Frontiers in Oncology (Oct 2022)
Establishment and characterization of an immortalized epithelial cell line from human gallbladder
- Ziyi Wang,
- Ziyi Wang,
- Ziyi Wang,
- Ziyi Wang,
- Shijia Wang,
- Shijia Wang,
- Shijia Wang,
- Shijia Wang,
- Ziheng Jia,
- Ziheng Jia,
- Ziheng Jia,
- Ziheng Jia,
- Yuhao Zhao,
- Yuhao Zhao,
- Yuhao Zhao,
- Yuhao Zhao,
- Mao Yang,
- Mao Yang,
- Mao Yang,
- Mao Yang,
- Weikang Yan,
- Weikang Yan,
- Weikang Yan,
- Weikang Yan,
- Tao Chen,
- Tao Chen,
- Tao Chen,
- Tao Chen,
- Dongxi Xiang,
- Dongxi Xiang,
- Dongxi Xiang,
- Rong Shao,
- Rong Shao,
- Rong Shao,
- Rong Shao,
- Rong Shao,
- Yingbin Liu,
- Yingbin Liu,
- Yingbin Liu,
- Yingbin Liu
Affiliations
- Ziyi Wang
- Department of Biliary-Pancreatic Surgery, Renji Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China
- Ziyi Wang
- Shanghai Cancer Institute, State Key Laboratory of Oncogenes and Related Genes, Shanghai, China
- Ziyi Wang
- Shanghai Key Laboratory of Biliary Tract Disease, Renji Hospital, Shanghai, China
- Ziyi Wang
- Shanghai Research Center of Biliary Tract Disease, Renji Hospital, Shanghai, China
- Shijia Wang
- Department of Biliary-Pancreatic Surgery, Renji Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China
- Shijia Wang
- Shanghai Cancer Institute, State Key Laboratory of Oncogenes and Related Genes, Shanghai, China
- Shijia Wang
- Shanghai Key Laboratory of Biliary Tract Disease, Renji Hospital, Shanghai, China
- Shijia Wang
- Shanghai Research Center of Biliary Tract Disease, Renji Hospital, Shanghai, China
- Ziheng Jia
- Department of Biliary-Pancreatic Surgery, Renji Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China
- Ziheng Jia
- Shanghai Cancer Institute, State Key Laboratory of Oncogenes and Related Genes, Shanghai, China
- Ziheng Jia
- Shanghai Key Laboratory of Biliary Tract Disease, Renji Hospital, Shanghai, China
- Ziheng Jia
- Shanghai Research Center of Biliary Tract Disease, Renji Hospital, Shanghai, China
- Yuhao Zhao
- Department of Biliary-Pancreatic Surgery, Renji Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China
- Yuhao Zhao
- Shanghai Cancer Institute, State Key Laboratory of Oncogenes and Related Genes, Shanghai, China
- Yuhao Zhao
- Shanghai Key Laboratory of Biliary Tract Disease, Renji Hospital, Shanghai, China
- Yuhao Zhao
- Shanghai Research Center of Biliary Tract Disease, Renji Hospital, Shanghai, China
- Mao Yang
- Department of Biliary-Pancreatic Surgery, Renji Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China
- Mao Yang
- Shanghai Cancer Institute, State Key Laboratory of Oncogenes and Related Genes, Shanghai, China
- Mao Yang
- Shanghai Key Laboratory of Biliary Tract Disease, Renji Hospital, Shanghai, China
- Mao Yang
- Shanghai Research Center of Biliary Tract Disease, Renji Hospital, Shanghai, China
- Weikang Yan
- Department of Biliary-Pancreatic Surgery, Renji Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China
- Weikang Yan
- Shanghai Cancer Institute, State Key Laboratory of Oncogenes and Related Genes, Shanghai, China
- Weikang Yan
- Shanghai Key Laboratory of Biliary Tract Disease, Renji Hospital, Shanghai, China
- Weikang Yan
- Shanghai Research Center of Biliary Tract Disease, Renji Hospital, Shanghai, China
- Tao Chen
- Department of Biliary-Pancreatic Surgery, Renji Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China
- Tao Chen
- Shanghai Cancer Institute, State Key Laboratory of Oncogenes and Related Genes, Shanghai, China
- Tao Chen
- Shanghai Key Laboratory of Biliary Tract Disease, Renji Hospital, Shanghai, China
- Tao Chen
- Shanghai Research Center of Biliary Tract Disease, Renji Hospital, Shanghai, China
- Dongxi Xiang
- Shanghai Cancer Institute, State Key Laboratory of Oncogenes and Related Genes, Shanghai, China
- Dongxi Xiang
- Shanghai Key Laboratory of Biliary Tract Disease, Renji Hospital, Shanghai, China
- Dongxi Xiang
- Shanghai Research Center of Biliary Tract Disease, Renji Hospital, Shanghai, China
- Rong Shao
- Department of Biliary-Pancreatic Surgery, Renji Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China
- Rong Shao
- Shanghai Cancer Institute, State Key Laboratory of Oncogenes and Related Genes, Shanghai, China
- Rong Shao
- Shanghai Key Laboratory of Biliary Tract Disease, Renji Hospital, Shanghai, China
- Rong Shao
- Shanghai Research Center of Biliary Tract Disease, Renji Hospital, Shanghai, China
- Rong Shao
- Department of Pharmacology and Biochemistry, Shanghai Jiao Tong University School of Medicine, Shanghai, China
- Yingbin Liu
- Department of Biliary-Pancreatic Surgery, Renji Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China
- Yingbin Liu
- Shanghai Cancer Institute, State Key Laboratory of Oncogenes and Related Genes, Shanghai, China
- Yingbin Liu
- Shanghai Key Laboratory of Biliary Tract Disease, Renji Hospital, Shanghai, China
- Yingbin Liu
- Shanghai Research Center of Biliary Tract Disease, Renji Hospital, Shanghai, China
- DOI
- https://doi.org/10.3389/fonc.2022.994087
- Journal volume & issue
-
Vol. 12
Abstract
BackgroundAlthough a plethora of studies have employed multiple gallbladder cancer (GBC) cell lines, it is surprisingly noted that there is still lack of a normal gallbladder epithelial cell line as a normal counterpart, thus impeding substantially the progress of mechanistic studies on the transformation of normal epithelial cells to cancer. Here, we created a normal gallbladder epithelial cell line named L-2F7 from human gallbladder tissue.MethodsGallbladder tissues from a diagnosed cholecystitis female patient were collected, and epithelial cells were enriched by magnetic cell sorting. Then, the cells were immortalized by co-introduction of human telomerase reverse transcriptase (hTERT) and Simian virus 40 large T antigen (LT-SV40) via a lentivirus infection system. After clonal selection and isolation, L-2F7 cells were tested for epithelial markers CK7, CK19, CK20, and CD326, genomic feature, cell proliferation, and migration using Western blot, immunofluorescence, whole genome sequencing, karyotyping, and RNA sequencing. L-2F7 cells were also transplanted to Nude (nu/nu) mice to determine tumorigenicity.ResultsWe successfully identified one single-cell clone named L-2F7 which highly expressed epithelial markers CD326, CK7, CK19, and CK20. This cell line proliferated with a doubling time of 23 h and the epithelial morphology sustained over 30 passages following immortalization. Transient gene transduction of L-2F7 cells led to expression of exogenous GFP and FLAG protein. L-2F7 cells exhibited both distinct non-synonymous mutations from those of gallbladder cancer tissues and differential non-cancerous gene expression patterns similar to normal tissue. Although they displayed unexpected mobility, L-2F7 cells still lacked the ability to develop tumors.ConclusionWe developed a non-cancerous gallbladder epithelial cell line, offering a valuable system for the study of gallbladder cancer and other gallbladder-related disorders.
Keywords
