宫颈癌组织及细胞株中长链非编码RNA CCAT1的表达水平及意义  

Expression level and significance of long-chain non-coding RNA CCAT1 in cervical cancer tissues and cell lines

作　　者：王圣坦[1] 朱根海[1] 纪武[1] 洪澜[1] 王雁[1] 陈春英[1] 王康[1] WANG Sheng-Tan;ZHU Gen-Hai;JI Wu;HONG Lan;WANG Yan;CHEN Chun-Ying;WANG Kang(Department of Gynecology,Hainan Provincial People′s Hospital,Haikou 570311,China)

机构地区：[1]海南省人民医院妇科,海口570311

出　　处：《中国免疫学杂志》2020年第5期583-587,593,共6页Chinese Journal of Immunology

基　　金：海南省科学技术研究与发展计划项目(817316)。

摘　　要：目的:研究宫颈癌组织及细胞株中长链非编码RNA肠癌相关转录因子1(CCAT1)的表达水平及意义。方法:采用实时荧光定量PCR(qRT-PCR)检测2015年1月至2017年6月接受宫颈癌根治术的48例宫颈癌患者的癌组织、对应癌旁正常组织(距癌<3 cm)以及人正常宫颈鳞状细胞系ECT1/E6E7、宫颈癌细胞系Hela、C33A中CCAT1的表达,并分析其临床意义;应用小分子RNA(siRNA)分别转染人宫颈癌细胞系Hela、C33A,构建CCAT1低表达细胞系,采用细胞计数试剂盒(CCK-8)法检测CCAT1对人宫颈癌细胞增殖的影响,流式细胞仪检测各组细胞周期,蛋白印迹(Western blot)法检测CCAT1对人宫颈癌细胞中RAS相关区域家族1A蛋白(RASSF1A)、细胞周期蛋白D1(Cyclin D1)表达的影响。结果:与癌旁正常组织相比,人宫颈癌组织中CCAT1的表达显著升高(P<0.05);与正常宫颈细胞ECT1/E6E7相比,宫颈癌细胞系Hela、C33A中CCAT1的表达显著升高(P<0.05);TMN分期较晚、肿瘤大、有淋巴结转移的宫颈癌患者宫颈癌组织中CCAT1相对表达量更高(P<0.05);不同年龄、分化程度患者中CCAT1的表达无显著性差异(P>0.05);与siRNA-NC组相比,转染siRNA-CCAT1的Hela、C33A细胞中CCAT1、细胞增殖活性、cyclin D1蛋白的表达显著降低,RASSF1A蛋白的表达显著升高。结论:CCAT1的表达上调与宫颈癌发生发展密切相关,沉默CCAT1的表达会抑制宫颈癌细胞的增殖,可能与上调RASSF1A的表达、下调cyclin D1的表达有关。Objective:To study the expression level and significance of long-chain non-coding RNA colorectal cancer associated transcription factor 1(CCAT1)in cervical cancer tissues and cell lines.Methods:Real-time quantitative PCR(qRT-PCR)was performed to detect expression of CCAT1 in cancer tissues,corresponding adjacent normal tissues(less than 3 cm away from cancer),human normal squamous cell line ECT1/E6E7,cervical cancer cell lines Hela and C33A of the 48 cervical cancer patients who underwent radical hysterectomy from January 2015 to June 2017.And its clinical significance was analyzed.The small interfering RNA(siRNA)was performed to transfect human cervical cancer cell lines Hela and C33A,respectively.CCAT1 low expression cell lines were constructed.Cell Counting Kit-8(CCK-8)method was performed to detect effects of CCAT1 on proliferation of human cervical cancer cells.Flow cytometry was performed to detect cell cycle in each group.Western blot was performed to detect effects of CCAT1 on expression of RAS correlation region family 1A(RASSF1A)and cyclin D1(Cyclin D1)in human cervical cancer cells.Results:Compared with adjacent normal tissues,the expression of CCAT1 in human cervical cancer tissues was significantly increased(P<0.05).Compared with normal cervical cells ECT1/E6E7,the expression of CCAT1 in cervical cancer cell lines Hela and C33A was significantly increased(P<0.05).The relative expression quantity of CCAT1 of cervical cancer tissues in cervical cancer patients with late TMN stage,large tumor and lymph node metastasis was higher(P<0.05).There was no significant difference in expression of CCAT1 among patients with different age and differentiation degree(P>0.05).Compared with siRNA-NC group,CCAT1,cell proliferation activity and expression of cyclin D1 protein were significantly decreased in Hela and C33A cells transfected with siRNA-CCAT1,while expression of RASSF1A protein was significantly increased.Conclusion:Up-regulation of CCAT1 expression is closely related to occurrence and development of cervic

关 键 词：宫颈癌组织 HELA C33A 长链非编码RNA 肠癌相关转录因子1 

分 类 号：R737.33[医药卫生—肿瘤]