附子提取物对肺癌A549细胞增殖抑制及放射增敏作用的研究

doi:10.3969/j.issn.1004-616x.2022.03.010

摘要/Abstract

摘要： 目的: 观察附子提取物对人肺癌A549细胞增殖及放射敏感性的影响,并探讨其可能的机制。方法: 体外培养肺癌A549细胞,采用CCK-8法检测不同浓度附子提取物作用后细胞增殖情况,并选择IC₂₀(30μg/mL)作为后续实验浓度。集落形成实验检测A549细胞的存活分数,单机多靶模型拟合细胞存活曲线计算平均致死剂量(D₀)及放射增敏比(SER)。将细胞分为对照组、附子组、X射线照射组和附子处理后X射线照射组。对照组给予培养基,附子组给予30μg/mL附子提取物处理24 h,X射线照射组采用10 Gy的X射线照射,附子处理后X射线照射组先采用30μg/mL附子提取物处理24 h后,再给予10 Gy的X射线照射。流式细胞术检测细胞凋亡率,Western blot法检测细胞中Bax、Bcl-2蛋白表达水平。结果: CCK-8法检测结果显示,小于20μg/mL的附子提取物对A549细胞存活率有增强作用,但随着附子提取物浓度到达30μg/mL,开始对人肺癌A549细胞生长具有抑制作用,且与附子提取物浓度相关。集落形成实验结果提示附子处理后X射线照射组存活曲线起始处肩部稍窄,表明联用附子提取物后,引起细胞死亡所需的放疗剂量逐渐减小,X射线照射组和附子处理后X射线照射组D₀值分别为1.83和1.48 Gy,SER为1.24。流式细胞术检测结果提示附子处理后X射线照射组细胞凋亡率显著增加,高于附子组和X线照射组(P<0.05)。Western blot检测结果显示,附子组与X射线照射组较对照组细胞Bax蛋白表达水平升高,Bcl-2蛋白表达水平下降。附子处理后X射线照射组较对照组细胞Bax、Bcl-2蛋白含量变化更加显著(P<0.01)。结论: 30μg/mL的附子提取物对人肺癌A549细胞具有增殖抑制作用和放射增敏作用,其机制可能与诱导细胞凋亡有关。

关键词: 肺癌, 附子提取物, 中药治疗, 放射治疗, 放疗增敏, 细胞凋亡

Abstract: OBJECTIVE: To study effects of Fuzi (aconite) extracts on proliferation and radiosensitization of human lung cancer A549 cells. METHODS: The cells were cultured and were divided into 4 groups:control,Fuzi extract,X-ray-irradiated,and Fuzi plus X-ray-irradiated groups. The control group was given culture medium,the Fuzi extract group was treated with IC₂₀ 30 μg/mL of Fuzi extract,the radiation group was given X-ray-irradiated,and the Fuzi plus X-ray-irradiated group were first treated with the Fuzi extract and then X-ray. After treatments,cell proliferation was detected by CCK-8 assay and colony formation assay was used to measure cell survival. Single-hit multi-target model was used to fit the survival curve and to calculate the sensitive enhancement ratio (SER). Apoptosis rates were detected by flow cytometry,and protein expression levels of Bax and Bcl-2 were detected by Western blot. RESULTS: Fuzi extract concentrations(<20 μg/mL) enhanced viability of A549 cells but increasing concentrations (>30 μg/mL) inhibited growth of the cells in a dose-dependent manner. The shoulder at the beginning of the survival curve in the Fuzi plus X-ray-irradiated group was slightly narrower, and the D₀values in the X-ray-irradiated and Fuzi plus X-ray-irradiated group were 1.83 Gy,1.48 Gy,and the SER was 1.24,respectively. The apoptosis rates were significantly increased in the Fuzi plus X-ray-irradiated group and were higher than that in both the Fuzi and X-ray irradiation groups (P<0.05). The results of Western blot show that protein expressions of Bax were increased but were decreased for Bcl-2 in the Fuzi extract versus the X-ray-irradiated groups. Bax and Bcl-2 protein levels were changed more significantly in the Fuzi plus X-ray-irradiated than the other groups (P<0.01). CONCLUSION: Fuzi extract at 30 μg/mL inhibited proliferation and enhanced radiosensitizing effects on human lung cancer A549 cells,and the mechanism may be related to the induction of apoptosis.

Key words: lung neoplasms, Fuzi extraction, Chinese medicine treatment, radiotherapy, radiosentization, cell apoptosis

中图分类号:

R730.1

张琪, 刘宇峻, 庄喜兵, 乔田奎. 附子提取物对肺癌A549细胞增殖抑制及放射增敏作用的研究[J]. 癌变·畸变·突变, 2022, 34(3): 227-232.

ZHANG Qi, LIU Yujun, ZHUANG Xibing, QIAO Tiankui. Fuzi extracts on proliferation and radiosensitization of lung cancer A549 cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2022, 34(3): 227-232.

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