Department of Oncology,Jiangyin Hospital Affiliated to Nantong University
Jiangsu Provincial Health Commission (H2019103); Jiangyin Municipal Health Commission (S202101); Jiangyin Municipal Health Commission (S202003)
目的：探讨CCT2（chaperonin containning TCP1, subunit2）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，采用逆转录-聚合酶链反应（RT-PCR）、Western blot检测CCT2在肺腺癌细胞中的表达情况；A549、H1650、H1299细胞株中分别转染siRNA，分为si-NC组和si-CCT2组，RT-PCR、Western blot验证转染效率，采用CCK8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，使用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。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）。RT-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）。CCK8实验检测结果显示，A549、H1650细胞在转染siRNA 48小时后，NC组比si-CCT2组细胞增殖能力更强，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小时后，NC组细胞迁移距离短于siRNA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，敲低其表达水平后可减弱A549、H1650肺腺癌细胞增殖及迁移。
Objective: To explore the biological function of CCT2 (chaperonin containning TCP1,subunit2) in human lung adenocarcinoma. Method: The expression date of CCT2 in lung adenocarcinoma tissue and normal lung tissue sample were from TCGA and GTEx database, the relationship between the expression level of CCT2 and the patient’s prognosis were evaluated through survival analysis, the expression of CCT2 in lung adenocarcinoma cells were measured by Reverse Transcription-Polymerase Chain Reaction (RT-PCR) and Western blot; CCT2 siRNA was transfected into cell strains (A549, H1650 or H1299) respectively, which were divided into Group NC (negative control group) and Group si-CCT2 (transfection group). RT-PCR was used to show transfection efficiency, the proliferation ability of lung adenocarcinoma cell was tested by CCK8, the cell’s migration ability was detected by wound-healing experiment, the expression level of CCT2 and Ki-67 in 72 cases of lung adenocarcinoma tissues were detected with immunohistochemical method, and the correlation between its expression level and clinical pathology features were analyzed. Result: The data 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 date showed that CCT2 expression was positively correlated with Ki-67 expression (P＜0.05). For CCK8 test, after 48 hours of transfection, A549 and H1650 cells were significant difference of absorbance value between the cells in Group NC and Group si-CCT2 (P＜0.05). In regard to the date of wound-healing experiment, after 24 hours of transfection, the migration of cells in Group NC is remarbly shorter than that in Group siRNA (P＜0.05). Conclusion: The lung adenocarcinoma cell can be induced by CCT2 for proliferation and migration, the high expression level of CCT2 may be closely concerned with the occurrence and development of lung adenocarcinoma. CCT2 could be as the index to judge the prognosis of lung adenocarcinoma.