Establishment and characterization of a novel neuroendocrine carcinoma cell line derived from a human ascending colon tumor

Abstract

The incidence of rare neuroendocrine tumors (NET) is rapidly increasing. Neuroendocrine carcinoma (NEC) is a NET with poorly differentiated histological features, high proliferative properties and associated poor prognoses. As these carcinomas are so rare and, thus, affect only a small number of patients allowing for few cell lines to be derived from patient biopsies, the histological, immunohistochemical, and clinical characteristics associated with colorectal NEC and NEC in other organs have yet to be clearly defined. Herein, we describe the establishment of a novel NEC cell line (SS‐2) derived from a tumor resection of the ascending colon from a 59‐year‐old Japanese woman. The histological, electron microscopic and immunohistochemical features of chromogranin A (CgA) as well as confirmation of synaptophysin positivity in this tumor were typical of those commonly observed in surgically resected colorectal NEC. Further, the Ki‐67 labeling index of the resected tumor was >20% and, thus, the tumor was diagnosed as an NEC of the ascending colon. The SS‐2 cell line maintained characteristic features to those of the resected tumor, which were further retained following implantation into subcutaneous tissues of nude mice. Additionally, when SS‐2 cells were seeded into ultra‐low attachment plates, they formed spheres that expressed higher levels of the cancer stem cell (CSC) marker CD133 compared to SS‐2 cells cultured under adherent conditions. SS‐2 cells may, therefore, contribute to the current knowledge on midgut NEC biological function while providing a novel platform for examining the effects of colorectal NEC drugs, including CSC.

Extrapulmonary poorly differentiated NECs, including molecular and immune aspects

“…Shinji et al recently established a novel cell line (SS-2) derived from resection of an ascending colon tumour (Shinji et al 2019). The SS-2 cell line maintained characteristic features of the resected tumour, which were also retained when implanted into the s.c. tissue of nude mice, and the authors concluded that these cells may add to the current knowledge of the biological behaviour of midgut NEC and thus may provide a novel therapeutic examination platform (Shinji et al 2019).…”

Isolation and Characterization of Two Novel Colorectal Cancer Cell Lines, Containing a Subpopulation with Potential Stem-Like Properties: Treatment Options by MYC/NMYC Inhibition

“…CD133 is a robust biomarker to identify primary CSC and can be proposed as a prognostic marker of CRC patients. Moreover, it is reported to be expressed in poorly-differentiated NECs and well-differentiated NETs of the digestive tract [ 34 ] and was demonstrated to be expressed in CCSC-lines [ 35 ]. It is the most well-known marker for the isolation and investigation of CSC in different types of cancer [ 36 , 37 , 38 ] and known to be responsible for radio- and chemotherapy-resistance in CCSC [ 39 , 40 ].…”

From Donor to the Lab: A Fascinating Journey of Primary Cell Lines

“…In ascending colon cancer, WB can be useful for the determination of the presence and size of insulinoma-associated protein 1 (INSM-1) from lysates. INSM-1 is a product of gene INSM-1 that regulates the expression of neuroendocrine markers (Shinji et al, 2019).…”

“…Moreover, RT-qPCR is commonly used as quality control to examine changes in expression during RNA sequencing (RNA-seq) (Adams, 2020). In the characterisation of cell lines, RT-qPCR can be used to investigate cancer stem cell markers, such as expression of clusters of differentiation (CD24, CD44, CD133, and CD166) (Shinji et al, 2019). This method also enables the detection of particular genes needed for cell line characterisation, for example, B2M and RPL29, in the case of tongue carcinoma cell line (Wang et al, 2017).…”