MULTILAYERING AND LOSS OF APICAL POLARITY IN MDCK CELLS TRANSFORMED WITH VIRAL K-RAS

被引:89
作者
SCHOENENBERGER, CA
ZUK, A
KENDALL, D
MATLIN, KS
机构
关键词
D O I
10.1083/jcb.112.5.873
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
The effects of viral Kirsten ras oncogene expression on the polarized phenotype of MDCK cells were investigated. Stable transformed MDCK cell lines expressing the v-K-ras oncogene were generated via infection with a helper-independent retroviral vector construct. When grown on plastic substrata, transformed cells formed continuous monolayers with epithelial-like morphology. However, on permeable filter supports where normal cells form highly polarized monolayers, transformed MDCK cells detached from the substratum and developed multilayers. Morphological analysis of the multilayers revealed that oncogene expression perturbed the polarized organization of MDCK cells such that the transformed cells lacked an apical-basal axis around which the cytoplasm is normally organized. Evidence for selective disruption of apical membrane polarity was provided by immunolocalization of membrane proteins; a normally apical 114-kD protein was randomly distributed on the cell surface in the transformed cell line, whereas normally basolateral proteins remained exclusively localized to areas of cell contact and did not appear on the free cell surface. The discrete distribution of the tight junction-associated ZO-1 protein as well as transepithelial resistance and flux measurements suggested that tight junctions were also assembled. These findings indicate that v-K-ras transformation alters cell-substratum and cell-cell interactions in MDCK cells. Furthermore, v-K-ras expression perturbs apical polarization but does not interfere with the development of a basolateral domain, suggesting that apical and basolateral polarity in epithelial cells may be regulated independently.
引用
收藏
页码:873 / 889
页数:17
相关论文
共 77 条
[21]   JUNCTIONAL COMPLEXES IN VARIOUS EPITHELIA [J].
FARQUHAR, MG ;
PALADE, GE .
JOURNAL OF CELL BIOLOGY, 1963, 17 (02) :375-&
[22]  
FAWCETT DW, 1986, TXB HISTOLOGY, P57
[23]   FROM EGG TO EPITHELIUM [J].
FLEMING, TP ;
JOHNSON, MH .
ANNUAL REVIEW OF CELL BIOLOGY, 1988, 4 :459-485
[24]   TRANSFERRIN RECEPTOR POLARITY AND RECYCLING ACCURACY IN TIGHT AND LEAKY STRAINS OF MADIN-DARBY CANINE KIDNEY-CELLS [J].
FULLER, SD ;
SIMONS, K .
JOURNAL OF CELL BIOLOGY, 1986, 103 (05) :1767-1779
[25]   MONOCLONAL-ANTIBODIES TO THE P21 PRODUCTS OF THE TRANSFORMING GENE OF HARVEY MURINE SARCOMA-VIRUS AND OF THE CELLULAR RAS GENE FAMILY [J].
FURTH, ME ;
DAVIS, LJ ;
FLEURDELYS, B ;
SCOLNICK, EM .
JOURNAL OF VIROLOGY, 1982, 43 (01) :294-304
[26]   ESTABLISHMENT OF 2 RABBIT MAMMARY EPITHELIAL-CELL LINES WITH DISTINCT ONCOGENIC POTENTIAL AND DIFFERENTIATED PHENOTYPE AFTER MICROINJECTION OF TRANSFORMING GENES [J].
GARCIA, I ;
SORDAT, B ;
RAUCCIOFARINON, E ;
DUNAND, M ;
KRAEHENBUHL, JP ;
DIGGELMANN, H .
MOLECULAR AND CELLULAR BIOLOGY, 1986, 6 (06) :1974-1982
[27]   NEW TECHNIQUE FOR ASSAY OF INFECTIVITY OF HUMAN ADENOVIRUS 5 DNA [J].
GRAHAM, FL ;
VANDEREB, AJ .
VIROLOGY, 1973, 52 (02) :456-467
[28]   STRUCTURE, BIOCHEMISTRY, AND ASSEMBLY OF EPITHELIAL TIGHT JUNCTIONS [J].
GUMBINER, B .
AMERICAN JOURNAL OF PHYSIOLOGY, 1987, 253 (06) :C749-C758
[29]   THE ROLE OF THE CELL-ADHESION MOLECULE UVOMORULIN IN THE FORMATION AND MAINTENANCE OF THE EPITHELIAL JUNCTIONAL COMPLEX [J].
GUMBINER, B ;
STEVENSON, B ;
GRIMALDI, A .
JOURNAL OF CELL BIOLOGY, 1988, 107 (04) :1575-1587
[30]   A FUNCTIONAL ASSAY FOR PROTEINS INVOLVED IN ESTABLISHING AN EPITHELIAL OCCLUDING BARRIER - IDENTIFICATION OF A UVOMORULIN-LIKE POLYPEPTIDE [J].
GUMBINER, B ;
SIMONS, K .
JOURNAL OF CELL BIOLOGY, 1986, 102 (02) :457-468