白花蛇舌草提取物对肝癌细胞的放疗增敏作用观察  

Radiosensitizing effect of extract of Hedyotis diffusa Willd on liver cancer cell lines

作　　者：郭驹 金凯 李美乐 唐婷 谢裕安 GUO Ju;JIN Kai;LI Meile;TANG Ting;XIE Yu'an(Graduate School,Guangxi University of Chinese Medicine,Nanning 530200,China;不详)

机构地区：[1]广西中医药大学研究生院,南宁530200 [2]广西生殖健康与出生缺陷防治重点实验室 [3]广西医科大学研究生院 [4]广西医科大学附属肿瘤医院实验研究部 [5]广西壮族自治区妇幼保健院

出　　处：《山东医药》2022年第33期1-4,共4页Shandong Medical Journal

基　　金：国家重点研发计划项目(NO2019YFC0121901);广西重点实验室运行补助项目(21-220-22)。

摘　　要：目的观察白花蛇舌草提取物(EHDW)对人肝癌细胞系Huh-7、SMMC-7721的放疗增敏作用。方法将Huh-7、SMMC-7721细胞分别分成IR组和IR+EHDW组,IR+EHDW组加入40μg/mL的EHDW培养液,IR组加入同等体积的细胞培养液,继续培养12 h后,两组分别给予0、2、4、6、8 Gy照射剂量的X射线照射5 min,采用平板克隆实验观察EHDW对Huh-7、SMMC-7721细胞增殖能力的影响,以细胞存活分数(SF)表示;采用GraphPad Prism 9.0软件利用单击多靶模型和L-Q模型拟合生存曲线,并使用Sigma Plot V14.5软件计算出各组细胞的放射生物学参数,包括外推数(N)、平均致死剂量(D_(0))、准阈剂量(Dq)、2 Gy剂量照射时的细胞存活分数(SF_(2))、放疗增敏比(SER)。取Huh-7、SMMC-7721细胞分成对照组、IR组、EHDW组、IR+EHDW组,EHDW组和IR+EHDW组分别加入40μg/mL的EHDW培养液,对照组和IR组加入同等体积的细胞培养液,各组细胞继续培养12 h后,IR组和IR+EHDW组给予8 Gy照射剂量的X射线照射5 min,辐照结束后继续培养24 h后更换培养基,48 h后开始收样,采用流式细胞术测算Huh-7、SMMC-7721细胞凋亡率。结果不同照射剂量下,IR+EHDW组Huh-7、SMMC-7721细胞存活分数均低于IR组(P均<0.05)。IR+EHDW组Huh-7、SMMC-7721细胞的SER分别为1.324、1.281。对照组、IR组、EHDW组、IR+EHDW组Huh-7细胞凋亡率分别为20.75%±1.48%、54.27%±1.58%、63.52%±1.76%、75.97%±2.02%,组间相比,P均<0.05。对照组、IR组、EHDW组、IR+EHDW组SMMC-7721细胞凋亡率分别为24.69%±2.18%、35.95%±1.19%、52.66%±1.04%、77.77%±0.89%,组间相比,P均<0.05。结论EHDW可抑制Huh-7、SMMC-7721细胞增殖,增强其放疗敏感性,并诱导细胞凋亡,EHDW对人肝癌细胞系Huh-7、SMMC-7721具有放疗增敏作用。Objective To observe the radiosensitizing effect of extract of hedyotis diffusa willd(EHDW)on human liver cancer cell lines Huh-7 and SMMC-7721.Methods Huh-7 and SMMC-7721 cells were divided into the IR and IR+EHDW groups,respectively.EHDW culture medium(40μg/mL)was added to the IR+EHDW group and the same volume of cell culture medium was added to the IR group,and the cells were continued to be cultured for 12 h.After the two groups were given X-ray irradiation at 0,2,4,6 and 8 Gy irradiation doses for 5 min,and the plate cloning assay was used to observe the effect of EHDW on the proliferation ability of Huh-7 and SMMC-7721 cells which was expressed as cell survival fraction(SF).GraphPad Prism 9.0 software was used to fit the survival curves using one-click multi-target model and L-Q model,and the radiobiological parameters of each group were calculated using Sigma Plot V14.5 software,including extrapolation number(N),mean lethal dose(D_(0)),quasi-threshold dose(Dq),cell survival fraction at 2 Gy dose(SF2)irradiation,and radiotherapy sensitization enhancement ratio(SER).Huh-7 and SMMC-7721 cells were divided into the control,IR,EHDW and IR+EHDW groups.EHDW culture medium(40μg/mL)was added to the EHDW and IR+EHDW groups,and the same volume of cell culture medium was added to the control and IR groups,and the cells in each group were continued to be cultured for 12 h.After the cells in the IR and IR+EHDW groups were given 8 Gy irradiation,we changed the culture medium after 24 h.After 48 h,the cells were collected,and the apoptosis rates of Huh-7 and SMMC-7721 cells were calculated by flow cytometry.Results The survival fraction of Huh-7 and SMMC-7721 cells in the IR+EHDW group was lower than that in the IR group at different irradiation doses(P<0.05),and the SERs of Huh-7 and SMMC-7721 cells in the IR+EHDW group were 1.324 and 1.281,respectively.The apoptosis rates were 20.75%±1.48%,54.27%±1.58%,63.52%±1.76%,and 75.97%±2.02%in the control,IR,EHDW,and IR+EHDW groups,respectively,with statistically significan

关 键 词：放疗增敏剂 白花蛇舌草 白花蛇舌草提取物 肝癌 HUH-7细胞 SMMC-7721细胞 放射疗法 

分 类 号：R735.7[医药卫生—肿瘤]