00085472can141494-sup-131321_1_supp_2837116_n9ssnh.ppt (219.5 kB)

Supplemental Figure 2 from CDK2 Inhibition Causes Anaphase Catastrophe in Lung Cancer through the Centrosomal Protein CP110

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posted on 2023-03-30, 23:06 authored by Shanhu Hu, Alexey V. Danilov, Kristina Godek, Bernardo Orr, Laura J. Tafe, Jaime Rodriguez-Canales, Carmen Behrens, Barbara Mino, Cesar A. Moran, Vincent A. Memoli, Lisa Maria Mustachio, Fabrizio Galimberti, Saranya Ravi, Andrew DeCastro, Yun Lu, David Sekula, Angeline S. Andrew, Ignacio I. Wistuba, Sarah Freemantle, Duane A. Compton, Ethan Dmitrovsky

Supplemental Fig. S2. Overexpression of wild-type CP110, but not phosphorylation-sites mutated CP110 in LKR13 murine lung cancer cells that are driven by KRAS rescued anaphase catastrophe and reduced apoptosis induced by pharmacological CDK2 inhibition after seliciclib treatment. LKR13 cells overexpressing wild-type CP110 species or an empty vector, a phosphorylation-sites mutant-CP110 species (MUT-CP110) or an empty vector (Vector) were treated with indicated dosages of seliciclib. After 24 hours treatment, LKR13 cells were (A) scored for multipolar anaphase and (B) analyzed for apoptosis, as detected by Annexin V:FITC and 7-aminoactinomycin D (7-AAD) staining.

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ARTICLE ABSTRACT

Aneuploidy is frequently detected in human cancers and is implicated in carcinogenesis. Pharmacologic targeting of aneuploidy is an attractive therapeutic strategy, as this would preferentially eliminate malignant over normal cells. We previously discovered that CDK2 inhibition causes lung cancer cells with more than two centrosomes to undergo multipolar cell division leading to apoptosis, defined as anaphase catastrophe. Cells with activating KRAS mutations were especially sensitive to CDK2 inhibition. Mechanisms of CDK2-mediated anaphase catastrophe and how activated KRAS enhances this effect were investigated. Live-cell imaging provided direct evidence that following CDK2 inhibition, lung cancer cells develop multipolar anaphase and undergo multipolar cell division with the resulting progeny apoptotic. The siRNA-mediated repression of the CDK2 target and centrosome protein CP110 induced anaphase catastrophe of lung cancer cells. In contrast, CP110 overexpression antagonized CDK2 inhibitor–mediated anaphase catastrophe. Furthermore, activated KRAS mutations sensitized lung cancer cells to CDK2 inhibition by deregulating CP110 expression. Thus, CP110 is a critical mediator of CDK2 inhibition–driven anaphase catastrophe. Independent examination of murine and human paired normal–malignant lung tissues revealed marked upregulation of CP110 in malignant versus normal lung. Human lung cancers with KRAS mutations had significantly lower CP110 expression as compared with KRAS wild-type cancers. Thus, a direct link was found between CP110 and CDK2 inhibitor antineoplastic response. CP110 plays a mechanistic role in response of lung cancer cells to CDK2 inhibition, especially in the presence of activated KRAS mutations. Cancer Res; 75(10); 2029–38. ©2015 AACR.