ABSTRACT
Previously we've described the obtainment of a subpopulation of cancer stem cells from a human colorec- tal carcinoma cell line MIP101. These cells possess elevated clonogenic and tumorigenic capacities. According to our data, depletion of stem compartment in a cancer cell population blocks its tumorigenicity. The current work is dedicated to the comparison of tumorigenic potential between cell populations with enriched or depleted stem compartment. We show that tumor growth following xenografting of enriched stem cell population can be suppressed by intramuscular injections of ganciclovir. Thus, we report a method to obtain a cell population with high Oct4 promoter expression within the MIP101 colorectal carcinoma cell line and to eliminate these cells from the population in vitro as well as in vivo.
Subject(s)
Biomarkers, Tumor/biosynthesis , Colorectal Neoplasms/pathology , Colorectal Neoplasms/prevention & control , Neoplastic Stem Cells/cytology , Neoplastic Stem Cells/drug effects , Octamer Transcription Factor-3/biosynthesis , Animals , Biomarkers, Tumor/genetics , Cell Culture Techniques , Cell Line, Tumor , Cell Proliferation/drug effects , Colorectal Neoplasms/metabolism , Ganciclovir/pharmacology , Ganciclovir/therapeutic use , Genetic Vectors , Humans , Lentivirus/genetics , Mice , Mice, Nude , Neoplastic Stem Cells/metabolism , Neoplastic Stem Cells/pathology , Octamer Transcription Factor-3/genetics , Puromycin/pharmacology , Xenograft Model Antitumor AssaysABSTRACT
In the current work we make an attempt to compare cancer cells of one origin, but differing in the expression of CEA protein, a clinical marker of metastatic carcinomas, presumably one of the key factors in metastatic activity. We have explored the morphology of cell colonies in vitro, expression patterns of epithelial markers, the ability of these cells to form tumors and metastases in vivo, and evaluated their stem compartment with the aid of a suicidal genetic construct sensitive to the embryonic stem cell marker, Oct4.
Subject(s)
Biomarkers, Tumor/genetics , Colorectal Neoplasms/pathology , Gene Expression Regulation, Neoplastic , Liver Neoplasms/secondary , Neoplastic Stem Cells/pathology , Octamer Transcription Factor-3/genetics , Animals , Biomarkers, Tumor/metabolism , Cell Line, Tumor , Clone Cells , Colorectal Neoplasms/genetics , Colorectal Neoplasms/metabolism , Genetic Vectors , HEK293 Cells , Humans , Lentivirus/genetics , Liver Neoplasms/genetics , Liver Neoplasms/metabolism , Mice , Mice, Nude , Neoplasm Invasiveness , Neoplasm Transplantation , Neoplastic Stem Cells/metabolism , Octamer Transcription Factor-3/metabolism , Tumor BurdenABSTRACT
Investigations of transcriptional regulation of Oct4 gene in mouse embryonic stem cells have revealed an important cis-element--the distal enhancer (DE). DE consists of two functionally significant elements--DEa and DEb. Both elements are necessary to complete the DE-mediated expression of Oct4 gene in pluripotent cells. The most likely candidates for the binding site DEb are Oct4 itself in complex with Sox2 protein. It remains unclear which transcriptional proteins bind to the DEa site and what is the mechanism of the co-operation between the DEa and the DEb. Through the use of using the EMSA and chromatographic fractionation of proteins from extracts of mouse embryonic stem cells and mouse tissues, were isolated proteins specifically interacting with the sequence DEa Oct4 gene.
Subject(s)
Embryonic Stem Cells/metabolism , Gene Expression Regulation, Developmental , Octamer Transcription Factor-3/metabolism , SOXB1 Transcription Factors/metabolism , Transcription, Genetic , Animals , Base Sequence , Binding Sites , Brain Chemistry , Embryo, Mammalian , Embryonic Stem Cells/cytology , Mice , Molecular Sequence Data , Octamer Transcription Factor-3/chemistry , Octamer Transcription Factor-3/genetics , Protein Binding , SOXB1 Transcription Factors/genetics , Signal TransductionABSTRACT
In present publication we describe for the first time the obtainment of cancer stem cells from a weakly metastatic human colorectal carcinoma cell line MIP101 via selecting from the native population the cells that express intensively an embryonic stem cell marker, POU5F1 (Oct4). We provide the evidence that these cells possess an elevated clonogenic and tumorigenic potential when compared to the native population, and this correlates to the hypothesis of cancer stem cells' primary role in the development of malignant neoplasms.
