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2000
Volume 20, Issue 4
  • ISSN: 1574-8928
  • E-ISSN: 2212-3970

Abstract

Background

Colorectal adenoma represents the critical step in the development of colorectal cancer. The establishment of an immortalized epithelial cell line of colorectal adenoma of human origin would provide a tool for studying the mechanism of precancerous lesions, screening the efficacy of novel drugs, and constructing disease models. Currently, there is no commercially available stable supply of epithelial cells from precancerous lesions.

Aims

This study aimed to establish a natural LHPP low-expressing precancerous epithelial cell line by SV40-LT antigen gene transfection.

Methods

Simian vacuolating virus 40(SV40), SV40-LT overexpressed lentivirus vector, was transfected into primary human colorectal adenomatous polyp epithelial cells. The transfected cells were screened, and the screened cells were amplified to obtain the epithelial cell line: IH-CRA-CELL. The cells were identified by morphological observation, cell proliferation, Quantitative real-time PCR (qPCR), and Short Tandem Repeats (STR) experiments. Morphologically, the cells showed epithelial-like characteristics, such as polygon shape, desmosomes mitochondria, and strong positive keratin staining. There was no significant difference between the transfected cells and the primary cells. Through the STR identification experiment, no matching cell lines were found in the cell lines retrieval.

Conclusion

We successfully established a natural LHPP low-expressing precancerous epithelial cell line by SV40-LT antigen gene transfection, which has been patented and is now preserved in the Chinese Typical Culture Preservation Center. It was verified that the transformed cells maintained the phenotype and biological characteristics of epithelial cells. This cell line can be used to study the mechanism of precancerous lesions, screen the efficacy of novel drugs, and construct disease models.

© 2025 Bentham Science Publishers
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2025-04-28
2025-12-23

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/content/journals/pra/10.2174/0115748928297500240522080820
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Construction and Identification of a Human Colorectal Adenoma Epithelial Cell Line by SV40 Large T-antigen Transfection
Recent Patents on Anti-Cancer Drug Discovery 20, 621 (2025); https://doi.org/10.2174/0115748928297500240522080820
/content/journals/pra/10.2174/0115748928297500240522080820
/content/journals/pra/10.2174/0115748928297500240522080820
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  • Article Type:     Research Article
Keyword(s): Colorectal adenoma; epithelial cell line; immortalized; malignant transformation; quantitative real-time PCR; SV40-LT

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