褪黑素对人非小细胞肺癌H1299细胞辐射敏感性的影响

doi:10.13481/j.1671-587x.20180314

摘要/Abstract

摘要： 目的：检测褪黑素（MLT）联合放疗对人非小细胞肺癌（NSCLC） H1299细胞增殖和凋亡的影响，阐明MLT在调节H1299细胞辐射敏感性中的作用。方法：将H1299细胞分为对照组、MLT组、照射组及照射+MLT组，对照组H1299细胞不作任何处理，MLT组H1299细胞给予不同浓度（100和500 μmol·L^-1） MLT，照射组H1299细胞采用¹³⁷Cs γ射线进行一次性6 Gy照射，照射+MLT组H1299细胞给予100和500μmol·L^-1 MLT加6 Gy¹³⁷Cs γ射线。克隆形成实验检测各组细胞克隆形成数，流式细胞术检测照射后24和48 h各组细胞凋亡率，蛋白免疫印迹法检测各组细胞中核转录因子κB（NF-κB）蛋白的磷酸化水平。结果：克隆形成实验，与照射组比较，照射+MLT组H1299细胞克隆形成数明显减少（t=7.234，P<0.01）。流式细胞术，照射后24 h，与对照组比较，照射组和MLT组H1299细胞凋亡率明显升高（t=5.020，t=13.525，P<0.01），与对照组比较，照射+MLT组H1299细胞凋亡率升高（t=12.884，P<0.01）；照射后48 h，照射+MLT组H1299细胞凋亡率与照射组比较差异无统计学意义（t=0.394，P>0.05）。蛋白免疫印迹法，照射+MLT组H1299细胞中NF-κB蛋白磷酸化水平低于照射组。结论：MLT联合放疗对人NSCLC H1299细胞有明显的抑瘤效应，提高了NSCLCH1299细胞的辐射敏感性。

关键词: 褪黑素, 细胞增殖, 细胞凋亡, 癌, 非小细胞肺, 核转录因子κB, 辐射敏感性

Abstract: Objective: To detect the effects of melatonin (MLT) combined with radiotherapy on the proliferation and apoptosis of non-small cell lung cancer (NSCLC) H1299 cells,and to explore the role of MLT in regulating the radiosensitivity of H1299 cells. Methods: The H1299 cells were divided into control group,MLT group,irradiation(IR) group and IR+MLT group. The H1299 cells in control group didn't receive any treatment, the H1299 cells in MLT group were treated with different concentrations of MLT (100 and 500 μmol·L^-1),the H1299 cells in IR group were exposed to 6 Gy ¹³⁷Cs γ-ray,and the H1299 cells in IR+MLT group were treated with different concentrations of MLT (100 and 500 μmol·L^-1) and exposed to 6 Gy ¹³⁷Cs γ-ray. Colony formation assay was used to detect the clone numbers of H1299 cells in various groups; Flow cytometry was applied to detect the apoptotic rates of H1299 cells at 24 and 48 h after irradiation; Western blotting method was performed to detect the phosphorylation levels of nuclear factor kappa B (NF-κB) protein in the H1299 cells in various groups. Results: The colony formation assay results showed that compared with IR group,the clone number of H1299 cells in IR+MLT group was significantly decreased(t=7.234,P<0.01).The flow cytometry results showed that the apoptotic rates of H1299 cells in IR group and MLT group at 24 h after irradiation were higher than that in control group (t=5.020,t=13.525,P<0.01); the apoptotic rate of H1299 cells in IR+MLT group at 24 h after irradiation was higher than that in IR group (t=12.884,P<0.01).There was no significant difference in the apoptotic rate of the H1299 cells between IR+MLT group and IR group at 48 h after irradiation(t=0.394,P>0.05).The Western blotting results showed that the phosphorylation level of NF-κB protein in the H1299 cells in IR+MLT group was lower than that in IR group. Conclusion: MLT combined with radiotherapy has obviously inhibitory effect on the human H1299 cells and can increase the radiosensitivity of H1299 cells.

Key words: melatonin, nuclear factor kappa B, cancer,non-small cell lung, proliferation, apoptosis, radiosensitivity

中图分类号:

Q691

杨文延, 刘强, 孙志娟, 杜利清, 徐畅, 王彦, 柳杨, 王芹. 褪黑素对人非小细胞肺癌H1299细胞辐射敏感性的影响[J]. 吉林大学学报(医学版), 2018, 44(03): 532-536.

YANG Wenyan, LIU Qiang, SUN Zhijuan, DU Liqing, XU Chang, WANG Yan, LIU Yang, WANG Qin. Effect of melatonin on radiosensitivity of non-small cell lung cancer H1299 cells[J]. Journal of Jilin University Medicine Edition, 2018, 44(03): 532-536.

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