Mirk regulates the exit of colon cancer cells from quiescence.
Mirk/Dyrk1B is a serine/threonine kinase widely expressed in colon cancers. Serum starvation induced HD6 colon carcinoma cells to enter a quiescent G0 state, characterized by a 2N DNA content and a lower RNA content than G1 cells. Compared with cycling cells, quiescent cells exhibited 16-fold higher levels of the retinoblastoma protein p130/Rb2, which sequesters E2F4 to block entry into G1, 10-fold elevated levels of the CDK inhibitor p27kip1, and 10-fold higher levels of Mirk. However, depletion of Mirk did not prevent entry into G0, but enabled quiescent HD6, SW480, and colo320 colon carcinoma cells to acquire some biochemical characteristics of G1 cells, including increased levels of cyclin D1 and cyclin D3 because of slower turnover, increased activity of their CDK4/cyclin D complexes, and increased phosphorylation and decreased E2F4 sequestering ability of the CDK4 target, p130/Rb2. As a result, depletion of Mirk allowed some cells to escape quiescence and enabled cells released from quiescence to traverse G1 more quickly. The kinase activity of Mirk was increased by the chemotherapeutic drug 5-fluorouracil (5-FU). Treatment of p53 mutant colon cancer cells with 5-FU led to an elongated G1 in a Mirk-dependent manner, as G1 was shortened by ectopic overexpression of cyclin D1 mutated at the Mirk phosphorylation site (T288A), but not by wild-type cyclin D1. Mirk, through regulating cyclin D turnover, and the CDK inhibitor p27, as shown by depletion studies, functioned independently and additively to regulate the exit of tumor cells from quiescence.
Jin
Kideok
K
Pathology Department, Upstate Medical University of the State University of New York, Syracuse, New York 13210, USA.
Ewton
Daina Z
DZ
Park
Sunju
S
Hu
Jing
J
Friedman
Eileen
E
D000963:Antimetabolites / Q000494:pharmacology; D002453:Cell Cycle / Q000187:drug effects / Q000235:genetics; D002460:Cell Line; D003110:Colonic Neoplasms / Q000378:metabolism* / Q000473:pathology; D056741:Cyclin D; D050760:Cyclin-Dependent Kinase Inhibitor p27 / Q000235:genetics / Q000502:physiology; D016213:Cyclins / Q000378:metabolism; D004171:Diploidy; D004789:Enzyme Activation / Q000187:drug effects; D005434:Flow Cytometry; D005472:Fluorouracil / Q000494:pharmacology; D019073:HT29 Cells; D006801:Humans; D047468:Immunoprecipitation; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D055550:Protein Stability; D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000502:physiology; D000097603:Dyrk Kinases
D000963:Antimetabolites; D056741:Cyclin D; D050760:Cyclin-Dependent Kinase Inhibitor p27; D016213:Cyclins; D005472:Fluorouracil; D017346:Protein Serine-Threonine Kinases; D011505:Protein-Tyrosine Kinases; D016159:Tumor Suppressor Protein p53; D000097603:Dyrk Kinases