The effect of differentiating and apoptotic agents on notch signalling pathway in hepatoblastoma.

BACKGROUND/AIMS: Notch expression is not yet determined in hepatoblastoma. In this study the effect of chemotherapeutics (cisplatin, doxorubicin, cytosin arabinoside); differentiating agent (13 cis-retinoic acid) and apoptotic agents (5-aza-2'-deoxycytidine, arsenic trioxide) on notch expression in hepatoblastoma were evaluated.

METHODOLOGY: After HepG2 cell line was cultured and the agents and their combinations were applied for 24 hour in pre-optimized 50% lethal doses, RNA isolation and cDNA converting, expression of 84 custom array genes of notch signaling pathway (SABiosciences, PAT059F-24) was determined by Real Time PCR. The methylation status of 6 genes that showed more than 5 fold changes compared with control group were explored by Methylation qPCR Assay. High expressed genes are HDAC1, NFKB1, CHUK, CDKN1A, and CBL. Low expressed genes are DLL1, CD44, FZD2, GLI1, IL17B, LMO2, NOTCH1, LOR, PAX5, PT-CRA, SH2D1A, and WISP1. The genes searched for methylation (DLL1, HEY1, DTX1, HDAC1, NOTCH2 and JAG1) were not found to be related with methylation.

RESULTS: The high expressed genes are related with cell proliferation. The main signaling genes that are closed to notch in signaling pathway are low expressed in hepatoblastoma. The agents do not show prominent effect of gene expression in many genes and methylation is not the reason of expression changes. The use of retinoic acid in the control of minimal residual disease of hepatoblastoma should be discussed. 5 aza "cytidin" the demethylating agent is not advised in treatment according to our results.

CONCLUSION: Cisplatin as main chemotherapeutic agent treatment is shown to change gene expression levels in notch signalling pathway in hepatoblastoma.