长链非编码RNA AURKAPS1通过上调RAC1活化细胞外信号调节激酶信号通路促进肝癌进展的机制研究  

Mechanism of long non-coding RNA AURKAPS1 promoting the progression of hepatocellular carcinoma by up regulating RAC1 activated extracellular signal-regulated kinase signaling pathway

作　　者：赵静[1] 樊洁净 冯娟娟[1] 郑守华[2] 李建华[2] Zhao Jing;Fan Jiejing;Feng Juanjuan;Zheng Shouhua;Li Jianghua(Department of Surgery,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Department of General Surgery,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院外科医学部,450052 [2]郑州大学第一附属医院普通外科,450052

出　　处：《中华实验外科杂志》2022年第10期1864-1868,共5页Chinese Journal of Experimental Surgery

基　　金：河南省自然科学基金面上项目(202300410451)。

摘　　要：目的探讨AURKAPS1在肝癌细胞中的表达、功能和分子机制,为肝癌的防治提供新思路。方法采用实时荧光定量聚合酶链反应(PCR)技术对肝癌细胞系和正常肝脏上皮细胞进行检测;将肝癌细胞系随机分为空白组,AURKAPS1组、RAC1组和AURKAPS1+RAC1组,其中空白组表示肝癌细胞不进行任何干预,AURKAPS1组中的肝癌细胞仅接受AURKAPS1干预,RAC1组为转染RAC1的肝癌细胞,AURKAPS1+RAC1组表示同时转染AURKAPS1和RAC1,采用实时荧光定量PCR技术检测4组细胞中RAC1的表达;采用细胞计数试剂盒(CCK-8)法、流式细胞术和Transwell转移法检测肿瘤细胞的增殖、凋亡和迁移能力;蛋白质免疫印迹法测定各组细胞中细胞外信号调节激酶(ERK)和RAC1蛋白表达;间接免疫荧光检测细胞中ERK的表达。组间独立样本采用t检验,多组间进行F检验、LSD检验。结果肝癌组织中RAC1 mRNA的表达量(0.98±0.08)显著高于正常人肝上皮细胞组织(0.72±0.03),差异有统计学意义(t=30.431,P<0.05);AURKAPS1组(1.03±0.10)、RAC1组(1.05±0.09)及AURKAPS1+RAC1组(1.16±0.07)的RAC1 mRNA的表达量均高于空白组(1.16±0.07),AURKAPS1+RAC1组的RAC1 mRNA表达量也显著高于分别转染AURKAPS1、RAC1的肝癌细胞组,差异有统计学意义(F=5.991,P<0.05);CCK-8法检测肝癌细胞的增殖,随着时间的增加,除空白组,其余3组的吸光度(A)_(450)值均增加,96 h后AURKAPS1+RAC1组的A_(450)值显著高于RAC1组及AURKAPS1组(F_(0 h)=2.941,F_(24 h)=26.454,F_(48 h)=104.60,F_(72 h)=319.555,F_(96 h)=564.65)P<0.05;AURKAPS1组[(5.89±3.82)%]、RAC1组[(5.81±3.85)%]及AURKAPS1+RAC1组[(4.23±4.15)%]的总凋亡率均低于空白组[(6.75±3.36)%],且AURKAPS1+RAC1组的总凋亡率显著低于RAC1、AURKAPS1组,F=77.369,P<0.05;AURKAPS1组(131.24±9.25)、RAC1组(129.87±9.41)及AURKAPS1+RAC1组(153.62±7.48)的穿膜细胞数均高于空白组(117.64±10.53),且AURKAPS1+RAC1组的穿膜细胞数显著高于RAC1组及AURKAPS1组(F=91.118,P<0.05);Western bloObjective To explore the expression,function and molecular mechanism of AURKAPS1 in hepatocellular carcinoma cells,and provide a new idea for the prevention and treatment of hepatocellular carcinoma.Methods Real-time fluorescent quantitative polymerase chain reaction(PCR)was used to detect hepatoma cell lines and normal liver epithelial cells;The liver cancer cell line was randomly divided into blank group,AURKAPS1 group,RAC1 group and AURKAPS1+RAC1 group.The blank group indicated that the liver cancer cells did not undergo any intervention.The liver cancer cells in AURKAPS1 group only received AURKAPS1 intervention.RAC1 group was the liver cancer cells transfected with RAC1.AURKAPS1+RAC1 group showed that efw was transfected with AURKAPS1 and RAC1 at the same time.The expression of RAC1 in the four groups of cells was detected by real-time fluorescent quantitative PCR technology;cell counting kit-8(CCK-8),flow cytometry and Transwell metastasis were used to detect the proliferation,apoptosis and migration of tumor cells;The expression of extracellular signal-regulated kinase(ERK)and RAC1 protein was detected by Western blotting;The expression of ERK was detected by indirect immunofluorescence.SPSS 26.0 analyzed the data,RAC1,ERK expression,±s means independent sample t test between groups,F test and LSD test between groups,P<0.05,which indicates that the comparison between groups is significant.Results The expression of RAC1 mRNA in hepatocellular carcinoma(0.98±0.08)was significantly higher than that in normal human liver epithelial cells(0.72±0.03),and the difference was statistically significant(t=30.431 P<0.05);AURKAPS1 group(1.03±0.10),RAC1 group(1.05±0.09)The expression of RAC1 mRNA in the AURKAPS1+RAC1 group was higher than that in the blank group(1.16±0.07);CK-8 method was used to detect the proliferation of liver cancer cells.With the increase of time,the A_(450) value of the other three groups increased,except the blank group.After 96 h,the A_(450) value of AURKAPS1+RAC1 group was significantly h

关 键 词：长链非编码RNA AURKAPS1 Ras相关的C3肉毒杆菌毒素底物1 细胞外信号调节激酶信号通路 肝癌细胞 实时荧光定量聚合酶链反应 

分 类 号：R735.7[医药卫生—肿瘤]