Zhou Fei, Zhang Yanhua, Sun Jihong, Yang Xiaoming
1Department of Radiology, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang China.
2Department of Pathology, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang China.
Cancer Cell Int. 2019 Jul 22;19:190. doi: 10.1186/s12935-019-0911-1. eCollection 2019.
Gallbladder cancer is the most common malignant neoplasm of the biliary tract, responsible for 80-95% of cases. Appropriate models are required for investigating the molecular pathogenesis of gallbladder cancer.
In this study, we aimed to establish a gallbladder cancer cell line from primary tumour. Single cell RNA sequencing, Light and electron microscopy, DNA content analysis, cytogenetic analysis, short tandem repeat (STR) DNA fingerprint analysis, immunophenotypic characterization, and xeno-transplantation were utilized to characterize the novel ZJU-0430 cell line in vitro and in vivo.
The cell line showed multiple cell shapes and characteristic epithelial morphologies under the microscope, but no too much heterogeneity by scRNA-Seq, with a population doubling time (PDT) of 19.81 h, which was shorter than that for GBC-SD cells. An immunophenotypic analysis revealed that ZJU-0430 cells were positive for CD24, CD44, CD29 and CD133 expression, and partially positive for CD184, and CD326 expression, and negative for CD34, CD90, CD117, and CD338 expression, similar to the primary cancer cells. A pathological analysis confirmed the origination of cell line from gallbladder tumour. ZJU-0430 cells had higher migration, invasion and proliferation properties than GBC-SD cells in vitro, and showed in vivo tumorigenicity in nude mouse xenograft settings.
The results confirm the potential utility of ZJU-0430 cell line as a representative model of gallbladder cancer and suggest that it could be used in the in vitro and in vivo studies of gallbladder cancer pathogenesis and to develop new therapeutics.
胆囊癌是胆道最常见的恶性肿瘤，占病例的80 - 95%。研究胆囊癌的分子发病机制需要合适的模型。
在本研究中，我们旨在从原发性肿瘤建立胆囊癌细胞系。利用单细胞RNA测序、光镜和电镜、DNA含量分析、细胞遗传学分析、短串联重复序列（STR）DNA指纹分析、免疫表型特征分析和异种移植，在体外和体内对新的ZJU - 0430细胞系进行表征。
该细胞系在显微镜下呈现多种细胞形态和典型的上皮形态，但通过单细胞RNA测序显示异质性不大，群体倍增时间（PDT）为19.81小时，短于GBC - SD细胞。免疫表型分析显示，ZJU - 0430细胞CD24、CD44、CD29和CD133表达呈阳性，CD184和CD326表达部分呈阳性，CD34、CD90、CD117和CD338表达呈阴性，与原发性癌细胞相似。病理分析证实该细胞系起源于胆囊肿瘤。ZJU - 0430细胞在体外比GBC - SD细胞具有更高的迁移、侵袭和增殖特性，并且在裸鼠异种移植模型中显示出体内致瘤性。
结果证实ZJU - 0430细胞系作为胆囊癌代表性模型的潜在效用，并表明它可用于胆囊癌发病机制的体外和体内研究以及开发新的治疗方法。
