摘要
目的考察骆驼刺中有效成分tortoside A(TOA)对人宫颈癌He La细胞增殖和凋亡的影响及可能机制。方法CCK-8法检测不同浓度TOA(3.125、6.25、12.5、25、50、100 mg·L^(-1))在不同处理时间(24,48 h)对He La细胞增殖的抑制作用,并根据细胞存活率计算IC_(50)值。将TOA干预He La细胞48 h的1/2 IC_(50)和1/4 IC_(50)值设为高、低剂量,顺铂对He La细胞的1/2 IC_(50)值设为阳性对照,另设空白对照,以Hoechst染色法、流式细胞术检测TOA对He La细胞凋亡的影响,流式细胞术检测细胞周期及线粒体膜电位变化,RT-PCR和Western blot法检测TOA对He La细胞凋亡相关蛋白和基因表达的影响。结果TOA对He La细胞增殖的抑制率有浓度和时间依赖性趋势,干预48 h的IC_(50)值为21.3μg·m L^(-1)。与空白对照组比较,顺铂1.15μg·m L^(-1)组与TOA 10.65μg·m L^(-1)、5.33μg·m L^(-1)组He La细胞G;/G;期百分比增加、克隆形成数减少、细胞凋亡率增加、线粒体膜电位下降,caspase-3、caspase-7和PARP蛋白和基因表达显著增加,而Bcl-2的表达显著降低(P<0.05,P<0.01)。结论TOA对He La细胞的增殖具有抑制作用,其抑制作用可能与促进细胞凋亡有关。
AIM To study the effects and possible mechanism of tortoside A(TOA),an effective component of Alhagi sparsifolia Shap.,on proliferation and apoptosis of human cervical cancer He La cells.METHODS The CCK-8method was used to detect the effect of different concentrations of TOA(3.125,6.25,12.5,25,50,100 mg·L^(-1))on He La cells proliferation compared with blank control after 24 h or 48 h of culture,and IC_(50) was calculated based on cell survival.The 1/2 IC_(50) and 1/4 IC_(50) of 48 h treated He La cells with TOA were used as the high and low dose of TOA groups,the positive control group was 1/2 IC_(50) of cisplatin on He La cells,and the blank control group was set up.Then,Hoechst staining and flow cytometry were used to detect the effect of TOA on He La cell apoptosis.Flow cytometry was used to detect the changes of cell cycle and mitochondrial membrane potential.RT-PCR and Western blot were used to detect the effect of TOA on the expression of apoptosis related proteins and genes in He La cells.RESULTS The inhibition rate of TOA on He La cell proliferation showed a concentration and time dependent trend,and the IC_(50) value after 48 h of intervention was 21.3μg·m L^(-1).Compared with the blank control group,cisplatin 1.15μg·m L^(-1)and TOA 10.65μg·m L^(-1)and 5.33μg·m L^(-1)increased the percentage of G;/G;phase,decreased the number of clone formation,increased the apoptosis rate,and decreased the mitochondrial membrane potential,meanwhile they increased the protein and gene expression of caspase-3,caspase-7 and PARP and decreased the expression of Bcl-2(P<0.05,P<0.01).CONCLUSION TOA has an inhibitory effect on the proliferation of He La cells,which may be related to the promotion of cell apoptosis.
作者
马晓玲
刘雪松
柴丽华
陈刚
石磊岭
魏鸿雁
MA Xiao-ling;LIU Xue-song;CHAI Li-hua;CHEN Gang;SHI Lei-ling;WEI Hong-yan(Xinjiang Institute of Chinese Medicine and Medicine of National,Urumqi XINJIANG 830002,China;College of Life Sciences and Technology,Xinjiang University,Urumqi XINJIANG 830046,China;Youcare Pharmaceutical Group Co.,Ltd.,BEIJING 100176,China)
出处
《中国新药与临床杂志》
CAS
CSCD
北大核心
2022年第2期110-116,共7页
Chinese Journal of New Drugs and Clinical Remedies
基金
新疆自治区自然科学基金项目(2018D01B12)。
