摘要
目的研究环状RNA浆细胞瘤变体易位1(circPVT1)在胆囊癌中的表达水平及其与临床病理特征的关系,并分析其对胆囊癌细胞增殖和转移的影响。方法采用实时荧光定量PCR检测circPVT1在胆囊癌和癌旁组织中的表达水平,分析circPVT1的表达水平与胆囊癌患者临床病理参数的关系;常规培养胆囊癌细胞系GBC-SD,分为对照组si-NC和实验组si-circPVT1,分别转染NC siRNA和circPVT1 siRNA,实时荧光定量PCR检测circPVT1在各组细胞中的表达水平;MTt实验检测各组细胞的增殖能力;Transwell实验检测各组细胞的转移能力;裸鼠成瘤实验检测各组细胞体内成瘤能力;Western blotting检测各组细胞中Wnt/β-catenin信号通路关键蛋白Wnt5a和β-catenin的表达。结果实时荧光定量PCR结果显示,circPVT1在胆囊癌中的表达显著高于癌旁组织(P<0.05),且circPVT1的表达与患者远处转移和TNM分期相关(P<0.05)。与对照组si-NC相比,实验组si-circPVT1细胞中circPVT1的表达降低(P<0.05),si-circPVT1组细胞增殖、转移和体内成瘤能力均降低(P<0.05);与对照组si-NC相比,si-circPVT1组细胞中Wnt5a和β-catenin蛋白的表达减少(P<0.05)。结论circPVT1在胆囊癌组织中异常高表达,且其表达水平与远处转移和TNM分期相关,其可能通过调控Wnt/β-catenin信号通路促进胆囊癌的增殖和转移能力。
Objective To investigate the expression level of circular RNA plasmacytoma variant translocation 1(circPVT1)in gallbladder cancer and its relationship with clinicopathological features,and to analyze its effect on the proliferation and metastasis of gallbladder cancer cells.Methods Real-time fluorescence quantitative PCR was conducted to detect the expression level of circPVT1 in gallbladder cancer and adjacent tissues.Gallbladder carcinoma cell line GBC-SD were routine cultured,and cells were divided into the control group(si-NC)and experimental group(si-circPVT1).NC siRNA and circPVT1 siRNA were transfected respectively,and the expression level of circPVT1 in each group of cells was detected by real-time fluorescence quantitative PCR.MTt experiment was performed to detect the proliferation ability of each group of cells.Transwell method was conducted to detect the metastasis ability of each group of cells.Nude mouse tumor formation experiment was applied to detect the tumorigenic ability of each group of cells in vivo.Western blotting was used to detect the expression of the key protein Wnt5a andβ-catenin of Wnt/β-catenin signaling pathway in each group of cells.Results The results of real-time fluorescence quantitative PCR showed that the expression of circPVT1 in gallbladder cancer was significantly higher than that in adjacent tissues(P<0.05),and the expression of circPVT1 was correlated with distant metastasis and TNM staging(P<0.05).Compared with the control group(si-NC),the expression of circPVT1 in cells of the experimental group(si-circPVT1)was reduced(P<0.05),and the cell proliferation,metastasis and tumor formation ability in the si-circPVT1 group were reduced(P<0.05).Compared with the control group(si-NC),the expression of Wnt5a andβ-catenin protein in cells of the si-circPVT1 group were reduced(P<0.05).Conclusion CircPVT1 is highly expressed in gallbladder carcinoma tissues,and its expression level is related to distant metastasis and TNM stage.It may promote the proliferation and metastasis of gallbladder carcinoma by regulating Wnt/β-catenin signaling pathway.
作者
杨丹琪
周影
马慧
YANG Danqi;ZHOU Ying;MA Hui(Department of Geriatrics, Xiamen hospital, Zhongshan Hospital Affiliated to Fudan University, Xiamen 361006, China;Endoscopy Center of Xiamen Hospital, Zhongshan Hospital Affiliated to Fudan University, Xiamen 361006, China;Department of Geriatrics, Zhongshan Hospital Affiliated to Fudan University, Shanghai 200032, China)
出处
《标记免疫分析与临床》
CAS
2021年第6期922-926,931,共6页
Labeled Immunoassays and Clinical Medicine
基金
厦门市医疗卫生指导性项目(编号:3502Z20199178)。
