ABSTRACT
Among the substances isolated from Cryptocarya sp, some styrylpyrones, such as goniothalamin, demonstrate antiproliferative activity in a broad range of human cell lines. In the present study, we assessed the cytotoxicity of a styrylpyrone (cryptomoschatone D2), isolated from Cryptocarya mandiocanna, in HPV-infected (HeLa and SiHa) and uninfected (C33A) human cervical carcinoma cell lines and a human lung fibroblast line (MRC-5). The cytotoxicity was tested by the MTT assay. In this assay, cells were treated with cryptomoschatone D2 at 15, 30, 60 or 90 ?M for 6, 24 or 48 hours, as well as for 6 hours followed by a post-treatment recovery period of 24, 48 or 72 hours. High cytotoxicity (dose- and timedependent) was observed in HeLa, SiHa, C33A and MRC-5 cell lines. Although in general the styrylpyrone cytotoxicity was not significantly different among the cell lines tested, it was apparently stronger in HeLa and C33A than in MRC-5 and SiHa in the 24 or 48-hour treatments. Moreover, HeLa and SiHa were able to recover their ability to proliferate, in direct proportion to the post-treatment recovery time. On the other hand, C33A did not demonstrate a similar post-treatment recovery. We can conclude that cryptomoschatone D2 possesses high dose-dependent or time-dependent cytotoxicity.
Dentre as substâncias isoladas de Cryptocarya sp, algumas estirilpironas, como a goniotalamina, apresentam atividade antiproliferativa em diferentes linhagens celulares. No presente estudo, foram avaliadas as atividades citotóxica de uma estirilpirona (criptomoscatona D2) isolada de Cryptocarya mandiocanna, em linhagens celulares de carcinoma cervical humano infectada por HPV (HeLa e SiHa), não infectada (C33A) e fibroblasto pulmonar humano (MRC-5). A atividade citotóxica foi avaliada pelo ensaio do MTT. No ensaio do MTT, as células foram tratadas com criptomoscatona D2 em 15, 30, 60 e 90 ?M por 6, 24 e 48 horas e por 6 horas com período de recuperação de 24, 48 e 72 horas pós-tratamento. O tratamento com a estirilpirona (criptomoscatona D2) ocasionou elevada citotoxicidade dose-resposta e tempo-resposta em HeLa, SiHa, C33A e MRC-5. Embora não haja diferença estatisticamente significativa de citotoxicidade entre as linhagens, aparentemente a citotoxicidade foi maior em HeLa e C33A (tratamento de 24 e 48 horas) que em MRC-5 e SiHa. Ainda, no período de recuperação, HeLa e SiHa aparentemente restabelecem sua capacidade proliferativa, que é diretamente proporcional ao tempo de recuperação, enquanto o mesmo comportamento não é observado em C33A. Estes resultados sugerem que criptomoscatona D2 possui elevada atividade antiproliferativa dose-resposta ou o tempo resposta.
Subject(s)
Humans , Cryptocarya/toxicity , Neoplasms , Cell Line, Tumor , HeLa CellsABSTRACT
Neuroblastoma, the most common extracranial tumor in childhood, has a wide spectrum of clinical and biological features. The loss of heterozygosity within the 9p21 region has been reported as a prognostic factor. Two tumor suppressor genes located in this region, the CDKN2B/p15 and CDKN2A/p16 (cyclin-dependent kinase inhibitors 2B and 2A, respectively) genes, play a critical role in cell cycle progression and are considered to be targets for tumor inactivation. We analyzed CDKN2B/p15 and CDKN2A/p16 gene alterations in 11 patients, who ranged in age from 4 months to 13 years (male/female ratio was 1.2:1). The most frequent stage of the tumor was stage IV (50 percent), followed by stages II and III (20 percent) and stage I (10 percent). The samples were submitted to the multiplex PCR technique for homozygous deletion analysis and to single-strand conformation polymorphism and nucleotide sequencing for mutation analysis. All exons of both genes were analyzed, but no deletion was detected. One sample exhibited shift mobility specific for exon 2 in the CDKN2B/p15 gene, not confirmed by DNA sequencing. Homozygous deletions and mutations are not involved in the inactivation mechanism of the CDKN2B/p15 and CDKN2A/p16 genes in neuroblastoma; however, these two abnormalities do not exclude other inactivation pathways. Recent evidence has shown that the expression of these genes is altered in this disease. Therefore, other mechanisms of inactivation, such as methylation of promoter region and unproperly function of proteins, may be considered in order to estimate the real contribution of these genes to neuroblastoma genesis or disease progression.