Method of detecting oncogenesis of hematopoietic cells
Disclosed in a method of detecting cancer using IL-27 receptors. IL27R is a cytokine receptor identified as a novel oncogene from an acute myeloid leukemia patient. It induces cancer-like properties when expressed in cells and can activate a protein that causes various myeloid cell disorders. The data show cytokine receptors play unappreciated roles in mediating activation of signaling pathways in circulatory system cancers. Also method of screening for novel oncogenes using a functional, approach is disclosed using cytokine-dependent cells to screen for transforming events.
1. A method of detecting leukemia, myeloproliferative diseases, myeloid tumors, and acute myeloid leukemia comprising the steps of:
providing a hematopoietic cell line or a sample comprising hematopoietic cells;
contacting the hematopoietic cell line or a sample comprising hematopoietic cells with an antibody which binds a biomarker for the cancerous disorders, wherein the biomarker is related to a component of interleukin 27 receptor;
allowing the biomarker to bind to the antibody; and
detecting the presence of the biomarker in said cell line or said sample using an immunoprotein assay;
wherein the presence of the biomarker is indicative of leukemia, myeloproliferative diseases, myeloid tumors, and acute myeloid leukemia.
2. The method of claim 1 , wherein the biomarker is selected from the group consisting of protein, peptide, or proteineaceous aggregate.
3. The method of claim 2 , wherein the biomarker is WSX-1.
4. The method of claim 1 , wherein the immunoprotein assay is selected from the group consisting of FACS, immunohistochemistry, and Western blot.
5. The method of claim 1 wherein the hematopoietic cell line or a sample comprising hematopoietic cells is mammalian.
6. The method of claim 5 wherein the hematopoietic cell line or a sample comprising hematopoietic cells is human.
7. A method of constructing a cancer research cell line, comprising the steps of
isolating RNA from cells obtained from a patient;
constructing cDNA from the RNA, wherein the cDNA encodes a heterodimeric transmembrane receptor protein for IL-27;
ligating the cDNA into a retroviral vector;
introducing the cDNA into a IL-27 dependent cell culture; and
removing an IL-27 cytokine from cell culture.
8. The method of claim 7 , wherein the gene introduced into the cell culture is a stably transfected into the cells.
9. The method of claim 7 , wherein the cDNA encodes WSX-1.
10. The method of claim 7 , wherein the cytokine dependent cells are selected from the group consisting of 32D and BaF3.
11. The method of claim 7 , further comprising introducing mutant JAK2V617F into the cells.