title: Mutant p53 cooperates with knockdown of endogenous wild-type p53 to disrupt tubulogenesis in Madin-Darby canine kidney cells.
abstract: Mutation of the p53 gene is the most common genetic alteration in human malignances and associated clinically with tumor progression and metastasis. To determine the effect of mutant p53 on epithelial differentiation, we developed three-dimensional culture (3-D) of Madin-Darby canine kidney (MDCK) cells. We found that parental MDCK cells undergo a series of morphological changes and form polarized and growth-arrested cysts with hollow lumen, which resembles branching tubules in vitro. We also found that upon knockdown of endogenous wild-type p53 (p53-KD), MDCK cells still form normal cysts in 3-D culture, indicating that p53-KD alone is not sufficient to disrupt cysts formation. However, we found that ectopic expression of mutant R163H (human equivalent R175H) or R261H (human equivalent R273H) in MDCK cells leads to disruption of cyst polarity and formation of invasive aggregates, which is further compounded by knockdown of endogenous wild-type p53. Consistently, we found that expression of E-cadherin, β-catenin, and epithelial-to-mesenchymal transition (EMT) transcription factors (Snail-1, Slug and Twist) is altered by mutant p53, which is also compounded by knockdown of wild-type p53. Moreover, the expression level of c-Met, the hepatocyte growth factor receptor and a key regulator of kidney cell tubulogenesis, is enhanced by combined knockdown of endogenous wild-type p53 and ectopic expression of mutant R163H or R261H but not by each individually. Together, our data suggest that upon inactivating mutation of the p53 gene, mutant p53 acquires its gain of function by altering morphogenesis and promoting cell migration and invasion in part by upregulating EMT and c-Met.
authors.affiliation: Comparative Oncology Laboratory, Schools of Medicine and Veterinary Medicine, University of California Davis, Davis, California, United States of America.
authors.forename: Yanhong
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Comparative Oncology Laboratory, Schools of Medicine and Veterinary Medicine, University of California Davis, Davis, California, United States of America.
authors.forename: Wensheng
authors.initials: W
authors.lastname: Yan
authors.affiliation: Comparative Oncology Laboratory, Schools of Medicine and Veterinary Medicine, University of California Davis, Davis, California, United States of America.
authors.forename: Xinbin
authors.initials: X
authors.lastname: Chen
mesh_terms: D000818:Animals; D002465:Cell Movement; D004285:Dogs; D058750:Epithelial-Mesenchymal Transition; D055785:Gene Knockdown Techniques; D007684:Kidney Tubules / Q000378:metabolism*; D061985:Madin Darby Canine Kidney Cells; D020125:Mutation, Missense; D016159:Tumor Suppressor Protein p53 / Q000276:immunology / Q000378:metabolism*
chemical_list: D016159:Tumor Suppressor Protein p53