Expression of Genes Involved in Porphyrin Biosynthesis Pathway in the Human Renal Cell Carcinoma

Autor(es)

da Rocha Filho, Hugo Nobrega ^{[1, 2]} ; da Silva, Evelin Caroline ^[2] ; Silva, Flavia R. O. ^[3] ; Courrol, Lilia Coronato ^[4] ; de Mesquita, Carlos Henrique ^[5] ; Bellini, Maria Helena ^{[1, 2]}

Afiliação do(s) autor(es)

^[1] Univ Fed Sao Paulo, Dept Med, Disciplina Nefrol, Sao Paulo - Brazil

^[2] IPEN CNEN SP, Ctr Biotecnol, Sao Paulo - Brazil

^[3] IPEN CNEN SP, Ctr Ciencia & Tecnol Mat, Sao Paulo - Brazil

^[4] Univ Fed Sao Paulo, Dept Ciencias Exatas & Terra, Sao Paulo - Brazil

^[5] IPEN CNEN SP, Ctr Tecnol Radiacoes, Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Journal of Fluorescence; v. 25, n. 5, p. 1363-1369, SEP 2015.
• Citações Web of Science: 2

Resumo

Renal cell carcinoma (RCC) remains one of the greatest challenges of urological oncology and is the third leading cause of death in genitourinary cancers. Surgery may be curative when patients present with localized disease. Our previous results demonstrated the autofluorescence of blood PpIX in primary RCC mouse model and an increase in fluorescence intensity as a function of growth of the subcutaneous tumor mass. In another work, a nice correlation between the growth of the tumor mass and tissue fluorescence intensity was found. The aim of this study was to evaluate the expression profile of porphyrin biosynthesis pathway-related genes of human kidney cells. We used two kidney cell lines, one normal (HK2) and another malignant (Caki-1). Endogenous and 5-aminolevolinic acid (ALA) induced protoporphyrin IX (PpIX) HK2 and Caki-1 cells were analyzed by fluorescence spectroscopy. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to measure mRNA of those genes. Emission spectra were obtained by exciting the samples at 405 nm. For ALA untreated cells the maximum fluorescence intensity was detected at 635 nm. The mean peak area of emission spectra in both cells types increased linearly in function of cell number. Besides, basal levels of PpIX autofluorescence of each cell concentration of HK2 samples were significantly lower than those of Caki-1 samples. For ALA-treated cells the mean PpIX spectra shows PpIX emission peak at 635 nm with a shoulder at 700 nm. Analysis of PpIX fluorescence intensity ratio between tumor cells and HK2 cells showed that fluorescence intensity was, on average, 26 times greater in tumor cells than in healthy cells. qRT-PCR revealed that in Caki-1 ALA-treated cells, PEPT gene was significantly up-regulated and FECH and HO-1 genes were significantly down regulated in comparison with HK2 ALA-treated cells. In conclusion, our results demonstrate the preferential accumulation of ALA-induced PpIX in human RCC and also indicate that PEPT1, FECH and HO-1 genes are major contributors to this accumulation. (AU)

• Processo FAPESP: 10/52180-4 - Fisiopatologia e regeneração das lesões renais
• Beneficiário: Nestor Schor
• Modalidade de apoio: Auxílio à Pesquisa - Temático