Nuclear translocation of ADAM-10 contributes to the pathogenesis and progression of human prostate cancer - PubMed (original) (raw)

. 2007 Nov;98(11):1720-6.

doi: 10.1111/j.1349-7006.2007.00601.x. Epub 2007 Aug 28.

Hideki Enokida, Hiroyuki Kubo, Ichiro Kagara, Ryouichirou Matsuda, Kazuki Toki, Hiroaki Nishimura, Takeshi Chiyomaru, Shuichi Tatarano, Toshihiko Idesako, Kenryu Nishiyama, Masayuki Nakagawa

Affiliations

Nuclear translocation of ADAM-10 contributes to the pathogenesis and progression of human prostate cancer

Takashi Arima et al. Cancer Sci. 2007 Nov.

Abstract

A disintegrin and metalloproteases (ADAM) are cell membrane-anchored proteins with potential implications for the metastasis of human cancer cells via cell adhesion and protease activities. In prostate cancer (PC), the ADAM-10 protein showed a nuclear localization whereas in benign prostate hypertrophy (BPH) it was predominantly bound to the cell membrane. We hypothesized that the pathogenesis and progression of PC are attributable to the nuclear translocation of ADAM-10. Immunoblotting revealed that after 5alpha-dihydrotestosterone treatment, a 60-kDa active form of ADAM-10 was increased in the nuclear fraction but decreased in the cell membrane and cytoplasmic fractions of human androgen-dependent PC cells. Immunocytochemistry revealed that after 5alpha-dihydrotestosterone treatment, the ADAM-10 protein was translocated from the cell membrane to the nucleus. Coimmunoprecipitation of androgen receptor and ADAM-10 was detected in the nuclear fraction but not in the cell membrane and cytoplasmic fractions. Immunohistochemical study of 64 PC and 20 BPH samples showed that the intensity of ADAM-10 staining was significantly higher in the nuclei of PC cells than in the nuclei of BPH cells (P < 0.0001). It was also significantly lower in the cell membrane of PC cells than in the cell membrane of BPH cells (P = 0.0017). Nuclear staining intensity was significantly correlated with the clinical T-factor (P = 0.004), the Gleason score (P < 0.0001) and preoperative prostate-specific antigen levels (P = 0.0061). ADAM-10 small interfering RNA transfectants showed a significant decrease in cell growth compared to the controls. Our results suggest that in human PC, the nuclear translocation of ADAM-10 coupled with the androgen receptor is involved in tumor growth and progression.

PubMed Disclaimer

Figures

Figure 1

Figure 1

Cellular localization of a disintegrin and metalloprotease (ADAM)‐10 protein in LNCaP cells treated with different concentrations of 5α‐dihydrotestosterone (DHT). (a) ADAM‐10 expression: the pro‐form (~100 kDa), intermediate form (~80 kDa) and active form (~60 kDa), detected by immunoblotting in whole‐cell lysates. Densitometric analysis revealed that there was no difference in the level of active ADAM‐10 expression between whole‐cell lysates from cells exposed to different concentration of DHT. (b) ADAM‐10 expression detected by immunoblotting in the cell membrane and cytoplasm fractions and the nuclear factions. At higher DHT concentrations, the membrane and cytoplasmic fractions manifested a significant decrease, and the nuclear fractions showed a significant increase in active ADAM‐10 expression. Glyceraldehyde‐3‐phosphate dehydrogenase and octamer‐binding transcription factor‐1 were the markers for the cytoplasmic and nuclear fractions, respectively. (c) ADAM‐10 protein expression visualized by immunocytochemistry. In DHT‐treated cells, ADAM‐10 protein translocated from the cell membrane to the nucleus. AU, arbitrary units. Scale bar = 20 µm.

Figure 2

Figure 2

Effect of small interfering RNA (siRNA) a disintegrin and metalloprotease (ADAM)‐10 treatment on the growth of LNCaP cells. (a) ADAM‐10 mRNA expression determined by reverse transcription–polymerase chain reaction assay. Lane 1, untreated (parental) cells; lane 2, cells treated with transfection reagent only (mock); lane 3, cells treated with non‐silencing siRNA (si‐control); lane 4, cells treated with si‐ADAM‐10 (si‐ADAM‐10). (b) Cell growth determined by MTT assay. Compared to the controls, cell growth was significantly reduced by exposure to si‐ADAM‐10. In the absence of 5α‐dihydrotestosterone (DHT) treatment, the difference on day 4 was P < 0.01. In the presence of DHT treatment, it was P < 0.01 (day 3) and P < 0.0001 (day 4).

Figure 3

Figure 3

Androgen receptor (AR) interacts with ADAM‐10. Anti‐ADAM‐10 antibody was used to immunoprecipitate ADAM‐10 and associated proteins. Immunoblotting was with anti‐AR‐ and anti‐ADAM‐10 antibody. ADAM‐10 was associated with AR in the nuclear factions but not in the cell membrane and cytoplasmic fractions of LNCaP cells. By densitometric analysis, co‐immunoprecipitated AR protein in the nuclear fractions tended to increase with the dose of 5α‐dihydrotestosterone (P = 0.09). The presence of ADAM‐10 protein expression in all lanes confirmed successful immunoprecipitation. The results are representative of three independent experiments.

Figure 4

Figure 4

The expression of a disintegrin and metalloprotease (ADAM)‐10 in benign prostate hypertrophy (BPH) and prostate cancer (PC) cells. (a) Representative immunohistochemical staining results for ADAM‐10. The large insert in (a) is a magnification of the small boxed area (magnification ×200 and ×400). (b) ADAM‐10 expression scores in the cell membrane and nuclei of BPH and PC cells. The score for cell membrane staining was significantly higher in BPH than PC (P = 0.0017). Conversely, the H‐score for nuclear staining intensity was significantly lower in BPH than PC (P < 0.0001).

Figure 5

Figure 5

Correlation between a disintegrin and metalloprotease (ADAM)‐10 expression and clinicopathological features in human prostate cancer (PC). (a) Representative results of immunohistochemical staining for ADAM‐10 in low‐ and high‐grade PC. The large inserts in (a) are a magnification of the small boxed area (magnification ×200 and ×400). (b) Correlation between ADAM‐10 expression in the nucleus and cell membrane of PC cells and clinicopathological features. The H‐score for nuclear staining intensity increased with advanced clinicopathological features. In contrast, the cell membrane staining score was higher in the presence of a lower Gleason score and a lower preoperative prostate‐specific antigen level.

Similar articles

Cited by

References

    1. Huovila AP, Turner AJ, Pelto‐Huikko M, Kärkkäinen I, Ortiz RM. Shedding light on ADAM metalloproteinases. Trends Biochem Sci 2005; 30: 413–22. - PubMed
    1. Seals DF, Courtneidge SA. The ADAMs family of metalloproteases: multidomain proteins with multiple functions. Genes Dev 2003; 17: 7–30. - PubMed
    1. Arribas J, Borroto A. Protein ectodomain shedding. Chem Rev 2002; 102: 4627–38. - PubMed
    1. Nunan J, Small DH. Regulation of APP cleavage by alpha‐, beta‐ and gamma‐secretases. FEBS Lett 2000; 483: 6–10. - PubMed
    1. McGowan PM, Ryan BM, Hill AD, McDermott E, O’Higgins N, Duffy MJ. ADAM‐17 expression in breast cancer correlates with variables of tumor progression. Clin Cancer Res 2007; 13: 2335–43. - PubMed

MeSH terms

Substances

LinkOut - more resources