摘要
目的研究当归挥发油(VOAS)对人结肠癌SW480细胞凋亡和自噬的影响及其分子机制。方法将SW480细胞分为6组:空白对照组(0μg·mL^(-1))和5个浓度(VOAS:6.25,12.5,25.0,50.0和100.0μg·mL^(-1))实验组(命名为实验-1、-2,-3,-4和-5组),培养48 h。用噻唑蓝比色法检测VOAS抑制SW480细胞增殖的作用,用流式细胞术和吖啶橙(AO)荧光染色分别观察25, 50μg·mL^(-1)的VOAS作用于SW480细胞后细胞的凋亡和自噬情况,蛋白质印迹法测定细胞凋亡和自噬相关蛋白及磷脂酰肌醇3-激酶/蛋白激酶B/西罗莫司靶蛋白(PI3K/Akt/mTOR)信号转导通路关键蛋白表达水平。结果空白对照组和实验-1、-2,-3,-4和-5组的细胞抑制率分别为0,(14.07±1.96)%,(19.87±3.42)%,(37.09±2.06)%,(49.15±2.83)%和(77.82±2.48)%,IC;为(49.26±2.33)μg·mL^(-1),表明VOAS能够明显抑制SW480细胞的增殖。空白对照组、实验-3组和实验-4组的细胞凋亡率分别为(6.22±1.28)%,(15.32±3.11)%和(35.68±3.89)%;这3组的凋亡相关蛋白Cleaved-Caspase-3的相对表达量分别为(50.05±2.78)%,(74.10±5.80)%和(114.55±5.26)%;这3组的自噬相关蛋白LC3B-Ⅱ的相对表达量分别为(10.56±2.33)%,(51.11±2.82)%和(83.93±10.15)%;这3组的p-mTOR的相对表达量分别为(111.40±3.97)%,(76.37±6.52)%和(59.52±1.99)%。实验-3组和实验-4组的上述指标与空白对照组比较,差异均有统计学意义(均P<0.01);同时,实验-3组和实验-4组与空白对照组比较,细胞内红色荧光强度明显增强。结论 VOAS能够抑制人结肠癌SW480细胞增殖,诱导细胞凋亡及自噬,其机制可能与抑制PI3K/Akt/mTOR信号转导通路有关。
Objective To study the effect and mechanism of volatile oil of angelica (VOAS) on apoptosis and autophagy in human colorectal cancer SW480 cells.Methods Human colorectal cancer SW480 cells were divided into six groups:Blank control group (0μg·m L^(-1)) and experimental groups treated with various concentrations (6.25,12.5,25,50 and 100μg·m L^(-1);named experimental-1,-2,-3,-4,-5groups) of VOAS for 48 h.The inhibitory rate of cell proliferation was tested by MTT assay.The effect of VOAS (25,50μg·m L^(-1)) on apoptosis and autophagy were observed respectively by flow cytometry and acridine orange (AO) staining.The expression of apoptosis-related protein,autophagy hallmark protein and the phosphatidylinositol 3 kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/m TOR) signal pathway were detected by Western blotting.Results The inhibition rates of SW480 cells in the blank control group and experimental-1,-2,-3,-4,-5 groups were 0,(14.07±1.96)%,(19.87±3.42)%,(37.09±2.06)%,(49.15±2.83)%and (77.82±2.48)%,the value of IC;was (49.26±2.33)μg·m L^(-1),6.25-100.00μg·m L^(-1)VOAS could significantly inhibit the proliferation of SW480 cells.The apoptosis rate in the blank control group and experimental-3,-4 groups were (6.22±1.28)%,(15.32±3.11)%,(35.68±3.89)%,respectively;Cleaved-caspase-3 protein levels in the three groups were (50.05±2.78)%,(74.10±5.80)%,(114.55±5.26)%,respectively;LC3B-Ⅱprotein levels in the three groups were (10.56±2.33)%,(51.11±2.82)%,(83.93±10.15)%,respectively;p-m TOR protein levels in the three groups were (111.40±3.97)%,(76.37±6.52)%,(59.52±1.99)%.Compared between experimental-3,-4 groups and blank control group,the differences of the factors were significant(all P<0.01);while the red fluorescence intensity was significantly increased in the cells of treated with VOAS.Conclusion VOAS may induce apoptosis and autophagy in human colorectal cancer sw480 cells through inhibiting PI3K/Akt/m TOR signal transduction pathway.
作者
朱丽娟
罗建云
宋润泽
王丽娟
张安平
臧凯宏
ZHU Li-juan;LUO Jian-yun;SONG Run-ze;WANG Li-juan;ZHANG An-ping;ZANG Kai-hong(College of Pharmacy,Gansu University of Traditional Chinese Medcine(TCM),Lanzhou 730000,Gansu Province,China;Gansu Province Key Laboratory of TCM Pharmacology and Toxicology,Lanzhou 730000,Gansu Province,China;Xi'an Healthcare Security Administration,Xi'an 710021,Shaanxi Province,China;Department of Vascular Surgery,Lanzhou University Second Hospital,Lanzhou 730000,Gansu Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2021年第23期3253-3256,共4页
The Chinese Journal of Clinical Pharmacology
基金
甘肃省高等学校科研基金资助项目(2018-A050)
甘肃省中药药理与毒理学重点实验室开放基金资助项目(ZDSYS-KJ-2018-007)。
