The cytochrome P450 enzyme CYP24A1 increases proliferation of mutant KRAS-dependent lung adenocarcinoma independent of its catalytic activity.

Abstract

We previously reported that overexpression of cytochrome P450 family 24 subfamily A member 1 (CYP24A1) increases lung cancer cell proliferation by activating RAS signaling and that CYP24A1 knockdown inhibits tumor growth. However, the mechanism of CYP24A1-mediated cancer cell proliferation remains unclear. Here, we conducted cell synchronization and biochemical experiments in lung adenocarcinoma cells, revealing a link between CYP24A1 and anaphase-promoting complex (APC), a key cell cycle regulator. We demonstrate that CYP24A1 expression is cell cycle-dependent; it was higher in the G2-M phase and diminished upon G1 entry. CYP24A1 has a functional destruction box (D-box) motif that allows binding with two APC adaptors, CDC20-homologue 1 (CDH1) and cell division cycle 20 (CDC20). Unlike other APC substrates, however, CYP24A1 acted as a pseudo-substrate, inhibiting CDH1 activity and promoting mitotic progression. Conversely, overexpression of a CYP24A1 D-box mutant compromised CDH1 binding, allowing CDH1 hyperactivation, thereby hastening degradation of its substrates cyclin B1 and CDC20, and accumulation of the CDC20 substrate p21, prolonging mitotic exit. These activities also occurred with a CYP24A1 isoform 2 lacking the catalytic cysteine (Cys-462), suggesting that CYP24A1's oncogenic potential is independent of its catalytic activity. CYP24A1 degradation reduced clonogenic survival of mutant KRAS-driven lung cancer cells, and calcitriol treatment increased CYP24A1 levels and tumor burden in Lsl-KRASG12D mice. These results disclose a catalytic activity-independent growth-promoting role of CYP24A1 in mutant KRAS-driven lung cancer. This suggests that CYP24A1 could be therapeutically targeted in lung cancers in which its expression is high.

• Authors: Wei Huang, Paramita Ray, Wenbin Ji, Zhuwen Wang, Derek Nancarrow, Guoan Chen, Stefanie Galbán, Theodore S Lawrence, David G Beer, Alnawaz Rehemtulla, Nithya Ramnath, Dipankar Ray
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 295 Issue 18 Pg. 5906-5917 (05 01 2020) ISSN: 1083-351X [Electronic] United States
• PMID: 32165494 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, U.S. Gov't, Non-P.H.S.)

Chemical References

• KRAS protein, human
• Vitamin D3 24-Hydroxylase
• Proto-Oncogene Proteins p21(ras)

Topics

• Adenocarcinoma of Lung (genetics, pathology)
• Biocatalysis
• Cell Cycle
• Cell Line, Tumor
• Cell Proliferation (genetics)
• Cell Survival
• Gene Expression Regulation, Neoplastic
• Humans
• Mutation
• Proto-Oncogene Proteins p21(ras) (genetics)
• Up-Regulation
• Vitamin D3 24-Hydroxylase (genetics, metabolism)