摘要
目的研究肿瘤蛋白翻译调节因子1反义-RNA1(TPT1-AS1)在肝细胞和肝癌细胞中表达的差异,及其对肝细胞癌增殖、迁移、侵袭、上皮细胞间质化(EMT)的影响。方法2018年9月—2020年6月于西安交通大学实验室进行细胞实验,使用qRT-PCR分别检测92例经手术切除的肝癌组织与对应癌旁肝组织,肝癌细胞HepG2、SNU-182和人肝永生化细胞THLE-3中TPT1-AS1相对表达量。根据qRT-PCR测得肝癌组织中TPT1-AS1表达量中位数(3.38),将患者分为TPT1-AS1高表达组(n=46)和低表达组(n=46)。使用Kaplan-Meier法分析比较2组患者5年生存率的差异。使用siRNA转染HepG2和SNU-182细胞以沉默TPT1-AS1表达,以阴性对照siRNA(si-NC)转染相同细胞作为对照。比较siRNA和si-NC转染HepG2和SNU-182细胞后的增殖、迁移、侵袭、EMT能力差异。结果与对应癌旁组织比较,肝癌组织中TPT1-AS1相对表达水平升高(t/P=54.065/0.000)。高表达组患者BCLC分期C期比例、淋巴结转移比例更高(χ^(2)/P=15.187/0.001,7.379/0.007)。高表达组患者5年生存率低于低表达组(χ^(2)/P=7.354/0.007)。使用siRNA转染的HepG2和SNU-182细胞较si-NC转染对应细胞的细胞集落数量明显减少,G0/G1期细胞比例升高(P均=0.000),S期细胞比例显著降低(P均=0.000),相对迁移距离降低(t/P=25.153/0.000),穿透基底膜的细胞数量减少,G1/S期转换及EMT相关蛋白CDK4、N-cadherin和Vimentin的相对表达量降低(P均=0.000),而p21和E-cadherin蛋白相对表达量升高(P均=0.000)。结论长链非编码RNA TPT1-AS1促进肝癌细胞增殖、迁移、侵袭和EMT,TPT1-AS1有可能作为肝细胞癌的治疗靶点。
Objective To study the difference in expression of tumor protein translation regulator 1 antisense RNA1(TPT1-AS1)in hepatocytes and hepatocarcinoma cells,and its effects on hepatocellular carcinoma proliferation,migration,invasion,and epithelial cell interstitialization(EMT).Methods From September 2018 to June 2020,cell experiments were carried out in the Laboratory of Xi'an Jiaotong University,using qRT-PCR to detect 92 cases of surgically resected liver cancer tissues and corresponding adjacent liver tissues,liver cancer cells HepG2,SNU-182 and human liver immortalized cells The relative expression of TPT1-AS1 in THLE-3.According to the median of TPT1-AS1 expression in liver cancer tissue measured by qRT-PCR(3.38),patients were divided into TPT1-AS1 high expression group(n=46)and low expression group(n=46).Kaplan-Meier method was used to analyze and compare the difference in 5-year survival rate between the two groups.HepG2 and SNU-182 cells were transfected with siRNA to silence TPT1-AS1 expression,and the same cells were transfected with negative control siRNA(si-NC)as a control.Compare the proliferation,migration,invasion and EMT ability of siRNA and si-NC transfected HepG2 and SNU-182 cells.Results Compared with the corresponding adjacent tissues,the relative expression level of TPT1-AS1 in liver cancer tissues increased(t/P=54.065/0.000).The proportion of BCLC stage C and lymph node metastasis were higher in the high expression group(χ^(2)/P=15.187/0.001,7.379/0.007).The 5-year survival rate of patients in the high expression group was lower than that in the low expression group(χ^(2)/P=7.354/0.007).Compared with the corresponding cells transfected with si NC,the number of cell colonies in HepG2 and SNU-182 cells transfected with siRNA was significantly reduced,the ratio of cells in G0/G1 phase was increased(P=0.000),and the ratio of cells in S phase was significantly reduced(P=0.000),The relative migration distance is reduced(t/P=25.153/0.000),the number of cells penetrating the basement membrane is reduced,the G1/S phase transition and the relative expression of EMT-related proteins CDK4,N-cadherin and Vimentin are reduced(P=0.000),While the relative expression of p21 and E cadherin protein increased(P=0.000).Conclusion The long non-coding RNA TPT1-AS1 promotes the proliferation,migration,invasion and EMT of liver cancer cells.TPT1-AS1 may be used as a therapeutic target for hepatocellular carcinoma.
作者
王斐斐
何婷
李炜
孙琳
曲凯
姚忠强
Wang Feifei;He Ting;Li Wei;Sun Lin;Qu Kai;Yao Zhongqiang(Department of Oncology,No.3201 Affiliated Hospital of Xi'an Jiaotong University,Shaanxi Province,Hanzhong 723000,China;不详)
出处
《疑难病杂志》
CAS
2022年第1期24-30,共7页
Chinese Journal of Difficult and Complicated Cases
基金
陕西省自然科学基础计划研究项目(2017JM8039)。
