title: Immunocytochemical studies on the nuclear ubiquitous casein and cyclin-dependent kinases substrate following 5-aminolevulinicacid-mediated photodynamic therapy on MCF-7 cells.
abstract: BACKGROUND
  Recent data indicates that nuclear ubiquitous casein and cyclin-dependent kinases substrate (NUCKS) may play role in tumor growth. In present study authors examined whether photodynamic therapy with 5-aminolevulinic acid (5-ALA) induces NUCKS expression in breast cancer cell line, MCF-7.
  
  
  METHODS
  In the experiment concentration of 5-ALA was 6.5mM. Excitation wavelength was 630 ± 20 nm, total light dose of light 5 or 10 J/cm(2) and irradiance 60 mW/cm(2) was used. Cells were collected at established time points and Western blot and immunocytochemical studies were performed using antibody against NUCKS.
  
  
  RESULTS
  Studies proved strong cytotoxic effects in cells following PDT with 6.5mM of precursor and 10 J/cm(2). Western blot analysis revealed the strongest expression of NUCKS at 7h after PDT. At next time points, 18 and 24h, expression of NUCKS decreased and became similar to that of control group. Further immunocytochemical studies showed very strong expression of NUCKS following PDT with 5-ALA and light irradiation of 5 J/cm(2). Early, at 0 h, that expression was predominantly seen in nuclei, while at 7h expression of NUCKS was observed in disseminated manner within entire cells in both nuclei and cytoplasm, with prevalence of cytoplasmic staining.
  
  
  CONCLUSIONS
  Authors suggest that NUCKS is involved in cellular responses following PDT, and since parallel induction of NUCKS and proapoptotic marker Bax and inhibition of anti-apoptotic Bcl-2 was observed, this protein might also be involved in induction of apoptosis following PDT.
authors.affiliation: Department of Clinical Biochemistry, Wroclaw Medical University, 50-367 Wrocław, Poland.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Hotowy
authors.forename: Marta
authors.initials: M
authors.lastname: Woźniak
authors.forename: Kamila
authors.initials: K
authors.lastname: Duś
authors.forename: Elżbieta
authors.initials: E
authors.lastname: Czapińska
authors.forename: Beata
authors.initials: B
authors.lastname: Osiecka
authors.forename: Małgorzata
authors.initials: M
authors.lastname: Krzystek-Korpacka
authors.forename: Andrzej
authors.initials: A
authors.lastname: Bronowicz
authors.forename: Jacek
authors.initials: J
authors.lastname: Wiśniewski
authors.forename: Andrzej
authors.initials: A
authors.lastname: Gamian
authors.forename: Grzegorz
authors.initials: G
authors.lastname: Terlecki
authors.forename: Piotr
authors.initials: P
authors.lastname: Ziółkowski
mesh_terms: D000622:Aminolevulinic Acid / Q000627:therapeutic use*; D017209:Apoptosis / Q000187:drug effects* / Q000528:radiation effects*; D014408:Biomarkers, Tumor / Q000378:metabolism*; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects; D006801:Humans; D061986:MCF-7 Cells; D009687:Nuclear Proteins / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism*; D010778:Photochemotherapy / Q000379:methods*; D013379:Substrate Specificity
chemical_list: D014408:Biomarkers, Tumor; C426654:NUCKS1 protein, human; D009687:Nuclear Proteins; D010750:Phosphoproteins; D000622:Aminolevulinic Acid
keywords: Aminolevulinic acid; MCF-7 cells; Nuclear ubiquitous casein and cyclin-dependent kinases substrate; Photodynamic therapy