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Generation and sequencing of pulmonary carcinoid tumor cell lines
Michael K Asiedu1, Charles F Thomas, Sandra C Tomaszek
1*Division of General Thoracic Surgery, Department of Surgery, Mayo Clinic College of Medicine, Rochester, MN; †Thoracic Diseases Research Unit, Division of Pulmonary, Critical Care, and Internal Medicine, Department of Medicine; ‡Department of Laboratory Medicine and Pathology; and §Department of Health Sciences Research, Mayo Clinic College of Medicine, Rochester, MN.
Introduction:
Pulmonary carcinoid tumors account for approximately 5% of all lung malignancies in adults, and comprise 30% of all carcinoid tumors. There are limited reagents available to study these rare tumors, and consequently no major advances have been made for patient treatment. We report the generation and characterization of human pulmonary carcinoid tumor cell lines to study underlying biology, and to provide models for testing novel chemotherapeutic agents.
Methods:
Tissue was harvested from three patients with primary pulmonary typical carcinoid tumors undergoing surgical resection. The tumor was dissociated and plated onto dishes in culture media. The established cell lines were characterized by immunohistochemistry, Western blotting, and cell proliferation assays. Tumorigenicity was confirmed by soft agar growth and the ability to form tumors in a mouse xenograft model. Exome and RNA sequencing of patient tumor samples and cell lines was performed using standard protocols.
Results:
Three typical carcinoid tumor lines grew as adherent monolayers in vitro, expressed neuroendocrine markers consistent with the primary tumor, and formed colonies in soft agar. A single cell line produced lung tumors in nude mice after intravenous injection. Exome and RNA sequencing of this cell line showed lineage relationship with the primary tumor, and demonstrated mutations in a number of genes related to neuronal differentiation.
Conclusion:
Three human pulmonary typical carcinoid tumor cell lines have been generated and characterized as a tool for studying the biology and novel treatment approaches for these rare tumors.
Insights
Researchers developed and characterized three human pulmonary carcinoid tumor cell lines. These models will aid in studying rare lung neuroendocrine tumor biology and testing new treatments.
Area of Science:
- Oncology
- Cell Biology
- Genomics
Background:
- Pulmonary carcinoid tumors are rare lung malignancies, representing 5% of adult lung cancers.
- Limited reagents hinder research and treatment advancements for these neuroendocrine tumors.
- Novel cell line models are crucial for understanding tumor biology and therapeutic development.
Purpose of the Study:
- To generate and characterize human pulmonary carcinoid tumor cell lines.
- To establish in vitro and in vivo models for studying tumor biology.
- To provide platforms for testing novel chemotherapeutic agents.
Main Methods:
- Tissue harvesting from three typical pulmonary carcinoid tumor patients.
- Establishment and characterization of cell lines using immunohistochemistry, Western blotting, and proliferation assays.
- Tumorigenicity assessment via soft agar growth and mouse xenograft models; exome and RNA sequencing performed.
Main Results:
- Three adherent pulmonary carcinoid tumor cell lines were established, expressing neuroendocrine markers.
- One cell line demonstrated tumor formation in nude mice, confirming lineage relationship via sequencing.
- Sequencing revealed mutations in genes associated with neuronal differentiation.
Conclusions:
- Successfully generated and characterized three human pulmonary carcinoid tumor cell lines.
- These cell lines serve as valuable tools for investigating the biology of rare pulmonary neuroendocrine tumors.
- The models facilitate the exploration of novel therapeutic strategies for improved patient treatment.
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