摘要
目的:观察组蛋白去乙酰化酶抑制剂(HDACi)丙戊酸(VPA)单独或联合X射线照射对人三阴性乳腺癌(TNBC)MDA-MB-231细胞增殖的抑制作用,并分析联合照射对MDA-MB-231细胞凋亡和自噬的影响。方法:将处于对数生长期的MDA-MB-231细胞分为对照组、不同浓度(2.5、5.0、10.0、20.0和40.0 mmol·L^(-1))VPA组、4 Gy X射线照射组和不同浓度(2.5、5.0、10.0、20.0和40.0 mmol·L^(-1))VPA联合4 Gy X射线照射组。于作用24、48和72 h后,采用CCK-8法检测各组MDA-MB-231细胞增殖率。MDA-MB-231细胞分为对照组、4 Gy X射线照射组、5 mmol·L^(-1) VPA组、5 mmol·L^(-1) VPA联合4 Gy X射线照射组。于作用24和48 h后,流式细胞术检测各组MDA-MB-231细胞凋亡率和自噬率。结果:与对照组比较,VPA组和VPA联合4 Gy X射线照射组MDA-MB-231细胞增殖率均明显降低(P<0.01),且呈浓度和时间依赖性;与4 Gy X射线照射组比较,VPA联合4 Gy X射线照射组MDA-MB-231细胞增殖率均明显降低(P<0.01)。作用24 h后,2.5 mmol·L^(-1) VPA联合4 Gy X射线照射组MDA-MB-231细胞增殖率低于2.5 mmol·L^(-1) VPA组(P<0.05);作用72 h后,2.5和10.0 mmol·L^(-1) VPA联合4 Gy X射线照射组MDA-MB-231细胞增殖率低于2.5和10.0 mmol·L^(-1)VPA组(P<0.05)。5.0 mmol·L^(-1)VPA单独或联合4 Gy X射线作用MDA-MB-231细胞24和48 h后,与对照组比较,4 Gy X射线照射组、5.0 mmol·L^(-1)VPA组和5.0 mmol·L^(-1)VPA联合4 Gy X射线照射组MDA-MB-231细胞凋亡率和自噬率均明显升高(P<0.05或P<0.01)。与4 Gy X射线照射组比较,5.0 mmol·L^(-1)VPA联合4 Gy X射线照射组MDA-MB-231细胞凋亡率和自噬率均明显升高(P<0.01)。作用48 h后,与5.0 mmol·L^(-1)VPA组比较,5.0 mmol·L^(-1) VPA联合4 Gy X射线照射组MDA-MB-231细胞自噬率升高(P<0.05)。结论:VPA对MDA-MB-231细胞具有增殖抑制作用,并呈现浓度和时间依赖性,且VPA和X射线的联合抑制效果更优,其机制可能是二者联合引起细胞凋亡和自噬增加。
Objective:To observe the inhibitory effect of valproic acid(VPA),a histone deacetylase inhibitor(HDACi),on the proliferation of the human triple negative breast cancer(TNBC)MDA-MB-231 cells alone or combined with X-ray irradiation,and to analyze the effect of combined irradiation on the apoptosis and autophage of the MDA-MB-231 cells.Methods:The MDA-MB-231 cells at logarithmic growth phase were divided into control group,different concentrations(2.5,5.0,10.0,20.0,and 40.0 mmol·L^(-1))of VPA groups 4 Gy,X-ray irradiation groups,and different concentrations(2.5,5.0,10.0,20.0,and 40.0 mmol·L^(-1))of VPA combined with 4 Gy X-ray irradiation groups.The proliferation rates of the cells in various groups were detected by CCK-8 method after treated for 24,48,and 72 h.The cells were divided into control group,4 Gy X-ray irradiation group,5 mmol·L^(-1) VPA group,5 mmol·L^(-1) VPA combined with 4 Gy X-ray irradiation group,the apoptotic rates and autophage rates of the cells in various groups were analyzed by fiow cytometry after treated for 24 and 48 h.Results:Compared with control group,the proliferation rates of the MDA-MB-231 cells in VPA group and VPA combined with 4 Gy X-ray irradiation groups were decreased significantly in a concentration and time-dependent manner(P<0.01).Compared with 4 Gy X-ray irradiation group,the proliferation rates of the MDA-MB-231cells in VPA combined with 4 Gy X-ray irradiation groups were significantly decreased(P<0.01).Compared with 2.5 mmol·L^(-1) VPA group,the proliferation rate of the MDA-MB-231 cells in 2.5 mmol·L^(-1) VPA combined with 4 Gy X-ray irradiation group after treated for 24 h was decreased(P<0.05).After treated for 72 h,the proliferation rates of the MDA-MB-231 cells in 2.5 and 10.0 mmol·L^(-1) VPA combined with 4 Gy X-ray irradiation groups were lower than those in 2.5 and 10.0 mmol·L^(-1) VPA groups(P<0.05),respectively.After treated for 24 and 48 h,the apoptotic rates and autophagy rates of the MDA-MB-231 cells in 4 Gy X-ray group,5 mmol·L^(-1) VPA group and 5 mmol·L^(-1) VPA combined with 4 Gy X-ray group were significantly higher than those in control group(P<0.05 or P<0.01).Compared with 4 Gy X-ray irradiation group,the apoptotic rate and autophagy rate of the MDA-MB-231 cells in 5 mmol·L^(-1) VPA combined with 4 Gy X-ray irradiation group were significantly increased(P<0.01).After treated for 48 h,compared with 5 mmol·L^(-1) VPA group,the autophagy rate of the MDA-MB-231 cells in 5 mmol·L^(-1) VPA combined with 4 Gy X-ray irradiation group was increased(P<0.05).Conclusion:VPA can inhibit the proliferation of MDA-MB-231 cells in a concentration and time-dependent manner,and the combined inhibitory effect of VPA and X-ray is better,and the mechanism may be involved in the increasing of apoptosis and autophage of the MDA-MB-231 cells induced by the combination of VPA and X-ray.
作者
李官虎
郎庆旭
刘纯岩
刘沁
耿梦柔
李晓倩
王珍琦
LI Guanhu;LANG Qingxu;LIU Chunyan;LIU Qin;GENG Mengrou;LI Xiaoqian;WANG Zhenqi(School of Public Health,Jilin University,Key Laboratory of Radiobiology of National Health Commission,Changchun 130021,China;Department of Radiology,Second Hospital,Jilin University,Changchun 130041,China)
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2022年第3期622-629,共8页
Journal of Jilin University：Medicine Edition
基金
吉林省卫生健康科技能力提升项目(2021JC039)
吉林省科技厅科技发展计划项目(20160101227JC)
吉林省教育厅“十三五”科学技术项目(JJKH20170826KJ)
吉林大学“大学生创新创业训练计划”项目(202010183443)。
关键词
组蛋白去乙酰化酶抑制剂
丙戊酸
细胞增殖
细胞凋亡
自噬
Histone deacetylase inhibitor
Valproic acid
Cell proliferation inhibition
cell apoptosis
Autophage
作者简介
李官虎(1991-),男,吉林省延吉市人,在读硕士研究生,主要从事辐射肿瘤学方面的研究;通信作者:王珍琦,副教授,硕士研究生导师(E-mail:wangzq@jlu.edu.cn)。
