摘要
目的:观察藤梨根对人结肠癌RKO细胞悬浮生长及失巢凋亡作用。方法:200～800 mg.L-1藤梨根作用于RKO细胞,CCK-8(CCK-8)法检测藤梨根对RKO细胞悬浮生长作用,双嵌入剂乙锭均二聚物(EthD-1)染色检测失巢凋亡,比色法检测半胱天冬酶-3(caspase-3)活性,2-7-二氯氢化荧光素乙二脂染色法结合荧光酶标仪检测细胞内活性氧的产生。结果:200～800 mg.L-1藤梨根可显著抑制RKO悬浮生长,呈剂量依赖性(P<0.01)。200～800 mg.L-1藤梨根作用后悬浮生长RKO细胞吸收EthD-1散发红色荧光,部分细胞可见核碎裂,呈现凋亡形态改变;藤梨根组EthD-1荧光吸光度与对照组比较差异显著(P<0.01)。终质量浓度200～800 mg.L-1藤梨根可显著增强RKO细胞caspase-3活性以及细胞内活性氧的产生(P<0.01)。结论:藤梨根可抑制RKO细胞悬浮生长,促使RKO细胞发生失巢凋亡,可能与活化caspase-3以及升高细胞内活性氧水平相关。
Objective： To observe the effects of Tengligen （root of Actinidia chinensis Planch） on cell growth in suspension and anoikis in human colon carcinoma RKO cells. Method： RKO cells were treated with Tengligen （200-800 mg ·L^-1 ） or same volume of RPMI1640. Cell growth in suspension was detected with CCK-8 assay. Anoikis was visualized by EthD-1 staining. Caspase-3 activities were detected by eolorimetric assay. Intracellular reactive oxygen species （ROS） production was observed by 2＇, 7＇-dichlorofluorescin diacetate staining. Result： 200-800 mg ·L^-1 of Tengligen, significantly inhibited RKO cell growth in suspension in a dose dependent manner （P 〈0.01 ）. After 200-800 mg ·L^-1 of treatment, EthD-1 was absorbed by suspension-cultured RKO ceils to yield a red-fluorescent nuclear staining, and nuclear fragmentation was observed in partial ceils. TheEthD-1 fluorescence in 200-800 mg ·L^-1 of Tengligen treated RKO cells 200-800 mg ·L^-1 of Tengligen also activated caspase-3 and upregulated Conclusion： Tengligen could inhibit RKO cell growth in suspension, were compared with controls （P 〈 0.01 ） intracellular ROS generation （P 〈 0. 01 ） induce anoikis in RKO cells, and mayassociate with caspase-3 activation and ROS production.
出处
《中国实验方剂学杂志》
CAS
北大核心
2013年第16期242-245,共4页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81273726)
上海市西部开发科技合作项目(11495801300)
上海市中医药事业三年行动计划首批重大研究项目(ZYSNXD-CC-ZDYJ017)
国家中医管理局中医肿瘤重点学科合作项目(LHZLK-1107)
上海中医药大学杏林学者项目
龙华医院国家中医临床研究基地“龙医团队、龙医学者”项目(LYTD-04)
关键词
结肠癌
藤梨根
失巢凋亡
半胱天冬酶-3
活性氧
colon carcinoma cell
root of Actinidia chinensis
anoikis
caspases-3
reactive oxygenspecies
