二氢杨梅素预干预对人肺癌A549细胞放射治疗的敏感性作用

孙大永 毕红东 梁鸿鹄 左彦珍* 王爱民 焦海涛

doi:10.16098/j.issn.0529-1356.2015.06.012

解剖学报 ›› 2015, Vol. 46 ›› Issue (6) : 797-801. DOI: 10.16098/j.issn.0529-1356.2015.06.012

肿瘤生物学

二氢杨梅素预干预对人肺癌A549细胞放射治疗的敏感性作用

孙大永¹毕红东² 梁鸿鹄¹左彦珍^2* 王爱民³焦海涛^4

作者信息 +

Radiosensitivity on A549 cells by dihydromyricetin pre-intervention in vitro

SUN Da-yong¹BI Hong-dong² LIANG Hong-hu¹ ZUO Yan-zhen^2*WANG Ai-min³ JIAO Hai-tao⁴

Author information +

文章历史 +

摘要

目的探索二氢杨梅素（DMY）对人肺腺癌A549细胞放射治疗敏感性的调节作用，及新的肺癌放疗增敏剂。方法采用MTT法和集落形成实验分别寻找二氢杨梅素对A549细胞的短期和长期20%抑制浓度（IC₂₀）作为放疗增敏浓度；流式细胞术周期实验检测二氢杨梅素（IC₂₀）作用不同时间对A549细胞周期时相的改变；集落形成实验检测二氢杨梅素（IC₂₀）预干预24h后再照射对肺癌A549细胞放疗敏感性的影响。结果 MTT和集落形成实验均显示，DMY对A549细胞以浓度依赖形式存在短期和长期的抑制作用，MTT实验时IC₂₀ 为 40mg/L，集落形成实验时IC₂₀为7mg/L；流式细胞术周期实验结果显示，二氢杨梅素作用24h时G2/M期比率增高明显（P<0.05）；集落增敏实验结果显示，DMY预处理24h后对A549细胞的放射治疗并未见到显著增敏作用(P>0.05)。结论二氢杨梅素体外存在抑制A549细胞增殖的作用，其效应与药物浓度和细胞初始接种密度有关；二氢杨梅素作用24h可以使A549细胞G2/M期比率明显增加，但是单纯通过调整放疗前细胞周期时相并未见到明显的放疗增敏作用。

Abstract

Objectiv To study the radiosensitivity of dihydromyricetin (DMY) on human non-small cell lung cancer A549 cells, and to explore a new drug for lung cancer radiotherapy sensitization.Methods MTT assay and colony formation assay were used to determined the IC₂₀ of dihydromyricetin at different density A549 cells. Flow cytometry (FCM) was used to detect the change of A549 cell cycle phase impacted by DMY of IC₂₀during different time (6, 12, 24, 48hours). Colony formation assay was used to determine the radiosensitivity on A549 cells by dihydromyricetin pre-intervention in vitro. Results The proliferation of A549 cells was inhibited in a dose-dependent manner, MTT assay showed the IC20 was 40mg/L and colony formation assay showed the IC₂₀was 7mg/L. FCM cycle experiment showed that the rate of G2/M phase ratio obviously increased when A549 cells was incubated 24 hours with DMY；A significant sensibilization leading by the effect of radiotherapy sensibilization on A549 cells after DMY pre-intervention 24 hours was not detected by clone formation sensibilization assay.Conclusion The proliferation of A549 cells was inhibited in a dose-dependent manner of dihydromyricetin, and the effect was influenced by cell density. Although the rate of G₂/M phase ratio obviously increased when A549 cells were incubated 24 hours with DMY, there was no significant sensibilization on A549 cells after DMY pre-intervention 24 hours. A further study should be made.

导出引用

孙大永毕红东梁鸿鹄左彦珍* 王爱民焦海涛. 二氢杨梅素预干预对人肺癌A549细胞放射治疗的敏感性作用[J]. 解剖学报. 2015, 46(6): 797-801 https://doi.org/10.16098/j.issn.0529-1356.2015.06.012

SUN Da-yong BI Hong-dong LIANG Hong-hu ZUO Yan-zhen*WANG Ai-min JIAO Hai-tao. Radiosensitivity on A549 cells by dihydromyricetin pre-intervention in vitro[J]. Acta Anatomica Sinica. 2015, 46(6): 797-801 https://doi.org/10.16098/j.issn.0529-1356.2015.06.012

参考文献

［1］Zhao WY, Sun XCh. Progression of tumor radiosensitization［J］. Chinese Clinical Oncology, 2012, (7): 655-659. (in Chinese)
赵维勇, 孙新臣. 放射治疗增敏的研究现状［J］. 临床肿瘤学杂志, 2012, (7): 655-659.
［2］Cen JH. Study on apoptosis-inducing mechanism of dihydromyricetin on human liver cancer HepG2［J］. The Journal of Practical Medicine, 2014, (11): 1712-1715. (in Chinese)岑钧华. 二氢杨梅素诱导肝癌细胞株HepG2凋亡途径的探讨［J］. 实用医学杂志, 2014, (11): 1712-1715.
［3］Hou XL, Wang WQ, Shi ChY, el al. Research progress in pharmacological effects of dihydromyricelin［J］. Chinese Traditional and Herbal Drugs, 2015, 46(4): 603-609. (in Chinese)
侯小龙, 王文清, 施春阳,等. 二氢杨梅素药理作用研究进展［J］. 中草药, 2015, 46(4): 603-609.
［4］Yu CC, Hung SK, Liao HF, et al. RAD001 enhances the radiosensitivity of SCC4 oral cancer cells by inducing cell cycle arrest at the G2/M checkpoint［J］. Anticancer Res, 2014,34(6): 2927-2935.
［5］Zhou FZh, Zhang XY, Guo Y. Research progress on antitumor effect and mechanisms of dihydromyricetin［J］. Chinese Archives of Traditional Chinese Medicine, 2010, 28(9):1914-1916. (in Chinese)
周防震, 张晓元, 郭勇. 二氢杨梅素抗肿瘤活性及其机制的研究进展［J］. 中华中医药学刊, 2010,28(9):1914-1916.
［6］Zhou FZh, Zhang XY, Sun FY, et al. Anti proliferation effect of dihydromyricetin on MDA-MB-231 cell in vitro［J］. Cancer Research on Prevention and Treatment, 2012, 39(1): 95-97. (in Chinese)
周防震, 张晓元, 孙奋勇, 等. 二氢杨梅素对人乳腺癌细胞MDA-MB-231的体外抗增殖作用［J］. 肿瘤防治研究, 2012, 39(1): 95-97.
［7］Franken NA, Rodermond HM, Stap J, et al. Clonogenic assay of cells in vitro［J］. Nat Protoc, 2006,1(5):2315-2319.
［8］Yuan ZhY, Liu XB, Zhuang HQ, et al. Radiosensitizing effects of h-R3 with different time series of administration on human lung cancer cells expressing various EGFR levels［J］. Chinese Journal of Clinical Oncology, 2011,(17):1019-1023. (in Chinese)
袁智勇, 刘晓斌, 庄洪卿, 等. 尼妥珠单抗不同给药时序对人肺癌细胞系的放射增敏作用［J］. 中国肿瘤临床, 2011,(17):1019-1023.
［9］Zhang XY, Wu H, Lin X, et al. Scavenging effect of dihydromyricetin on the free radicals by ESR ［J］. Modern Food Science and Technology, 2010,26(10):1040-1042,1070.
张晓元, 吴晖, 林霞, 等. 用ESR检测二氢杨梅素对自由基的清除作用的研究［J］. 现代食品科技, 2010,26(10):1040-1042,1070.
［10］Gu XZh, Yin WB, Yu ZH, et al. Radiation Oncology［M］. 4th ed. Peking: China Union Medical College Press, 2007: 227-228. (in Chinese)
谷铣之, 殷蔚伯, 余子豪, 等. 肿瘤放射治疗学［M］. 第4版, 北京: 中国协和医科大学出版社, 2007: 227-228.
［11］Song MJ, Liu H, Shen YL. The research progress of flavonoids radiosensitization agents［J］. Chinese Journal of Hospital Pharmacy, 2013, (12): 997-999, 1002. (in Chinese)宋梦姣, 刘宏, 申玉莉. 黄酮类放射增敏剂的研究进展［J］. 中国医院药学杂志, 2013, (12): 997-999, 1002.
［12］Hermann RM, Fest J, Christiansen H, et al. Radiosensitization dependent on p53 function in bronchial carcinoma cells by the isoflavone genistein and estradiol in vitro［J］. Strahlenther Onkol, 2007,183(4):195-202.