摘要
目的:探讨敲低BCLAF1对肝癌SMMC-7221细胞增殖、侵袭及耐药性的影响。方法:构建shRNA-BCLAF1载体,按照空白对照组、shRNA-NC(阴性对照)组、shRNA-BCLAF1组转染SMMC-7221细胞;分别采用qPCR及Western blot鉴定干扰效果;MTT法检测各组细胞增殖活性;Transwell检测各组细胞侵袭能力;划痕实验检测各组细胞迁移能力;使用0、4、8、12、16μmol/L的索拉非尼对各组细胞进行处理,比较细胞活性;Western blot法检测各组16μmol/L索拉非尼处理后PI3K、p-PI3K、Akt、p-Akt蛋白表达水平。结果:shRNA-BCLAF1组BCLAF1基因mRNA及蛋白相对表达量均低于空白对照组、shRNA-NC组(P<0.05);常规培养48 h后,shRNA-BCLAF1组SMMC-7221细胞增殖率低于空白对照组及shRNA-NC组(P<0.05);shRNA-BCLAF1组SMMC-7221细胞侵袭率、迁移率低于空白对照组及shRNA-NC组(P<0.05);shRNA-BCLAF1组各浓度索拉非尼处理条件下SMMC-7221细胞活性低于空白对照组及shRNA-NC组(P<0.05);16μmol/L索拉非尼处理后,shRNA-BCLAF1组p-PI3K、p-Akt蛋白相对表达量低于空白对照组及shRNA-NC组(P<0.05)。结论:敲低BCLAF1能抑制肝癌SMMC-7221细胞增殖、侵袭,降低对索拉非尼的耐药性,机制可能与抑制PI3K/Akt信号通路有关。
Objective:To explore the effects of knocking down BCLAF1 on the proliferation,invasion and drug resistance of liver cancer SMMC-7221 cells.Methods:The shRNA-BCLAF1 vector was constructed.The SMMC-7221 cells in blank control group,shRNA-NC(negative control)group and shRNA-BCLAF1 group were transfected.The interference effectswere identified by qPCR and Western blot.The proliferation activity of cells in each group was detected by MTT.The invasion ability of cells in each group was detected by Transwell.The migration ability of cells in each group was detected by scratch assay.Cells in each group were treated with sorafenib(0,4,8,12,16μmol/L).The cells viability was compared.The expression levels of PI3K,p-PI3K,Akt and p-Akt proteins after treatment with 16μmol/L sorafenib in each group were detected by Western blot.Results:The relative expression levels of BCLAF1 gene and proteins in shRNA-BCLAF1 group were significantly lower than those in blank control group and shRNA-NC group(P<0.05).After 48 h of routine culture,proliferation rate of SMMC-7221 cells in shRNA-BCLAF1 group was significantly lower than that in blank control group and shRNA-NC group(P<0.05).The invasion rate and migration rate of SMMC-7221 cells in shRNA-BCLAF1 group were significantly lower than those in blank control group and shRNA-NC group(P<0.05).The viability of SMMC-7221 cells treated with different concentrations of sorafenib in shRNA-BCLAF1 group was significantly lower than that in blank control group and shRNA-NC group(P<0.05).After treatment with 16μmol/L sorafenib,relative expression levels of p-PI3K and p-Akt proteins in shRNA-BCLAF1 group were significantly lower than those in blank control group and shRNA-NC group(P<0.05).Conclusion:Knocking downBCLAF1 can inhibit the proliferation and invasion of liver cancer SMMC-7221 cells,and reduce their resistance to sorafenib.The mechanism may be related to inhibiting PI3K/Akt signaling pathway.
作者
朱朝玉
范长儒
张玉良
ZHU Chao-yu;FAN Chang-ru;ZHANG Yu-liang(Department 2 of General Surgery,Linyi Cancer Hospital,Linyi 276034,Shandong,China)
出处
《川北医学院学报》
CAS
2022年第2期147-152,共6页
Journal of North Sichuan Medical College
关键词
BCLAF1
肝癌
增殖
侵袭
耐药性
BCLAF1
Liver cancer
Proliferation
Invasion
Drug resistance
