Cisplatin-induced premature senescence with concomitant reduction of gap junctions in human fibroblasts.
To examine the role of gap junctions in cell senescence, the changes of gap junctions in cisplatin-induced premature senescence of primary cultured fibroblasts were studied and compared with the replicative senescent human fibroblasts. Dye transfer assay for gap junction function and immunofluorescent staining for connexin 43 protein distribution were done respectively. Furthermore, cytofluorimetry and DAPI fluorescence staining were performed for cell cycle and apoptosis analysis. p53 gene expression level was detected with indirect immunofluorescence. We found that cisplatin (10 mM) treatment could block cell growth cycle at G1 and induced premature senescence. The premature senescence changes included high frequency of apoptosis, elevation of p53 expression, loss of membranous gap junctions and reduction of dye-transfer capacity. These changes were comparable to the changes of replicative senescence of human fibroblasts. It was also concluded that cisplatin could induce premature senescence concomitant with inhibition of gap junctions in the fibroblasts. Loss of functional gap junctions from the cell membrane may account for the reduced intercellular communication in the premature senescent fibroblasts. The cell system we used may provide a model useful for the study of the gap junction thus promoting agents against premature senescence.
Zhao
Wei
W
Peking University, School of Oncology, and Beijing Institute for Cancer Research, Beijing 100034, China.
Lin
Zhong Xiang
ZX
Zhang
Zhi Qian
ZQ
D017209:Apoptosis / Q000187:drug effects; D001692:Biological Transport / Q000187:drug effects; D002450:Cell Communication / Q000187:drug effects; D002453:Cell Cycle / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002462:Cell Membrane / Q000737:chemistry / Q000187:drug effects; D002478:Cells, Cultured; D016922:Cellular Senescence / Q000187:drug effects*; D002945:Cisplatin / Q000494:pharmacology*; D018031:Connexin 43 / Q000032:analysis / Q000378:metabolism; D005347:Fibroblasts / Q000737:chemistry / Q000166:cytology / Q000187:drug effects*; D005434:Flow Cytometry; D017629:Gap Junctions / Q000187:drug effects*; D006801:Humans; D007150:Immunohistochemistry; D007211:Indoles / Q000737:chemistry; D007546:Isoquinolines / Q000494:pharmacology; D008856:Microscopy, Fluorescence; D008938:Mitosis / Q000187:drug effects; D016159:Tumor Suppressor Protein p53 / Q000032:analysis / Q000378:metabolism
D018031:Connexin 43; D007211:Indoles; D007546:Isoquinolines; D016159:Tumor Suppressor Protein p53; C007293:DAPI; C017475:lucifer yellow; D002945:Cisplatin