摘要
目的:探讨CCT2(chaperonin containning TCP1,subunit 2)在人肺腺癌中的表达、对肺腺癌细胞增殖、迁移生物学功能的影响及其与患者临床特征的关系。方法:通过TCGA和GTEx数据库分析在肺腺癌组织及正常肺组织样本中CCT2的表达差异,生存分析评价其表达水平与患者预后的关系。免疫组织化学法分别检测CCT2、Ki-67在72例肺腺癌组织中的表达水平,并分析其表达水平与临床病理特征之间的相关性。采用逆转录-聚合酶链反应(qRT-PCR)、Western blot检测CCT2在肺腺癌细胞中的表达情况;A549、H1650、H1299细胞株分别转染siRNA,分为si-NC组和si-CCT2组,qRT-PCR、Western blot验证转染效率,CCK-8实验检测肺腺癌细胞增殖能力,细胞划痕实验检测细胞迁移能力。结果:基于TCGA数据库的肺腺癌数据分析表明,与正常肺组织相比,肺腺癌组织中CCT2的表达水平更高,CCT2的高表达与肺腺癌患者的总体生存率降低相关(P<0.05)。qRT-PCR、Western blot检测显示,CCT2在肺腺癌细胞中高表达,免疫组化显示CCT2的表达与Ki-67表达呈正相关(P<0.05)。CCK-8实验检测结果显示,A549、H1650细胞在转染siRNA 48 h后,相比si-NC组,si-CCT2组细胞增殖能力降低,差异有统计学意义(P<0.05)。细胞划痕实验结果显示,A549、H1650细胞在转染siRNA 24 h后,si-CCT2组细胞的迁移率低于si-NC组,差异均有统计学意义(P<0.05)。结论:CCT2在肺腺癌细胞和组织中表达水平增加,敲低其表达可减弱A549、H1650肺腺癌细胞的增殖及迁移。
Objective:The current study aims to explore the biological function of CCT2(chaperonin containning TCP1,subunit 2)in human lung adenocarcinoma.Methods:The expression divergence of CCT2 in lung adenocarcinoma tissues and normal lung tissue samples were analyzed from TCGA and GTEx database.The relationship between the expression level of CCT2 and the patient’s prognosis was evaluated through survival analysis.The expression of CCT2 in lung adenocarcinoma cells was measured by Reverse Transcription-Polymerase Chain Reaction(qRT-PCR)and Western blot.CCT2 siRNA was transfected into cell strains(A549,H1650 or H1299)respectively,which were divided into Group si-NC(negative control group)and Group si-CCT2(transfection group).Transfection efficiency was detected by qRT-PCR and Western blot.The proliferation ability of lung adenocarcinoma cell was tested by CCK-8.The cellular migration ability was detected by wound-healing experiment.The expression levels of CCT2 and Ki-67 in 72 cases of lung adenocarcinoma tissues were detected with immunohistochemical method,and the correlation between their expression levels and clinical pathology features were analyzed.Results:The results showed that,compared with normal lung tissues,the expression level of CCT2 in lung adenocarcinoma tissue was higher,which was related to low overall survival rate of lung adenocarcinoma patients(P<0.05).According to qPCR and Western blot test,CCT2 highly expressed in lung adenocarcinoma cell,and the immunohistochemistry result showed that the CCT2 expression was positively correlated with the Ki-67 expression(P<0.05).CCK-8 test results showed that the proliferation ability of A549 and H1650 cells in si-CCT2 group was significantly lower than that in the siNC group 48 hours after siRNA transfection(P<0.05).The results of cell scratch test showed that the migration rate of A549 and H1650 cells in the si-CCT2 group was lower than that in the si-NC group 24 hours after siRNA transfection(P<0.05).Conclusion:The expression levels of CCT2 in lung adenocarcinoma cells and tissues increases.Knocking down its expression can inhibit the proliferation and migration of A549 and H1650 lung adenocarcinoma cells.
作者
何树燕
沈冬
林青凤
陈洁
马成龙
HE Shuyan;SHEN Dong;LIN Qingfeng;CHEN Jie;MA Chenglong(Department of Oncology,Jiangyin Hospital Affiliated to Nantong University,Jiangyin 214400,China)
出处
《南京医科大学学报（自然科学版）》
CAS
北大核心
2023年第3期319-325,共7页
Journal of Nanjing Medical University(Natural Sciences)
基金
江苏省卫生健康委员会面上项目(H2019103)
江阴市卫健委面上项目(S202101)
江阴市卫健委面上项目(S202003)。
关键词
肺腺癌
CCT2
增殖
迁移
lung adenocarcinoma
CCT2
proliferation
migration
