TP53 Mutation by CRISPR System Enhances the Malignant Potential of Colon Cancer.

Abstract

Tumor protein p53 (TP53) mutation is a well-known occurrence at the late phase of carcinogenesis during the adenoma-carcinoma sequence of a sporadic colon cancer. Although numerous reports about clinical information of the patients with colon cancer have suggested that TP53 mutation might be related to various types of malignant potential, the direct effects of this mutation on the malignant potential of colon cancer remain unknown. Notably, no previous report has described a relationship between TP53 mutation and cancer stemness. We therefore aimed to assess the function of a TP53 mutant induced by the CRISPR-Cas9 system in colon cancer cells. In this study, two TP53 mutations, corresponding to exon 3 (TP53E3) and 10 (TP53E10), were generated in LS174T cells derived from a wild-type TP53 human colon cancer via a lentiviral CRISPR-Cas9 system. The loss of function of TP53 resulting from both mutations manifested as resistance to Nutlin3a-induced apoptosis and the downregulation of target genes of TP53. TP53 mutants exhibited an enhanced malignant potential, characterized by accelerated cell growth, invasiveness, chemoresistance, and cancer stemness. Interestingly, TP53E10 but not TP53E3 cells exhibited aberrant transcriptional activity of regenerating family member 1-α (REG1A) and expression of REG1A, resulting in the acquisition of enhanced malignant potential. In conclusion, we demonstrated for the first time that TP53 genomic mutation into human colon cancer cells affects the malignant potential. IMPLICATIONS: These findings suggest that both a loss of function and an aberrant gain of function of TP53 might promote high malignant potentials at the late phase of carcinogenesis in colon cancer.

• Authors: Sho Watanabe, Kiichiro Tsuchiya, Ryu Nishimura, Tomoaki Shirasaki, Nobuhiro Katsukura, Shuji Hibiya, Ryuichi Okamoto, Tetsuya Nakamura, Mamoru Watanabe
• Journal: Molecular cancer research : MCR (Mol Cancer Res) Vol. 17 Issue 7 Pg. 1459-1467 (07 2019) ISSN: 1557-3125 [Electronic] United States
• PMID: 30988165 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: ©2019 American Association for Cancer Research.

Chemical References

• Lithostathine
• REG1A protein, human
• TP53 protein, human
• Tumor Suppressor Protein p53

Topics

• CRISPR-Cas Systems (genetics)
• Carcinogenesis (genetics)
• Cell Line, Tumor
• Cell Proliferation (genetics)
• Colonic Neoplasms (drug therapy, genetics, pathology)
• Drug Resistance, Neoplasm (genetics)
• Exons (genetics)
• Gene Expression Regulation, Neoplastic
• Humans
• Lithostathine (genetics)
• Mutation (genetics)
• Neoplasm Invasiveness (genetics, pathology)
• Neoplastic Stem Cells (metabolism, pathology)
• Tumor Suppressor Protein p53 (genetics)