title: Cell type-specific targeted mutations of Kras and Pten document proliferation arrest in granulosa cells versus oncogenic insult to ovarian surface epithelial cells.
abstract: The small G-protein KRAS is crucial for mediating gonadotropin-induced events associated with ovulation. However, constitutive expression of KrasG12D in granulosa cells disrupted normal follicle development leading to the persistence of abnormal follicle-like structures containing nonmitotic cells. To determine what factors mediate this potent effect of KrasG12D, gene profiling analyses were done. We also analyzed KrasG12D;Cyp19-Cre and KrasG12;Pgr-Cre mutant mouse models that express Cre prior to or after the initiation of granulosa cell differentiation, respectively. KrasG12D induced cell cycle arrest in granulosa cells of the KrasG12D;Cyp19-Cre mice but not in the KrasG12D;Pgr-Cre mice, documenting the cell context-specific effect of KrasG12D. Expression of KrasG12D silenced the Kras gene, reduced cell cycle activator genes, and impaired the expression of granulosa cell and oocyte-specific genes. Conversely, levels of PTEN and phosphorylated p38 mitogen-activated protein kinase (MAPK) increased markedly in the mutant granulosa cells. Because disrupting Pten in granulosa cells leads to increased proliferation and survival, Pten was disrupted in the KrasG12D mutant mice. The Pten/Kras mutant mice were infertile but lacked granulosa cell tumors. By contrast, the Ptenfl/fl;KrasG12D;Amhr2-Cre mice developed aggressive ovarian surface epithelial cell tumors that did not occur in the Ptenfl/fl;KrasG12D;Cyp19-Cre or Ptenfl/fl;KrasG12D;Pgr-Cre mouse strains. These data document unequivocally that Amhr2-Cre is expressed in and mediates allelic recombination of oncogenic genes in ovarian surface epithelial cells. That KrasG12D/Pten mutant granulosa cells do not transform but rather undergo cell cycle arrest indicates that they resist the oncogenic insults of Kras/Pten by robust self-protecting mechanisms that silence the Kras gene and elevate PTEN and phosphorylated p38 MAPK.
authors.affiliation: Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas 77030, USA.
authors.forename: Heng-Yu
authors.initials: HY
authors.lastname: Fan
authors.forename: Zhilin
authors.initials: Z
authors.lastname: Liu
authors.forename: Marilene
authors.initials: M
authors.lastname: Paquet
authors.forename: Jinrong
authors.initials: J
authors.lastname: Wang
authors.forename: John P
authors.initials: JP
authors.lastname: Lydon
authors.forename: Francesco J
authors.initials: FJ
authors.lastname: DeMayo
authors.forename: JoAnne S
authors.initials: JS
authors.lastname: Richards
mesh_terms: D000818:Animals; D002453:Cell Cycle / Q000235:genetics; D049109:Cell Proliferation; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D004847:Epithelial Cells / Q000378:metabolism / Q000473:pathology*; D005260:Female; D011905:Genes, ras; D006107:Granulosa Cells / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D008954:Models, Biological; D016297:Mutagenesis, Site-Directed; D009154:Mutation / Q000502:physiology; D009928:Organ Specificity / Q000235:genetics; D010053:Ovary / Q000378:metabolism / Q000473:pathology; D051059:PTEN Phosphohydrolase / Q000235:genetics* / Q000378:metabolism; D010766:Phosphorylation; D016283:Proto-Oncogene Proteins p21(ras) / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
chemical_list: D048051:p38 Mitogen-Activated Protein Kinases; D051059:PTEN Phosphohydrolase; C487757:Pten protein, mouse; C495971:Hras protein, mouse; D016283:Proto-Oncogene Proteins p21(ras)
references.citation: Proc Natl Acad Sci U S A. 2006 Jul 25;103(30):11154-9
references.pmid: 16849428
references.citation: Nat Genet. 2004 Jun;36(6):653-9
references.pmid: 15156143
references.citation: Mol Endocrinol. 2008 Sep;22(9):2128-40
references.pmid: 18606860
references.citation: Cancer Cell. 2004 Apr;5(4):375-87
references.pmid: 15093544
references.citation: Nat Genet. 2007 Jun;39(6):741-9
references.pmid: 17468757
references.citation: Curr Opin Cell Biol. 2001 Dec;13(6):748-53
references.pmid: 11698192
references.citation: Differentiation. 2008 Dec;76(10):1081-92
references.pmid: 18637025
references.citation: Mol Endocrinol. 2007 Aug;21(8):1940-57
references.pmid: 17536007
references.citation: Mol Cell Biol. 2003 Dec;23(24):9375-88
references.pmid: 14645547
references.citation: Cancer Cell. 2007 Apr;11(4):321-33
references.pmid: 17418409
references.citation: Nat Med. 2005 Jan;11(1):63-70
references.pmid: 15619626
references.citation: Cancer Treat Res. 2002;107:293-302
references.pmid: 11775457
references.citation: Cancer Cell. 2006 Dec;10(6):459-72
references.pmid: 17157787
references.citation: Mol Cell Biol. 2008 Mar;28(5):1770-82
references.pmid: 18172011
references.citation: Trends Endocrinol Metab. 2000 Oct;11(8):309-14
references.pmid: 10996525
references.citation: J Biol Chem. 2004 May 7;279(19):19431-40
references.pmid: 14982927
references.citation: Genesis. 2005 Feb;41(2):58-66
references.pmid: 15682389
references.citation: Carcinogenesis. 2008 Nov;29(11):2062-72
references.pmid: 18687666
references.citation: J Clin Oncol. 2004 Jul 15;22(14):2954-63
references.pmid: 15254063
references.citation: Clin Cancer Res. 2006 Mar 1;12(5):1593-8
references.pmid: 16533786
references.citation: Nature. 2007 Oct 25;449(7165):1073-7
references.pmid: 17960246
references.citation: Recent Prog Horm Res. 1995;50:223-54
references.pmid: 7740159
references.citation: Cancer Res. 2003 Mar 15;63(6):1389-97
references.pmid: 12649204
references.citation: Cell. 1997 Mar 7;88(5):593-602
references.pmid: 9054499
references.citation: Biol Reprod. 1995 Jul;53(1):103-9
references.pmid: 7669840
references.citation: Nature. 2001 Apr 26;410(6832):1111-6
references.pmid: 11323676
references.citation: Cancer Res. 2006 Sep 15;66(18):8949-53
references.pmid: 16982732
references.citation: Cell. 2007 Jun 1;129(5):957-68
references.pmid: 17540175
references.citation: PLoS Biol. 2008 Dec 2;6(12):2853-68
references.pmid: 19053174
references.citation: Nat Genet. 2004 Jun;36(6):647-52
references.pmid: 15156142
references.citation: Nat Genet. 1998 Aug;19(4):348-55
references.pmid: 9697695
references.citation: Genes Cells. 2003 Feb;8(2):131-44
references.pmid: 12581156
references.citation: Nat Genet. 2002 Nov;32(3):408-10
references.pmid: 12368913
references.citation: Mol Cell. 2007 Jul 6;27(1):107-19
references.pmid: 17612494
references.citation: Trends Cell Biol. 2003 Sep;13(9):478-83
references.pmid: 12946627
references.citation: Mol Endocrinol. 2006 Jun;20(6):1300-21
references.pmid: 16455817
references.citation: Nature. 1992 Nov 26;360(6402):313-9
references.pmid: 1448148
references.citation: Science. 2009 May 15;324(5929):938-41
references.pmid: 19443782
references.citation: Mol Endocrinol. 2006 Dec;20(12):3308-20
references.pmid: 16973759
references.citation: Mol Endocrinol. 2000 Aug;14(8):1283-300
references.pmid: 10935551
references.citation: Mol Cell Endocrinol. 2000 May 25;163(1-2):61-6
references.pmid: 10963875
references.citation: Reprod Biol Endocrinol. 2003 Oct 07;1:67
references.pmid: 14613552
references.citation: Science. 2005 Aug 5;309(5736):886-7
references.pmid: 16081723
references.citation: Nat Cell Biol. 2007 May;9(5):493-505
references.pmid: 17450133
references.citation: Mol Cell. 2007 Jul 20;27(2):176-7
references.pmid: 17643366
references.citation: Genesis. 2002 Feb;32(2):148-9
references.pmid: 11857804
references.citation: Development. 2008 Jun;135(12):2127-37
references.pmid: 18506027
references.citation: Proc Natl Acad Sci U S A. 2008 Aug 26;105(34):12469-73
references.pmid: 18711140
references.citation: Biochim Biophys Acta. 2000 Feb 14;1470(1):M21-35
references.pmid: 10656987
references.citation: Int J Gynecol Pathol. 2008 Apr;27(2):151-60
references.pmid: 18317228