Inhibition of gap junctional intercellular communication and enhancement of growth in BALB/c 3t3 cells treated with connexin43 antisense oligonucleotides

✍ Scribed by Randall J. Ruch; Xiaojun Guan; Kristi Sigler

• Publisher: John Wiley and Sons
• Year: 1995
• Tongue: English
• Weight: 671 KB
• Volume: 14
• Category: Article
• ISSN: 0899-1987
• DOI: 10.1002/mc.2940140407

✦ Synopsis

Abstract

Many studies have correlated reductions in gap junctional intercellular communication (a) with altered cellular growth, tumor promotion, and neoplastic transformation. To test directly whether reduced GJIC affects cellular growth, GJIC was inhibited in murine BALB/c 3T3 fibroblasts by treatment with a phosphorothioatemodified antisense oligonucleotide targeted against the connexin43 translation start codon, and in vitro cell growth was monitored. The cells were incubated with the oligonucleotide (0.1‐0.5 μM) in liposomes in serumless culture medium for 16 h; washed and refed with serum‐containing medium; and analyzed for dye‐coupling, connexin43 protein and mRNA levels, and cell growth over the next 5 d. The antisense oligonucleotide inhibited dye‐coupling and reduced connexin43 protein levels in a concentration‐dependent manner but had no effect on connexin43 mRNA levels. Cell growth rate was not affected, but saturation density was increased approximately threefold by the oligonucleotide. These data support a role for GJIC in the establishment of contact inhibition of in vitro cell growth. © 1995 Wiley‐ Liss, Inc.