Comparison of proliferation and genomic instability responses to WRN silencing in hematopoietic HL60 and TK6 cells.
BACKGROUND
Werner syndrome (WS) results from defects in the RecQ helicase (WRN) and is characterized by premature aging and accelerated tumorigenesis. Contradictorily, WRN deficient human fibroblasts derived from WS patients show a characteristically slower cell proliferation rate, as do primary fibroblasts and human cancer cell lines with WRN depletion. Previous studies reported that WRN silencing in combination with deficiency in other genes led to significantly accelerated cellular proliferation and tumorigenesis. The aim of the present study was to examine the effects of silencing WRN in p53 deficient HL60 and p53 wild-type TK6 hematopoietic cells, in order to further the understanding of WRN-associated tumorigenesis.


RESULTS
We found that silencing WRN accelerated the proliferation of HL60 cells and decreased the cell growth rate of TK6 cells. Loss of WRN increased DNA damage in both cell types as measured by COMET assay, but elicited different responses in each cell line. In HL60 cells, but not in TK6 cells, the loss of WRN led to significant increases in levels of phosphorylated RB and numbers of cells progressing from G1 phase to S phase as shown by cell cycle analysis. Moreover, WRN depletion in HL60 cells led to the hyper-activation of homologous recombination repair via up-regulation of RAD51 and BLM protein levels. This resulted in DNA damage disrepair, apparent by the increased frequencies of both spontaneous and chemically induced structural chromosomal aberrations and sister chromatid exchanges.


CONCLUSIONS
Together, our data suggest that the effects of WRN silencing on cell proliferation and genomic instability are modulated probably by other genetic factors, including p53, which might play a role in the carcinogenesis induced by WRN deficiency.
Genes and Environment Laboratory, Division of Environmental Health Sciences, School of Public Health, University of California, Berkeley, California, United States of America.
Xuefeng
X
Ren
Sophia
S
Lim
Zhiying
Z
Ji
Jessica
J
Yuh
Vivian
V
Peng
Martyn T
MT
Smith
Luoping
L
Zhang
D002453:Cell Cycle; D002460:Cell Line; D049109:Cell Proliferation; D004249:DNA Damage; D005090:Exodeoxyribonucleases / Q000172:deficiency / Q000235:genetics*; D020868:Gene Silencing; D042822:Genomic Instability; D018922:HL-60 Cells; D006801:Humans; D053484:RecQ Helicases / Q000172:deficiency / Q000235:genetics*; D016159:Tumor Suppressor Protein p53; D014898:Werner Syndrome; D000071657:Werner Syndrome Helicase
D016159:Tumor Suppressor Protein p53; D005090:Exodeoxyribonucleases; D053484:RecQ Helicases; C098785:WRN protein, human; D000071657:Werner Syndrome Helicase
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