fenretinide and parthenolide

Fenretinide (N-4-hydroxyphenyl retinamide, 4HPR) is a synthetic anticancer retinoid that is a well-known apoptosis-inducing agent. Recently, we observed that the apoptosis induced by fenretinide could be effectively enhanced in hepatoma cells by a concomitant treatment with parthenolide, which is a known inhibitor of nuclear factor-kappaB (NF-kappaB). Furthermore, treatment with fenretinide triggered the activation of NF-kappaB during apoptosis, which could be substantially inhibited by parthenolide, suggesting that NF-kappaB activation during fenretinide-induced apoptosis has an antiapoptotic effect. This study investigated the molecular mechanism of this apoptotic potentiation by NF-kappaB inhibition. The genes involved in the enhanced fenretinide-induced apoptosis by parthenolide were identified using the differential display-PCR method and subsequent Northern blot or semiquantitative reverse transcriptase PCR analysis. This study identified 35 apoptosis-related genes including 12 unknown genes that were either up- or down-regulated by parthenolide. Interestingly, one up-regulated gene (HA1A2) was isolated and cloned from the liver cDNA, and was found to be identical to ANKRD1, which is also referred to as the CARP gene. Compared with controls treated with an empty vector or with antisense cDNA, the ectopic expression of ANKRD1 led to reduced colony formation and to enhanced apoptotic cell death in hepatoma cells. These results suggest that ANKRD1 and the other genes, whose expressions were substantially modulated by the parthenolide-mediated inhibition of NF-kappaB activation, play roles in the enhanced drug-induced apoptosis. In addition, this study suggests that those identified genes may be useful in anticancer strategies against hepatoma.

Topics: Amino Acid Sequence; Antineoplastic Combined Chemotherapy Protocols; Apoptosis; Base Sequence; Carcinoma, Hepatocellular; Cell Line, Tumor; Down-Regulation; Drug Synergism; Fenretinide; Gene Expression Regulation, Neoplastic; Humans; Liver Neoplasms; Molecular Sequence Data; Muscle Proteins; NF-kappa B; Nuclear Proteins; Polymerase Chain Reaction; Repressor Proteins; RNA, Messenger; Sesquiterpenes; Up-Regulation

The anticancer effects of N-(4-hydroxyphenyl)retinamide (4HPR), a potential chemopreventive or chemotherapeutic retinamide, are thought to be derived from its ability to induce apoptosis. However, the mechanism of apoptosis induced by 4HPR remains unclear. Thus, this study was designed to identify the gene(s) responsible for induction of apoptosis by 4HPR. Apoptosis was effectively induced by 4HPR in human hepatoma cells. Using the differential display-PCR method, a gene involved in the response to 4HPR was identified, and cells in which the expression of that gene was modulated were analyzed for survival, induction of apoptosis, and cell cycle. GADD153, a gene involved in growth arrest and apoptosis, was preferentially expressed in human hepatoma cells as well as in other cancer cells during 4HPR-induced apoptosis. 4HPR regulates GADD153 expression at the post-transcriptional level in Hep 3B cells and at the transcriptional and post-transcriptional levels in SK-HEP-1 cells, when assayed by in vitro transfection and mRNA stability experiments. To determine the role of the GADD153 protein overexpression that is induced by 4HPR, Hep 3B cells with ectopic overexpression of GADD153 were found to be growth-arrested (at G(1)) and readily underwent apoptosis following treatment with 4HPR or even when they reached confluence. N-Acetyl-l-cysteine or GADD153 antisense significantly protected the cells from 4HPR-induced apoptosis, accompanying by the inhibition of GADD153 overexpression. Parthenolide-mediated overexpression of GADD153 resulted in enhanced 4HPR-induced apoptosis. These results suggest that GADD153 overexpression induced by 4HPR may contribute to the anticancer effects (induction of apoptosis and growth arrest) of 4HPR on cancer cells.

Topics: Acetylcysteine; Antineoplastic Agents; Apoptosis; Carcinoma, Hepatocellular; CCAAT-Enhancer-Binding Proteins; Cell Cycle; DNA Fragmentation; Fenretinide; Flow Cytometry; Gene Expression Profiling; Gene Expression Regulation; Genes, Reporter; Humans; Neoplasms; NF-kappa B; Sesquiterpenes; Transcription Factor CHOP; Transcription Factors; Transcription, Genetic; Tumor Cells, Cultured