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Updated: Jul 17, 2026

Detection of Nuclear Blebbing and DNA Leakage in Mammalian Cells by Immunofluorescence
Published on: January 17, 2025
NF-kappaB2 processing and p52 nuclear accumulation after androgenic stimulation of LNCaP prostate cancer cells
Laurent Lessard1, Fred Saad, Cécile Le Page
1Université de Montréal, Centre de recherche du CHUM et Institut du cancer de Montréal, 1560 Sherbrooke East, Montréal, QC, Canada, H2L 4M1.
Abstract:
Several reports suggest that androgen signalling interferes with canonical RelA-p50 activity in androgen-sensitive cells. Whether this also occurs with non-canonical NF-kappaB subunits has not been studied. Here we report that androgenic stimulation of LNCaP cells with the androgen analogue R1881 appears to positively regulate the non-canonical NF-kappaB pathway as p52 accumulates both in the cytoplasm and nucleus after 48-72 h of stimulation. In contrast to TNF-alpha stimulation, androgen stimulation fails to induce RelB expression and is absent from nucleus of R1881-treated LNCaP cells. Electromobility shift assays reveal a time-dependent change in the nature of NF-kappaB complexes actively bound to DNA after 72 h of androgenic stimulation concomitant with the appearance of p52-containing complexes. Co-immunoprecipitation studies indicate that newly produced p52 can exist as a heterodimer with RelA or p50, but may be mainly present as a homodimer. RNAi experiments targeting IKK-alpha and IKK-beta show that the R1881-induced nuclear accumulation of p52 is IKK-alpha-dependent. These results point to a novel mechanism by which androgens regulate NF-kappaB and provide a rationale for further studies into the biological significance of non-canonical NF-kappaB signalling in prostate cancer.
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