丁酸钠对大肠癌HCT-116细胞增殖及自噬影响研究  

Role of autophagy in sodium butyrate-induced death of human colorectal cancer HCT-116 cells

作　　者：谢文权[1] 陈嘉定 李程[1] 陈晓洋[1] 张锦涛[1] 伊曼[1] 孙素霞[1] 

机构地区：[1]南方医科大学公共卫生与热带医学学院营养与食品卫生学系,广东广州510515

出　　处：《中华肿瘤防治杂志》2015年第5期340-343,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(81202204);广东省自然科学基金(S2012010009467);广东省大学生创新训练项目(201412121107);南方医科大学优秀青年教师项目

摘　　要：目的研究丁酸钠(sodium butyrate,NaB)对大肠癌HCT-16细胞增殖和自噬的影响。方法不同浓度(0.1、0.5、1、2和5mmol/L)NaB处理大肠癌HCT-116细胞,MTT法检测细胞的存活率;蛋白质印迹法检测自噬特异性蛋白LC3B表达量的变化;单丹磺酰尸胺(MDC)和吖啶橙(AO)染色检测酸性自噬泡的形成。结果单因素方差分析结果显示,不同浓度(0.1、0.5、1、2和5mmol/L)NaB处理组与对照组细胞的存活率相比,差异有统计学意义,F=24.076,P<0.001;0.5mmol/L浓度的NaB作用24h即能显著抑制大肠癌HCT-116细胞的增殖,细胞存活率为(94.00±4.09)%,P<0.05;1、2、5mmol/L浓度的NaB作用大肠癌HCT-116细胞24h,细胞存活率分别为(89.43±1.83)%、(84.56±4.77)%和(81.00±7.38)%,与对照组比较,P值均<0.01。2mmol/L NaB处理24后,HCT-116细胞内酸性囊泡细胞器明显蓄积;不同浓度NaB处理HCT-116细胞24h后,LC3B-Ⅱ蛋白的表达量随着NaB浓度增加而增加;2 mmol/L NaB处理HCT-116不同时间(10min、30min、1h、2h、4h、6h、12h和24h)后,LC3B-Ⅱ蛋白的表达量随着NaB作用时间的延长而增加。结论 NaB可抑制大肠癌HCT-116细胞增殖并诱导其发生自噬。OBJECTIVE To investigate the effects of sodium butyrate （NAB） on human colorectal cancer HCT-116 cell proliferation and autophagy. METHODS HCT-116 cells were treated with different doses（0.1.0.5.1.2.5 mmol/L） of NaB for 24 h,cell viability was assessed by methyl thiazolyl tetrazolium（MTT） assay. Autophagosomes was observed using fluorescent microscope by acridine orange（AO） and monodansylcadaverine（MDC） staining. The autophagy-specific protein LC3B （microtubule- associated protein 1 light chain 3） was detected by Western Blot. RESULTS The result of one-way analysis of variance showed that the proliferation of the dose groups had statistically significant difference com- pared with the control group（F=24. 076 ,P〈0. 001）. 0.5 mmol/L NaB resulted in suppression of cell growth and cell vi- ability was （94.00±4.09）% ,P〈0.05; 1.2.5 mmol/L NaB resulted in suppression of cell growth,cell viabilities were （89.43±1.83）%, （84.56±4.77）% and （81.00±7.38）%（P〈0.01）. 2 mmol/L NaB resulted in accumulation of acidic vacuoles in HCT-116 cells. NaB increased the transition of LC3B- I to LC3B- II in HCT-116 cells in a time- and dose-de- pendent manner. CONCLUSION NaB inhibits proliferation and induces autophagy in colorectal cancer cells.

关 键 词：肠肿瘤 丁酸钠 自噬 细胞增殖 印迹法 蛋白质 

分 类 号：R735.34[医药卫生—肿瘤]