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Disruption of gap junctional intercellular communication in human renal cancer cell lines
M Noguchi1, K Nomata, J I Watanabe
1Department of Urology, Nagasaki University School of Medicine, Japan.
Objectives:
Gap junctional intercellular communication (GJIC) is believed to play an important role in the maintenance of cell homeostasis, and its disruption may be associated with carcinogenesis. However, GJIC has not been detected in many human cancers. We therefore studied the regulation of GJIC in human renal cancer cell lines.
Methods:
We examined the human renal cancer cell lines, ACHN and NT, as well as Madin-Darby canine kidney (MDCK) cells as a positive control, using GJIC assays, Northern blotting to detect connexin 43 mRNA, immunofluorescent staining, and Western blotting of connexin 43 protein.
Results:
GJIC of ACHN and NT was completely blocked. In ACHN cells, connexin 43 mRNA was not altered. However, connexin 43 protein was aberrantly localized and phosphorylated connexin 43 protein had disappeared. Both connexin 43 protein and its mRNA were undetectable in NT cells.
Conclusions:
GJIC in human renal cancer cell lines is impaired and various pathways may inhibit this mechanism in renal cancer. We believe that connexin plays an important role in renal carcinogenesis.
Insights
Gap junctional intercellular communication (GJIC) is impaired in human renal cancer cell lines. Connexin 43 protein alterations and absence suggest its role in renal carcinogenesis.
Area of Science:
- Oncology
- Cell Biology
- Molecular Biology
Background:
- Gap junctional intercellular communication (GJIC) is vital for cell homeostasis.
- Disrupted GJIC is linked to carcinogenesis.
- GJIC is often absent in human cancers, necessitating further study.
Purpose of the Study:
- Investigate the regulation of GJIC in human renal cancer.
- Determine the role of connexin 43 in renal cancer cell lines.
- Identify potential inhibitory pathways of GJIC in renal cancer.
Main Methods:
- Utilized human renal cancer cell lines (ACHN, NT) and MDCK cells.
- Performed GJIC assays.
- Analyzed connexin 43 expression and localization via Northern blotting, immunofluorescence, and Western blotting.
Main Results:
- GJIC was completely blocked in ACHN and NT cells.
- ACHN cells showed altered connexin 43 protein localization and loss of phosphorylated protein, with normal mRNA levels.
- NT cells lacked both connexin 43 protein and mRNA.
Conclusions:
- GJIC is impaired in human renal cancer cell lines.
- Connexin 43 dysregulation and absence contribute to impaired GJIC.
- Connexin 43 plays a significant role in renal carcinogenesis.