Jing S, Feng Y, He X L, Wang Y
Department of Gynaecology, Henan Province People's Hospital, People's Hospital of Zhengzhou University, People's Hospital of Henan University, Zhengzhou 450003, China.
Zhonghua Zhong Liu Za Zhi. 2023 Jan 23;45(1):56-63. doi: 10.3760/cma.j.cn112152-20200623-00588.
To investigate the effect of long non-coding RNA urothelial carcinoma-associated 1 (UCA1) gene on the proliferation, migration, apoptosis and immune escape of endometrial cancer cells and its molecular mechanism. Endometrial cancer tissues and adjacent normal tissues of patients with endometrioid adenocarcinoma who underwent total or partial hysterectomy in Henan Provincial People's Hospital from 2017 to 2019 were collected. The expressions of UCA1 and miR-204-5p were detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR), and the cell proliferation, migration and apoptosis were detected by cell counting kit 8 (CCK8) method, Transwell method, flow cytometry, and dual-luciferase reporter assay was used to explore the target relationship between UCA1 and miR-204-5p. HEC-1A-sh-NC or HEC-1A-sh-UCA1 cells were co-cultured with peripheral blood mononuclear cells (PBMC) or cytokine-induced killer cells in vitro to explore the role of UCA1 in immune escape. The expression level of UCA1 in endometrial cancer tissue (17.08±0.84) was higher than that in adjacent normal endometrial tissue (3.00±0.37), and the expression level of miR-204-5p (0.98±0.16) was lower than that in adjacent normal endometrial tissue (2.00±0.20, <0.05). Pearson correlation analysis showed that the expression of miR-204-5p was negatively correlated with the expression of UCA1 (=-0.330, =0.030). The expressions of UCA1 and miR-204-5p were associated with the International Federation of Gynecology and Obstetrics stage of endometrial cancer, lymph node metastasis and vascular invasion (<0.05). The relative ratio of absorbance (0.58±0.11) and the number of cell migration [(199.68±18.44)] in the sh-UCA1 group were lower than those in the sh-NC group (1.24±0.17 and 374.76±24.83), respectively. The apoptosis rate of sh-UCA1 group [(28.64±7.80)%] was higher than that of sh-NC group [(14.27±4.38)%, <0.05]. After different ratios of effector cells and target cells were cultured, the cell survival rate of HEC-1A-sh-UCA1 group was lower than that of HEC-1A-sh-NC group, and the difference was statistically significant (<0.05). UCA1 had a binding site for miR-204-5p. The relative ratio of absorbance (1.74±0.08) and the number of cell migration (426.00±18.00) cells in the UCA1+ anti-miR-204-5p group were higher than those in the control group [1.00±0.03 and (284.00±8.00) cells, respectively]. The apoptosis rate of UCA1+ anti-miR-204-5p group [(5.42±0.93)%] was lower than that of control group [(14.82±1.48)%, <0.05]. HEC-1A-sh-UCA1 cells could induce higher interferon gamma (IFN-γ) expression when co-cultured with PBMC, and the levels of IFN-γ expression in PHA group and PHA+ pre-miR-204-5p group cells were 2.42±0.49 and 1.88±0.26, which were higher than that in the PHA+ pre-NC group (0.85±0.10, <0.05). When co-cultured with cytokine-induced killer cells (different ratios) in vitro, the HEC-1A-sh-UCA1 group and the HEC-1A-pre-miR-204-5p group had lower survival rates than that in the HEC-1A-pre-miR-204-5p group. In the HEC-1A-pre-NC group, the differences were statistically significant (<0.05). UCA1/miR-204-5p may play an important role in human endometrial cancer.
探讨长链非编码RNA尿路上皮癌相关1(UCA1)基因对子宫内膜癌细胞增殖、迁移、凋亡及免疫逃逸的影响及其分子机制。收集2017年至2019年在河南省人民医院接受全子宫或部分子宫切除术的子宫内膜样腺癌患者的子宫内膜癌组织及癌旁正常组织。采用实时荧光定量聚合酶链反应(RT-qPCR)检测UCA1和miR-204-5p的表达,采用细胞计数试剂盒8(CCK8)法、Transwell法、流式细胞术检测细胞增殖、迁移和凋亡,并采用双荧光素酶报告基因检测法探讨UCA1与miR-204-5p的靶向关系。将HEC-1A-sh-NC或HEC-1A-sh-UCA1细胞与外周血单个核细胞(PBMC)或细胞因子诱导的杀伤细胞体外共培养,探讨UCA1在免疫逃逸中的作用。子宫内膜癌组织中UCA1的表达水平(17.08±0.84)高于癌旁正常子宫内膜组织(3.00±0.37),miR-204-5p的表达水平(0.98±0.16)低于癌旁正常子宫内膜组织(2.00±0.20,P<0.05)。Pearson相关性分析显示,miR-204-5p的表达与UCA1的表达呈负相关(r=-0.330,P=0.030)。UCA1和miR-204-5p的表达与子宫内膜癌的国际妇产科联盟分期、淋巴结转移及血管浸润相关(P<0.05)。sh-UCA1组的吸光度相对比值(0.58±0.11)和细胞迁移数[(199.68±18.44)]均低于sh-NC组(1.24±0.17和374.76±24.83)。sh-UCA1组的凋亡率[(28.64±7.80)%]高于sh-NC组[(14.27±4.38)%,P<0.05]。不同效应细胞与靶细胞比例培养后,HEC-1A-sh-UCA1组的细胞存活率低于HEC-1A-sh-NC组,差异有统计学意义(P<0.05)。UCA1有一个与miR-204-5p的结合位点。UCA1+抗miR-204-5p组的吸光度相对比值(1.74±0.08)和细胞迁移数(426.00±18.00)均高于对照组[分别为1.00±0.03和(284.00±8.00)]。UCA1+抗miR-204-5p组的凋亡率[(5.42±0.93)%]低于对照组[(14.82±1.48)%,P<0.05]。HEC-1A-sh-UCA1细胞与PBMC共培养时可诱导更高水平的干扰素γ(IFN-γ)表达,PHA组和PHA+pre-miR-204-5p组细胞中IFN-γ的表达水平分别为2.42±0.49和1.88±0.26,高于PHA+pre-NC组(0.85±0.10,P<0.05)。体外与细胞因子诱导的杀伤细胞(不同比例)共培养时,HEC-1A-sh-UCA1组和HEC-1A-pre-miR-204-5p组的存活率低于HEC-1A-pre-NC组,差异有统计学意义(P<0.05)。UCA1/miR-204-5p可能在人类子宫内膜癌中起重要作用。