Project description:Cancer stem cells (CSCs) or tumor-initiating cells (TICs) organize a cellular hierarchy in a similar fashion to normal stem cell systems and exhibit high tumorigenic activity in xenograft transplantation assay. Disulfiram (DSF) could preferentially eradicate TICs, but the molecular machinery of its effect against TICs still remains largely unknown. We found that flow cytometric analyses showed that DSF but not 5-FU drastically reduces the number of tumor-initiating HCC cells. We conducted microarray analyses to examine gene expression profiling in DSF-treated tumor-initiating HCC cells. Purified EpCAM-positive HCC cells treated with Disulfiram or 5-FU were subjected to RNA extraction and hybridization on Agilent microarrays. Data were obtained for tripricate samples from three independent experiments.
Project description:Cancer stem cells (CSCs) or tumor-initiating cells (TICs) organize a cellular hierarchy in a similar fashion to normal stem cell systems and exhibit high tumorigenic activity in xenograft transplantation assay. Disulfiram (DSF) could preferentially eradicate TICs, but the molecular machinery of its effect against TICs still remains largely unknown. We found that DSF but not 5-FU erradicates tumor-initiating HCC cells. To compare the gene expression profiles in EpCAM-positive cells and EpCAM-negative cells, we conducted microarray analyses of EpCAM-negative HCC cells treated with DSF or 5-FU. Purified EpCAM-negative HCC cells treated with Disulfiram or 5-FU were subjected to RNA extraction and hybridization on Agilent microarrays.
Project description:Cancer stem cells (CSCs) or tumor-initiating cells (TICs) organize a cellular hierarchy in a similar fashion to normal stem cell systems and exhibit high tumorigenic activity in xenograft transplantation assay. Disulfiram (DSF) could preferentially eradicate TICs, but the molecular machinery of its effect against TICs still remains largely unknown. We found that DSF but not 5-FU erradicates tumor-initiating HCC cells. To compare the gene expression profiles in EpCAM-positive cells and EpCAM-negative cells, we conducted microarray analyses of EpCAM-negative HCC cells treated with DSF or 5-FU.
Project description:Cancer stem cells (CSCs) or tumor-initiating cells (TICs) organize a cellular hierarchy in a similar fashion to normal stem cell systems and exhibit high tumorigenic activity in xenograft transplantation assay. Disulfiram (DSF) could preferentially eradicate TICs, but the molecular machinery of its effect against TICs still remains largely unknown. We found that flow cytometric analyses showed that DSF but not 5-FU drastically reduces the number of tumor-initiating HCC cells. We conducted microarray analyses to examine gene expression profiling in DSF-treated tumor-initiating HCC cells.
Project description:Increased ?-fetoprotein (AFP) levels have been reported to predict a poor prognosis in hepatocellular carcinoma (HCC). We assessed the mechanism of AFP involvement in the progression of HCC and determined whether AFP could be a molecular target. We used human HCC cell lines to assess proliferation and apoptosis response to exogenous AFP. We introduced AFP small interfering RNA (siRNA) into HCC cell lines to examine whether it could inhibit cell proliferation and anti-apoptotic properties. The effects of systemically introduced AFP siRNA were assessed using a tumor xenotransplantation model. The effects of AFP on gene expression in HCC cell lines and human HCC specimens were examined. Exogenous AFP induced cell proliferation dose-dependently and inhibited apoptosis induced by 5-fluorouracil (5-FU) in all cell lines examined. AFP siRNA inhibited the proliferation of AFP-producing HCC cell lines and induced apoptosis in co-cultures with 5-FU. Tumor sizes in mice treated with AFP siRNA were significantly smaller than those in controls. AFP siRNA administration in mice induced a low proliferation index and a high apoptosis index in tumors. cDNA microarray analysis, reverse transcription-polymerase chain reaction (RT-PCR), and Western blot using HepG2 and HLE cells with AFP showed that AFP reduced expression of genes related to apoptosis (DFFB) and tumor suppression (NDRG2). Expression of these molecules was also suppressed in human HCC tissues that overexpress AFP. AFP is not only a passive tumor marker, but also an active tumor stimulator through several mechanisms. AFP siRNA introduction may be of possible therapeutic use for HCC. Keywords: Genetic modification Two-condition experiment, Control vs. AFP-stimulated cells.
Project description:We identify a new cell subset Ter119+CD45- small cells that promotes tumor metastasis in hepatocellular carcinoma (HCC)-bearing mice. We used microarrays to detail the gene expression of Ter119+CD45- cells comparing with CD45- cells in the spleen of hepatocellular carcinoma (HCC)-bearing mice. Ter119+CD45- cells in the spleen of hepatocellular carcinoma (HCC)-bearing mice, being sorted by a MoFlo high-speed cell sorter, were prepared for RNA extraction and hybridization on Affymetrix microarrays. The CD45+ cells from the same tumor bearing mice were prepared as control.
Project description:To investigate the relationship between MIF and SPP1, we induced THP-1 cells with lentivirus carrying SPP1 shRNA and negative control virus into HCC-TAMs and 4IPP-treated HCC-TAMs (tumor associated macrophages in hepatocellular carcinoma).
Project description:Tumor cells were microdissected from FFPE sections of hepatocellular carcinoma (HCC) samples. micro RNA expression were correlated to clinical outcome and sorafenib-therapy. miRNA was labeled with the Affymetrix FlashTag Biotin HSR RNA Labeling Kit 20 microdissected hepatocellular carcinoma samples with detailed clinical data
Project description:The cancer initiating cells (CICs) act as a tumor initiation source. Recent studies have shown that CICs contribute to chemoresistance and radioresistance. The aims of this study were to investigate the relationship of CD133+ liver CICs and radiation resistance and to define a possible mechanism for radioresistance in hepatocellular carcinoma (HCC).
