EFFECT OF BRCA1 AFTER UV-B-IRRADIATION IN CELL CYCLE PROGRESSION BETWEEN HUMAN MELANOMA CELL LINES DETERMINED BY FLOW CYTOMETER.

Authors

  • Moayed Naji Majeed College of medicine ,dep.of pediatrics,University of Thi-Qar
  • Ahmed A.Naser Al-Amiry College of medicine ,dep.of physiology,University of Thi-Qar
  • Francesco S. Costanzo

Abstract

The aim of the current study was to evaluated the effect of BRCA1(breast cancer 1 early-0nsent) after UV-B-irradiation in a panel of human malignant melanoma cells using flow cytometry method. Different human melanoma cell lines (SK-MEL28,SK-MEL93, ,SK-MeL93/shBRCA1 and SK-Mel93/shplKO.1) were cultured for triplicaten experiments. Cells were  exposed to UV-B (10mJ/cm2)for different point time(1/2,1,3,6,9,12,and 24h). Cells at 0hr were a non-irradiation control. The cell cycle was evaluated by flow cytometry. SK-MEL28 cell line  seemed to be synchronized at the G1/S boundary phase until 12h after UVB-irradiation. a decrease in the fraction of S cells and a reciprocal increase of cells in Sub-G1 phase was observed , from 12 to 24h.While, the SK-MEL93 a rapid increase (about 35%) of cells in Sub-G1 phase was observed already 6h after UVB irradiation. Moreover, we found that the  SKMel93/shBRCA1 cells seemed an increase (about 15%) of cells in Sub-G1 phase already 12h after UVB irradiation compared with cells infected with the empty lentiviral vector (SK-Mel93/shplKO.1). we conclude that an important role played by BRCA1 providing new insights into the molecular mechanisms underlying UV-induced melanomagenesis.  

References

Lomas J, Martin-Dugue P,pons M, & Quintanilla M. The genetics of malignant melanoma.Biosci. 2008; 13(1): 5071-5093.

Tucker M.A.& Goldstein A.M. Melanoma etiology: Where are we? Oncogene. 2003;22:3042-3052.

Duncan, I.M., et al. Metastatine expression and prognosis in cutaneous malignant melanoma.J Clin. Oncol. 2001, 19: 568-576.

Sabatino M, Stroncek DF, Klein H, Marincola FM.& Wang E. Stem cell in melanoma development. Cancer lett.2009,279(2): 119-125.

Pollock P. M.,Yu F., Qiu L., Parsons P. G.,& Hayward N. K.K. Evidence for UV induction of CDKN2 mutations in melanoma cell line. Oncogene. 1995; 11:663-668.

Peris K., Chimeti S., Fargnoli M. C., Valeri P., Kerl H., & Wolf P. UV fingerpreint CDKN2a but no p14ARF mutations in sporadic melanomas. J Invest . Dermatol. 1999;112:825-826.

Hussian, C.J.,et al. Germline p16 mutations. Nat.Genet .1994.8:15-21.

Ortonne Jp:From actinic keratosis to squamous cell carcinoma. Br J Dermatol. 2002;146 (61):20-23.

D E Brash, J A Rudolph, J A Simon, A Lin, G J McKenna, H P Baden, A J Halperin, and J Ponten.: A role for sunlight in skin cancer: UV-induced p53 mutations in squamous cell carcinoma. Proc Natl Acad Sci U S A. 1991; 88(22): 10124–10128.

Ziengler AD. et al: Mutation hotsipots due sunlight in the p53 gene of melanoma skin cancer. Proc. Nqatl Acad Sci USA 1993;90:4216-4220.

Halmbach H, Rossman E, Korn M, Schadendorf D. Drug resistance in Human melanoma. Int J Cancer 2001;93:617-22

Joiner MC, Van Der Kogel AJ. The linear quadratic approach to fractionation and calculation of isoeffect relationships . In Stal GG, Editor. Basic Clinical Radiobiology. London; 1997. p107-21

Downloads

Published

2021-05-21

Issue

Section

Articles