环状RNA TADA2A在结直肠癌组织的表达及其对结直肠癌细胞增殖、凋亡的影响  

Expression of circular RNA transcriptional adaptor 2A in colorectal cancer tissues and its effect on proliferation and apoptosis of colorectal cancer cells

作　　者：李震[1] 王世豪 孙伟鹏 姚弘虞 谷晓明[1] 段小飞[1] Li Zhen;Wang Shihao;Sun Weipeng;Yao Hongyu;Gu Xiaoming;Duan Xiaofei(Department of Colorectal and Anal 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结直肠肛门外科,450052

出　　处：《中华实验外科杂志》2021年第2期218-221,共4页Chinese Journal of Experimental Surgery

基　　金：2018年度河南省高等学校重点科研项目(18A320058)。

摘　　要：目的观察环状RNA TADA2A在结直肠癌组织中的表达及其对结直肠癌细胞增殖、凋亡的影响。方法实时定量反转录聚合酶链反应(RT-qPCR)法检测58对结直肠癌组织及癌旁组织中circTADA2A的相对表达量,分析其与患者临床病理特征间的关系。构建circTADA2A过表达质粒及对照质粒并分别转染结直肠癌细胞株(人结直肠癌细胞LoVo、HCT116)。采用细胞计数试剂盒(CCK-8)实验及流式细胞术检测过表达circTADA2A对LoVo和HCT116细胞增殖能力、凋亡水平的影响。体内实验:将12只裸鼠采用随机数字表法分为2组,分别皮下注射转染circTADA2A(过表达组)及空载质粒(对照组)的结直肠癌细胞株HCT116,比较两组皮下瘤体积和质量。定性资料采用χ^(2)检验或Fisher’s精确检验。定量资料采用t检验或非参数检验。结果circTADA2A在结直肠癌组织中的相对表达量(1.12±0.45)显著低于癌旁组织(0.48±0.22,t=9.620,P<0.01),且其表达量与与肿瘤T分期(χ^(2)=6.855,P<0.05)、淋巴结转移(χ^(2)=10.952,P<0.05)、TNM分期(χ^(2)=6.023,P<0.05)、肿瘤分化程度(χ^(2)=5.376,P<0.05)显著相关,差异均有统计学意义。CCK-8实验结果表明,过表达组LoVo和HCT116细胞48、72 h时增殖能力均显著低于对照组,48 h过表达组[LoVo为,(0.34±0.02);HCT116为,(0.31±0.03)比对照组LoVo为,(0.64±0.03);HCT116为,(0.53±0.04),tLoVo=11.190,tHCT116=7.510,P值均<0.01];72 h过表达组[LoVo为,(0.71±0.02);HCT116为,(0.60±0.15)比对照组LoVo为,(0.92±0.03);HCT116为,(0.79±0.02),tLoVo=10.060,tHCT116=12.050,P值均<0.01]。流式细胞结果显示,过表达组细胞凋亡率[LoVo为,(23.93±1.51)%;HCT116为,(20.97±2.36)%]均显著高于对照组[LoVo为,(7.60±0.57)%;HCT116为,(6.83±2.36)%,tLoVo=-17.420,tHCT116=-10.210,P值均<0.01],差异均有统计学意义。体内实验结果显示,过表达组皮下肿瘤[体积(500.1±71.0)mm3、质量(488.1±77.6)mg]均显著小于对照组[(940.8±51.6)mm3、(871.0±73.3)mg,t体积=12.280,Objective To investigate the expression of circular RNA transcriptional adaptor 2A(circTADA2A)in colorectal cancer(CRC)tissues and its effect on the proliferation and apoptosis of CRC cells.Methods Real-time quantitative reverse transcriptase-polymerase chain reaction(RT-qPCR)was used to detect the relative expression of circTADA2A in 58 pairs of CRC tissues and adjacent tissues,and the relationship between circTADA2A and the clinicopathological characteristics of patients were analyzed subsequently.The circTADA2A overexpression plasmid and empty vector were constructed and then transfected into CRC cell lines(Human colorectal cancer cell LoVo,Human colorectal cancer cell HCT116).Cell counting kit-8(CCK-8)test and flow cytometry were used to investigate the effect of overexpression of circTADA2A on the proliferation and apoptosis of LoVo and HCT116 cells.In vivo,a total of 12 nude mice were randomly divided into 2 groups using the random number table method,and HCT116 cells,transfected with circTADA2A(overexpression group)or empty vector(control group),were injected subcutaneously respectively,and the volume and weight of subcutaneous tumors between the two groups were compared.Qualitative data were analyzed by Chi-square test or Fisher′s exact test.T-test or nonparametric test were used for quantitative data.Results The relative expression of circTADA2A in CRC tissues(1.12±0.45)was significantly lower than that in adjacent tissues(0.48±0.22,t=9.620,P<0.01),and its expression was significantly correlated with tumor T stage(χ^(2)=6.855,P<0.05),lymph node metastasis(χ^(2)=10.952,P<0.05),TNM stage(χ^(2)=6.023,P<0.05)and tumor differentiation(χ^(2)=5.376,P<0.05).The results of CCK-8 test showed that the proliferation ability of LoVo and HCT116 cells in the overexpression group was significantly lower at 48 h and 72 h than in the control group:at 48 h for the overexpression group[LoVo:(0.34±0.02);HCT116:(0.31±0.03)]vs.control group[LoVo:(0.64±0.03);HCT116:(0.53±0.04),tLoVo=11.190,tHCT116=7.510,all P<0.01];

关 键 词：环状RNA 结直肠癌 增殖 脱噬作用 

分 类 号：R735.34[医药卫生—肿瘤]