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  • Title: Increased genetic stability of HeLa cells after connexin 43 gene transfection.
    Author: Zhu W, Mironov N, Yamasaki H.
    Journal: Cancer Res; 1997 Jun 01; 57(11):2148-50. PubMed ID: 9187113.
    Abstract:
    To test the hypothesis that intact gap-junctional intercellular communication (GJIC) is necessary for genomic stability, we compared the spontaneous and chemically induced mutation frequencies in GJIC-proficient and -deficient HeLa cells. Thus, we determined microsatellite instability and mutation frequency in the HPRT gene in parental HeLa cells, which have no GJIC ability, and in HeLa cells in which GJIC was restored by transfection with the connexin 43 (Cx43) gene. When HeLa cells with (Cx43+) or without Cx43 gene (Cx43-) were treated with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) or methylnitrosourea, the Cx43+ cells survived better than Cx43- cells. The mutation frequency at CA repeats was measured with a shuttle vector; in the vector, the coding region of the beta-galactosidase gene was rendered out of frame by insertion of CA repeats, and the frame could be restored by insertion or deletion mutations of the CA repeats. The mutation frequency at CA repeats was 2-fold lower in Cx43+ cells than in Cx43-, both before and after exposure to MNNG or methylnitrosourea (P < 0.05). The frequency of spontaneous HPRT gene mutations, selected by their resistance to 6-thioguanine, was 3-fold lower in Cx43+ cells than Cx43- cells. Similarly, the frequency of MNNG-induced HPRT mutations was significantly higher in Cx43- cells (P < 0.001). Similar results were obtained even when the mutant selection process was carried out in the presence of alpha-glycyrrhetinic acid, a long-term inhibitor of GJIC, suggesting that the observed effect is not due to unwanted killing of cells by GJIC-mediated metabolic cooperation. Thus, our data demonstrate that HeLa cells transfected with the Cx43 gene become more resistant to spontaneous as well as chemically induced genetic changes.
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