CINP is a novel cofactor of KLF5 required for its role in the promotion of cell proliferation, survival and tumor growth

Krüppel‐like factor 5 (KLF5) both suppresses and promotes tumor growth depending on cellular context. The mechanisms underlying tumor promotion could be targetable for therapy. Although a number of transcriptional targets of KLF5 have been identified and implicated in KLF5‐mediated tumor growth, how KLF5 regulates these genes remains to be addressed. Here we performed coimmunoprecipitation (co‐IP) and liquid chromatography–tandem mass spectrometry (LC–MS/MS) in the TSU‐Pr1 bladder cancer cell line, in which KLF5 is shown to promote tumor growth, to identify KLF5‐interacting nuclear proteins that are necessary for KLF5’s tumor promoting function. LC–MS/MS revealed 122 potential KLF5 binding proteins in the nuclear proteins precipitated by the KLF5 antibody, and the top nine candidates included AHNAK, TFAM, HSDL2, HNRNPC, CINP, IST1, FBL, PABPC1 and SNRNP40. SRB assays of these nine proteins indicated that silencing CINP had the most potent inhibitory effect on cell growth in KLF5‐expressing cells but did not affect parental TSU‐Pr1 cells. Further analyses not only confirmed the physical interaction between KLF5 and CINP, also demonstrated that knockdown of CINP attenuated the effects of KLF5 on cell cycle progression, apoptosis and tumorigenesis. Silencing CINP also attenuated the effect of KLF5 on the expression of a number of genes and signaling pathways, including cell cycle regulator Cyclin D1 and apoptosis‐related Caspase 7. These results suggest that CINP is a cofactor of KLF5 that is crucial for the promotion of tumor growth, and that the KLF5‐CINP interaction could be a novel therapeutic target for inhibiting KLF5‐promoted tumor growth. What's new? The role of Krüppel‐like factor 5 (KLF5) in promoting cell proliferation and tumorigenesis is well‐established, but although several KLF5 transcriptional targets have been identified, how KLF5 regulates these genes remains elusive. Here, using immunoprecipitation combined with mass spectrometry, the authors identified a number of KLF5‐interacting nuclear proteins. Functional screening revealed that the Cdk2‐interacting protein CINP is required for KLF5’s tumor promoting function and a KLF5 cofactor. CINP knockdown attenuated the effects of KLF5 on cell and tumor growth by decelerating cell cycle progression and inducing apoptosis in KLF5‐expressing cells. The KLF5‐CINP interaction provides a therapeutic opportunity for targeting KLF5‐mediated tumorigenesis.