Growth-suppressive function of human connexin32 in a conditional immortalized mouse hepatocyte cell line

T. Kojima, M. Srinivas, A. Fort, M. Urban, G. H. Lee, N. Sawada, David C. Spray

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Mouse hepatocytes immortalized with a temperature-sensitive allele of the SV40 large T-antigen (CHST8 cells) were found to lack the high expression of the gap junction proteins Cx26 and Cx32 that characterizes normal mouse hepatocytes, expressing instead Cx43 and Cx45 at minimal levels. In order to examine the growth suppressive function of Cx32 on hepatocytes, we transfected these CHST8 cells with human Cx32 complementary deoxyribonucleic acid and measured the growth rates at 33, 37, and 39° C. Expression of human Cx32 and its messenger ribonucleic acid in the stable cell lines was confirmed by immunocytochemistry and by Western and Northern blots analyses. Dye transfer following lucifer yellow injection into the transfectants was extensive; Cx32 channels displayed unitary conductances of about 70 pS and were moderately voltage sensitive. When cultured at 33 and 39° C, growth rates of both parental cells and transfectants were of the same level. When examined at 37° C, growth rate of the transfectant, which highly expressed Cx32 at the membranes, was significantly decreased compared to the parental cells. However, no changes in the expression of Cx32 protein in the transfectants were observed between 33 and 37° C. These results suggest that Cx32 expression could inhibit hepatocyte growth in vitro using the conditional immortalized cells. Cx32 transfectants using a conditional immortalized mouse hepatocyte may be useful for examining the mechanisms of growth and differentiation in hepatocytes by gap junction expression.

Original languageEnglish (US)
Pages (from-to)589-598
Number of pages10
JournalIn Vitro Cellular and Developmental Biology - Animal
Volume37
Issue number9
DOIs
StatePublished - 2001

Fingerprint

Hepatocytes
Cells
Cell Line
Growth
Polyomavirus Transforming Antigens
Connexin 43
Connexins
Gap Junctions
Viral Tumor Antigens
Northern Blotting
Coloring Agents
Western Blotting
Immunohistochemistry
Alleles
RNA
Membranes
Injections
Temperature
DNA
Electric potential

Keywords

  • Cell growth
  • Cx32
  • Mouse hepatocytes

ASJC Scopus subject areas

  • Developmental Biology
  • Clinical Biochemistry
  • Cell Biology

Cite this

Growth-suppressive function of human connexin32 in a conditional immortalized mouse hepatocyte cell line. / Kojima, T.; Srinivas, M.; Fort, A.; Urban, M.; Lee, G. H.; Sawada, N.; Spray, David C.

In: In Vitro Cellular and Developmental Biology - Animal, Vol. 37, No. 9, 2001, p. 589-598.

Research output: Contribution to journalArticle

Kojima, T. ; Srinivas, M. ; Fort, A. ; Urban, M. ; Lee, G. H. ; Sawada, N. ; Spray, David C. / Growth-suppressive function of human connexin32 in a conditional immortalized mouse hepatocyte cell line. In: In Vitro Cellular and Developmental Biology - Animal. 2001 ; Vol. 37, No. 9. pp. 589-598.
@article{8c1b20dd365740d5b12e07a31ef544e7,
title = "Growth-suppressive function of human connexin32 in a conditional immortalized mouse hepatocyte cell line",
abstract = "Mouse hepatocytes immortalized with a temperature-sensitive allele of the SV40 large T-antigen (CHST8 cells) were found to lack the high expression of the gap junction proteins Cx26 and Cx32 that characterizes normal mouse hepatocytes, expressing instead Cx43 and Cx45 at minimal levels. In order to examine the growth suppressive function of Cx32 on hepatocytes, we transfected these CHST8 cells with human Cx32 complementary deoxyribonucleic acid and measured the growth rates at 33, 37, and 39° C. Expression of human Cx32 and its messenger ribonucleic acid in the stable cell lines was confirmed by immunocytochemistry and by Western and Northern blots analyses. Dye transfer following lucifer yellow injection into the transfectants was extensive; Cx32 channels displayed unitary conductances of about 70 pS and were moderately voltage sensitive. When cultured at 33 and 39° C, growth rates of both parental cells and transfectants were of the same level. When examined at 37° C, growth rate of the transfectant, which highly expressed Cx32 at the membranes, was significantly decreased compared to the parental cells. However, no changes in the expression of Cx32 protein in the transfectants were observed between 33 and 37° C. These results suggest that Cx32 expression could inhibit hepatocyte growth in vitro using the conditional immortalized cells. Cx32 transfectants using a conditional immortalized mouse hepatocyte may be useful for examining the mechanisms of growth and differentiation in hepatocytes by gap junction expression.",
keywords = "Cell growth, Cx32, Mouse hepatocytes",
author = "T. Kojima and M. Srinivas and A. Fort and M. Urban and Lee, {G. H.} and N. Sawada and Spray, {David C.}",
year = "2001",
doi = "10.1290/1071-2690(2001)037<0589:GSFOHC>2.0.CO;2",
language = "English (US)",
volume = "37",
pages = "589--598",
journal = "In Vitro Cellular and Developmental Biology - Plant",
issn = "1054-5476",
publisher = "Springer New York",
number = "9",

}

TY - JOUR

T1 - Growth-suppressive function of human connexin32 in a conditional immortalized mouse hepatocyte cell line

AU - Kojima, T.

AU - Srinivas, M.

AU - Fort, A.

AU - Urban, M.

AU - Lee, G. H.

AU - Sawada, N.

AU - Spray, David C.

PY - 2001

Y1 - 2001

N2 - Mouse hepatocytes immortalized with a temperature-sensitive allele of the SV40 large T-antigen (CHST8 cells) were found to lack the high expression of the gap junction proteins Cx26 and Cx32 that characterizes normal mouse hepatocytes, expressing instead Cx43 and Cx45 at minimal levels. In order to examine the growth suppressive function of Cx32 on hepatocytes, we transfected these CHST8 cells with human Cx32 complementary deoxyribonucleic acid and measured the growth rates at 33, 37, and 39° C. Expression of human Cx32 and its messenger ribonucleic acid in the stable cell lines was confirmed by immunocytochemistry and by Western and Northern blots analyses. Dye transfer following lucifer yellow injection into the transfectants was extensive; Cx32 channels displayed unitary conductances of about 70 pS and were moderately voltage sensitive. When cultured at 33 and 39° C, growth rates of both parental cells and transfectants were of the same level. When examined at 37° C, growth rate of the transfectant, which highly expressed Cx32 at the membranes, was significantly decreased compared to the parental cells. However, no changes in the expression of Cx32 protein in the transfectants were observed between 33 and 37° C. These results suggest that Cx32 expression could inhibit hepatocyte growth in vitro using the conditional immortalized cells. Cx32 transfectants using a conditional immortalized mouse hepatocyte may be useful for examining the mechanisms of growth and differentiation in hepatocytes by gap junction expression.

AB - Mouse hepatocytes immortalized with a temperature-sensitive allele of the SV40 large T-antigen (CHST8 cells) were found to lack the high expression of the gap junction proteins Cx26 and Cx32 that characterizes normal mouse hepatocytes, expressing instead Cx43 and Cx45 at minimal levels. In order to examine the growth suppressive function of Cx32 on hepatocytes, we transfected these CHST8 cells with human Cx32 complementary deoxyribonucleic acid and measured the growth rates at 33, 37, and 39° C. Expression of human Cx32 and its messenger ribonucleic acid in the stable cell lines was confirmed by immunocytochemistry and by Western and Northern blots analyses. Dye transfer following lucifer yellow injection into the transfectants was extensive; Cx32 channels displayed unitary conductances of about 70 pS and were moderately voltage sensitive. When cultured at 33 and 39° C, growth rates of both parental cells and transfectants were of the same level. When examined at 37° C, growth rate of the transfectant, which highly expressed Cx32 at the membranes, was significantly decreased compared to the parental cells. However, no changes in the expression of Cx32 protein in the transfectants were observed between 33 and 37° C. These results suggest that Cx32 expression could inhibit hepatocyte growth in vitro using the conditional immortalized cells. Cx32 transfectants using a conditional immortalized mouse hepatocyte may be useful for examining the mechanisms of growth and differentiation in hepatocytes by gap junction expression.

KW - Cell growth

KW - Cx32

KW - Mouse hepatocytes

UR - http://www.scopus.com/inward/record.url?scp=0034754975&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034754975&partnerID=8YFLogxK

U2 - 10.1290/1071-2690(2001)037<0589:GSFOHC>2.0.CO;2

DO - 10.1290/1071-2690(2001)037<0589:GSFOHC>2.0.CO;2

M3 - Article

VL - 37

SP - 589

EP - 598

JO - In Vitro Cellular and Developmental Biology - Plant

JF - In Vitro Cellular and Developmental Biology - Plant

SN - 1054-5476

IS - 9

ER -