摘要
目的探究放线菌素D(Act-D)与沉默外被体包被蛋白β2亚基(COPB2)对肝癌细胞的影响及其作用机制。方法将人肝癌HepG2细胞分为对照组、实验组、si-NC组、si-COPB2组、si-COPB2+NaCl组和si-COPB2+Act-D组。实验组加入200 ng·mL^(-1)Act-D;对照组加入等体积生理盐水;si-NC组和si-COPB2组分别转染阴性对照寡核苷酸和COPB2小干扰RNA,转染成功后,si-COPB2+Act-D组加入200 ng·mL^(-1)Act-D,si-COPB2+NaCl组加入等体积生理盐水。用细胞计数试剂盒-8(CCK-8)检测细胞增殖;用原位末端标记法(TUNEL)检测细胞的凋亡情况;用蛋白质印迹法(Western blot)检测COPB2蛋白的表达。结果对照组、实验组、si-NC组、si-COPB2组、si-COPB2+NaCl组和si-COPB2+Act-D组HepG2细胞72h的增殖抑制率分别为0,(78.48±8.11)%,0,(17.42±1.75)%,(17.61±1.76)%和(79.87±7.99)%;这6组的HepG2细胞凋亡率分别为(7.89±0.96)%,(43.63±4.37)%,(7.91±0.82)%,(11.24±0.12)%,(10.87±1.09)%和(52.34±5.25)%;对照组和实验组COPB2蛋白的表达分别为(1.05±0.11)和(0.24±0.03)。实验组与对照组比较,si-COPB2组与si-NC组比较,si-COPB2+Act-D组与si-COPB2+NaCl组比较,差异均有统计学意义(P<0.01,P<0.001,P<0.05)。结论放线菌素D联合沉默COPB2基因的表达,能够抑制肝癌细胞的增殖,促进细胞的凋亡。
Objective To explore the effects of actinomycin D(Act-D)and silent coatomer protein complex subunit beta 2(COPB2)on liver cancer cells and their mechanism.Methods Human HepG2 cells were divided into control group,experimental group,si-NC group,si-COPB2 group,si-COPB2+NaCl group and si-COPB2+Act-D group.Experiment group were added with 200 ng·mL^(-1)Act-D,the control group was added with the same volume of normal saline as the experiment group;si-NC and the si-COPB2 group were transfected with negative control oligonucleotide and COPB2 small interfering RNA respectively.After the transfection was successful,the si-COPB2+Act-D group was added with 200 ng·mL^(-1)Act-D,si-COPB2+NaCl group Add an equal volume of saline.Cell counting kit-8(CCK-8)was used to detect cell proliferation;in situ end labeling(TUNEL)was used to detect cell apoptosis;Western blot was used to detect the expression of COPB2 protein.Results The cell proliferation rates of the control group,experimental group,si-NC group,si-COPB2 group,si-COPB2+NaCl group and si-COPB2+Act-D group were 0,(78.48±8.11)%,0,(17.42±1.75)%,(17.61±1.76)%and(79.87±7.99)%;HepG2 cell apoptosis rates were(7.89±0.96)%,(43.63±4.37)%,(7.91±0.82)%,(11.24±0.12)%,(10.87±1.09)%and(52.34±5.25)%;the expression of COPB2 protein of control group,experimental group were(1.05±0.11),and(0.24±0.03),respectively.Compared experimental group with control group,si-COPB2 group compared with si-NC group,si-COPB2+Act-D group compared with si-COPB2+NaCl group,the difference were statistically significant(P<0.01;P<0.001;P<0.05).Conclusion Actinomycin D combined with silencing the expression of COPB2 gene can inhibit the proliferation of liver cancer cells and promote cell apoptosis.
作者
汪杨
黄晓梅
肖志华
WANG Yang;HUANG Xiao-mei;XIAO Zhi-hua(Department of Digestive Diseases,The Second Affiliated Hospital of Nanchang University,Jiangxi 330000,Nanchang Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2021年第20期2755-2758,共4页
The Chinese Journal of Clinical Pharmacology
关键词
放线菌素D
肝癌细胞
COPB2基因
凋亡
actinomycin D
hepatocellular carcinoma cells
COPB2 gene
apoptosis
