和 Hoechst 33342 染色实验观察细胞形态的变化;Annexin V-APC/ 7-AAD 双染法测定细胞凋亡;蛋白免疫印迹法(Western blot)检测 Bax、Cleaved-caspase 3、Caspase 3、Bcl-2、Survivin、γ-H2AX、P21、聚腺苷酸二磷酸核糖 聚合酶(PARP)蛋白表达情况。 结果 CCK-8 实验结果显示,UC2288 显著降低 CNE-2R 细胞的活力和增殖,呈剂量依赖性和时间依赖性;UC2288 抑制 CNE-2R 细胞的克隆形成能力;形态学观察结果显示,UC2288 处理后,细胞呈圆形萎缩,细胞核固缩,体积变小;Annexin V-APC/ 7-AAD 实验结果显示,UC2288 呈剂量依赖性诱导 CNE-2R 细胞凋亡;UC2288 作用细胞后,Bax、Cleaved-caspase 3、Caspase 3 和 γ-H2AX 蛋白表达水平增加,Bcl-2、Survivin、P21 和 PARP 蛋白表达量减少。 结论 UC2288 显著抑制 CNE-2R 细胞增殖,引起 DNA 损伤,诱导凋亡,其机制可能与 PARP 表达水平降低有关。

    Abstract:

    Objective To assess the effects of P21 inhibitor UC2288 on proliferation and apoptosis of nasopharyngeal carcinoma radioresistant cell line CNE-2R and the possible mechanisms. Methods The effect of UC2288 on cell viability was examined by a CCK-8 assay. The colony formation ability was investigated by a colony formation assay. Changes of cell morphology were observed by microscopy and Hoechst 33342 staining. An annexin V-APC/ 7-AAD assay was used to measure apoptosis. The protein levels of Bax, Cleaved-caspase 3, Caspase 3, Bcl-2, Survivin, γ-H2AX, P21, and PARP were measured by Western blot. Results The CCK-8 assay showed that UC2288 significantly reduced the viability and proliferation of CNE-2R cells in dose- and time-dependent manners. UC2288 also inhibited the colony formation of CNE-2R cells. Cells became round and atrophic, the nucleus shrunk, and the cell volume became smaller after treatment with UC2288. Annexin V-APC/ 7-AAD assays demonstrated that UC2288 induced apoptosis of CNE-2R cells in a dose-dependent manner. After treatment with UC2288, the expression levels of Bax, Cleaved-caspase 3, Caspase 3, and γ- H2AX increased accompanied by decreases of Bcl-2, Survivin, P21, and PARP. Conclusions UC2288 significantly inhibits the proliferation of CNE-2R cells, causes DNA damage, and induces apoptosis through possibly reducing the expression of PARP.

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梁仁拔,李欣晓,朱小东. P21 抑制剂 UC2288 诱导鼻咽癌放射抗拒细胞 CNE-2R 凋亡的研究[J].中国比较医学杂志,2020,30(8):16~22.

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  • 收稿日期:2020-01-09
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  • 在线发布日期: 2020-09-02
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