Expression of a Cx43 deletion mutant in 3T3 A31 fibroblasts prevents PDGF-induced inhibition of cell communication and suppresses cell growth

被引:28
作者
Moorby, CD
Gherardi, E
机构
[1] Univ Leicester, Ctr Mechanisms Human Toxic, Leicester LE1 9HN, Leics, England
[2] Univ Cambridge, Ctr Mrc, Dept Oncol, Growth Factors Grp, Cambridge CB2 2QH, England
关键词
gap junctions; cell communication; PDGF; growth regulation;
D O I
10.1006/excr.1999.4485
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
Communication through gap junctions was first suggested to have a role in the social control of cell growth over 30 years ago. However, despite extensive experimentation, the importance of gap junctions as a general mechanism of growth control remains to be established. A number of different studies have shown that a common early response of cells in culture to polypeptide growth factors such as PDGF is a rapid and transient inhibition of cell communication suggesting that a cell may have to lose communication with its neighbors before it can undergo cell division. Here we show that 3T3 A31 fibroblasts exposed to PDGF exhibit a 50% decrease in cell communication as measured by dye transfer in the absence of significant changes in the cellular content and distribution of Cx43. Likewise, PDGF inhibited cell communication in cells transfected either with a vector which did not contain a cDNA or with an expression vector encoding full-length Cx43 fused to a c-myc tag (Cx43-M). In contrast, 3T3 A31 fibroblasts transfected with an expression construct encoding a deletion mutant of Cx43 (Cx43-256M) consisting of amino acids 1-256 of Cx43 fused to a c-myc tag maintain high levels of gap junction activity following exposure to PDG;F. These results suggest that sites which trigger loss of cell communication in response to PDGF are located within amino acids 257 to 382 of the Cx43 molecule. Cells transfected with an expression vector encoding full-length Cx43 fused to a c-myc tail exhibited a reduced basal growth rate compared to both parent cells and cells transfected with a control vector but maintained a strong mitogenic response to PDGF. In contrast, both the basal growth rate and the mitogenic response to PDGF was markedly reduced in cells which expressed Cx43-256M consistent with the hypothesis that loss of cell communication is required before a cell can respond to mitogenic stimuli. (C) 1999 Academic Press.
引用
收藏
页码:367 / 376
页数:10
相关论文
共 32 条
[11]   A particle-receptor model for the insulin-induced closure of connexin43 channels [J].
Homma, N ;
Alvarado, JL ;
Coombs, W ;
Stergiopoulos, K ;
Taffet, SM ;
Lau, AF ;
Delmar, M .
CIRCULATION RESEARCH, 1998, 83 (01) :27-32
[12]   EPIDERMAL GROWTH-FACTOR DISRUPTS GAP-JUNCTIONAL COMMUNICATION AND INDUCES PHOSPHORYLATION OF CONNEXIN43 ON SERINE [J].
LAU, AF ;
KANEMITSU, MY ;
KURATA, WE ;
DANESH, S ;
BOYNTON, AL .
MOLECULAR BIOLOGY OF THE CELL, 1992, 3 (08) :865-874
[13]   A STRUCTURAL BASIS FOR THE UNEQUAL SENSITIVITY OF THE MAJOR CARDIAC AND LIVER GAP-JUNCTIONS TO INTRACELLULAR ACIDIFICATION - THE CARBOXYL TAIL LENGTH [J].
LIU, SG ;
TAFFET, S ;
STONER, L ;
DELMAR, M ;
VALLANO, ML ;
JALIFE, J .
BIOPHYSICAL JOURNAL, 1993, 64 (05) :1422-1433
[14]   PERMEABILITY OF MEMBRANE JUNCTIONS [J].
LOEWENST.WR .
ANNALS OF THE NEW YORK ACADEMY OF SCIENCES, 1966, 137 (A2) :441-+
[15]  
Loewenstein W R, 1992, Semin Cell Biol, V3, P59
[16]   ALTERED REGULATION OF INTERCELLULAR COMMUNICATION BY EPIDERMAL GROWTH-FACTOR, TRANSFORMING GROWTH FACTOR-BETA AND PEPTIDE-HORMONES IN NORMAL HUMAN KERATINOCYTES [J].
MADHUKAR, BV ;
OH, SY ;
CHANG, CC ;
WADE, M ;
TROSKO, JE .
CARCINOGENESIS, 1989, 10 (01) :13-20
[17]   GROWTH-FACTORS MODULATE JUNCTIONAL CELL-TO-CELL COMMUNICATION [J].
MALDONADO, PE ;
ROSE, B ;
LOEWENSTEIN, WR .
JOURNAL OF MEMBRANE BIOLOGY, 1988, 106 (03) :203-210
[18]   GROWTH-INHIBITION OF TRANSFORMED-CELLS CORRELATES WITH THEIR JUNCTIONAL COMMUNICATION WITH NORMAL-CELLS [J].
MEHTA, PP ;
BERTRAM, JS ;
LOEWENSTEIN, WR .
CELL, 1986, 44 (01) :187-196
[19]   INCORPORATION OF THE GENE FOR A CELL-CELL CHANNEL PROTEIN INTO TRANSFORMED-CELLS LEADS TO NORMALIZATION OF GROWTH [J].
MEHTA, PP ;
HOTZWAGENBLATT, A ;
ROSE, B ;
SHALLOWAY, D ;
LOEWENSTEIN, WR .
JOURNAL OF MEMBRANE BIOLOGY, 1991, 124 (03) :207-225
[20]   HGF/SF INHIBITS JUNCTIONAL COMMUNICATION [J].
MOORBY, CD ;
STOKER, M ;
GHERARDI, E .
EXPERIMENTAL CELL RESEARCH, 1995, 219 (02) :657-663