The purpose of this study was to investigate the expression feature of a human tumor related gene chp2 in leukemia primary cells and leukemia cell lines, real-time quantitative PCR (RQ-PCR) was performed to detect the expression level of chp2 gene in peripheral blood mononuclear cells from 10 healthy individuals (as control) and 24 cases of leukemia, and in 4 kinds of leukemia cell lines. The results showed that the detection rate of chp2 gene in 10 normal controls was 80%, positive expression was (0.744 +/- 0.682) x 10(5) cps/microl. The expression levels of chp2 mRNA leukemia primary cells and leukemia cell lines were significantly higher than that in the normal control (p < 0.05). The expression levels of chp2 mRNA were higher in AML cells (7 cases), CML cells (6 cases), ALL cells (7 cases) and CLL cells (4 cases), and their expression levels were (11.637 +/- 5.588), (6.122 +/- 3.785), (4.262 +/- 2.561) and (3.434 +/- 1.974) x 10(5) cps/microl respectively. Gene chp2 positively expressed in four kinds of leukemia cell lines, and the expression levels in K562 cells, Jurkat cells, HL-60 cells and M07e cells were (5.243 +/- 1.852), (4.463 +/- 1.621), (4.137 +/- 1.837) and (2.578 +/- 1.137) x 10(6) cps/microl respectively. The expression level in leukemia cell lines was higher than that in primary cells. It is concluded that the human tumor related gene chp2 expression in leukemia primary cells and leukemia cell lines significantly increase, which may play an important role in growth process of leukemia cells.
Calcium-Binding Proteins ; genetics ; metabolism ; HL-60 Cells ; Humans ; Jurkat Cells ; K562 Cells ; Leukemia ; genetics ; metabolism ; pathology ; Neoplasm Proteins ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism

This study was purposed to explore the mechanism of central nervous system (CNS) leukemia resulting from brain metastasis of human acute T-cell leukemia (T-ALL) cells and the role of MIP-1α in migration of Jurkat cells through human brain microvascular endothelial cells (HBMEC). The real-time PCR, siRNA test, transendothelial migration test, endothelial permeability assay and cell adhesion assay were used to detect MIP-1α expression, penetration and migration ability as well as adhesion capability respectively. The results showed that the MIP-1α expression in Jurkat cells was higher than that in normal T cells and CCRF-HSB2, CCRF-CEM , SUP-T1 cells. The MIP-1α secreted from Jurkat cells enhanced the ability of Jurkat cells to penetrate through HBMEC, the ability of Jurkat cells treated by MIP-1α siRNA to adhere to HBMEC and to migrate trans endothelial cells decreased. It is concluded that the MIP-1α secreted from Jurkat cells participates in process of penetrating the Jurkat cells through HBMEC monolayer.
Brain Neoplasms ; pathology ; secondary ; Cell Adhesion ; Cell Movement ; Chemokine CCL3 ; metabolism ; Endothelial Cells ; pathology ; Endothelium, Vascular ; pathology ; Humans ; Jurkat Cells ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; pathology

The study was aimed to explore the correlation of expression of pten mRNA and PTEN protein with AKT phosphorylation levels in various types of leukemia and to elucidate its role in the pathogenesis of leukemia so as to provide some evidence for using PI3K/AKT pathway inhibitors in future. 128 de novo leukemia patients were enrolled in this study, including 61 AML cases, 27 ALL cases, 24 CML cases, and 16 CLL cases. 21 volunteers were selected as normal control. The RT-PCR and Western blot were used to assay the expressions of pten mRNA, PTEN protein, and P-AKT protein in Jurkat cells, bone marrow mononuclear cells of patients respectively. The results showed that the expressions of pten mRNA and PTEN protein in Jurkat cells were lower than that in normal control group; the expression of pten mRNA in AML group was lower than that in normal control group, but the difference was not significant (p=0.274); the expressions of pten mRNA in ALL, CML, CLL each group were lower than that in normal control group, and the difference was significant (p<0.05). Compared with normal control group, the expression of PTEN protein was lower and the expression of P-AKT protein was higher in AML, ALL, CML, CLL each group, the difference were significant (p<0.05). It is concluded that the a lower expression of PTEN protein and higher expression of P-AKT protein may play an important role on leukemia pathogenesis, and inactivation of PTEN protein mainly occurs in the level of protein translation.
Bone Marrow Cells ; metabolism ; Case-Control Studies ; Humans ; Jurkat Cells ; Leukemia ; metabolism ; pathology ; PTEN Phosphohydrolase ; metabolism ; Phosphorylation ; Proto-Oncogene Proteins c-akt ; metabolism ; RNA, Messenger ; genetics

OBJECTIVETo investigate the inhibition effect of leukemic bone marrow stromal cells (BMSCs) on daunorubicin (DNR) induced apoptosis of human Jurkat cell line, and analyze the differentially expressed genes between Jurkat cells cocultured with leukemic BMSCs or without.

METHODSSuppression subtractive hybridization (SSH) was employed to establish subtracted cDNA library of differentially expressed genes in Jurkat cells cocultured with leukemic BMSCs and DNR. The cDNA fragments were sequenced and analyzed.

RESULTSThe differentially expressed gene cDNA library was successfully developed. Primary screening was done by reverse Northern hybridization. Thirty up-regulated and 22 down-regulated cDNA fragments were isolated and sequenced. Analysis and comparison were performed in GenBank using BLAST. These genes are related to cell cycle regulation, cell apoptosis and energy metabolism.

CONCLUSIONLeukemic BMSCs influence gene expression of Jurkat cells. The resulting differentially expressed genes might be associated with the protection of leukemic cells by BMSCs from injury.

Apoptosis ; drug effects ; Bone Marrow Cells ; drug effects ; metabolism ; pathology ; Cells, Cultured ; Coculture Techniques ; Daunorubicin ; pharmacology ; Gene Expression Profiling ; Gene Expression Regulation, Leukemic ; drug effects ; Gene Library ; Humans ; Jurkat Cells ; Stromal Cells ; drug effects ; metabolism ; pathology

OBJECTIVETo detect mutations of human telomeric repeat binding factor 1 (TERF1) gene in 11 malignant hematopoietic cell lines, which have positive telomerase activity, and evaluate the significance of the mutations.

METHODSGenome structure of TERF1 was predicted by using biology information program, and verified by PCR and sequencing. Telomerase activity was detected by telomeric repeat amplification (TRAP)-ELISA. PCR and sequencing were used to detect mutation of each exon of TERF1 in 11 cell lines, including myelogenous leukemia cell lines K562, HL-60, U-937, NB4, THP-1, HEL and Dami; lymphoblastic leukemia cell lines 6T-CEM, Jurkat and Raji and MDS-RAEB cell line MUTZ-1. Five DNA samples from healthy volunteers were detected as normal controls.

RESULTSTERF1 gene has 10 exons and spans 38.6 kb. All the 11 cell lines showed positive telomerase activity. No mutation was found in all exons of TERF1 in the 11 cell lines. However, 4 variants were found in intron1, 2 and 8 near exon1, exon2 and exon9, respectively. The variants in MUTZ-1 was different from those in leukemia cell lines; but no difference was found between the variants in myelogenous and lymphoblastic leukemia cell lines.

CONCLUSIONTERF1 mutation is probably not among the main factors of the gene dysfunction in malignant hematopoietic diseases.

Base Sequence ; Cell Line, Tumor ; DNA Mutational Analysis ; Enzyme-Linked Immunosorbent Assay ; Exons ; genetics ; HL-60 Cells ; Hematologic Neoplasms ; genetics ; metabolism ; pathology ; Humans ; Jurkat Cells ; K562 Cells ; Mutation ; Polymerase Chain Reaction ; Telomerase ; metabolism ; Telomere-Binding Proteins ; genetics ; metabolism ; U937 Cells

OBJECTIVETo detect the expression levels of telomere binding factor 2 (TRF2) on leukemia cell lines and primary leukemia cells.

METHODSThe expression of TRF2 mRNA was detected with quantitative real-time RT-PCR in leukemia cell lines and primary leukemia cells. The Western blot analysis was used for the detection of TRF2 protein expression.

RESULTTRF2 was overexpressed in T-cell leukemia cell lines but not in myelogenous leukemia cell lines. Significant higher expression levels of TRF2 were observed in primary leukemia cells from patients with M0 and M1 subtypes of acute myelogenous leukemia (AML) compared with normal control and other subtypes of AML.

CONCLUSIONIncreased TRF2 expression levels are found in T-cell leukemia cell lines and AML patients with poor prognosis, which suggests that TRF2 expression might be related to the prognosis of leukemia.

Adolescent ; Adult ; Female ; HL-60 Cells ; Humans ; Jurkat Cells ; K562 Cells ; Leukemia, Myeloid, Acute ; metabolism ; Leukemia, T-Cell ; metabolism ; pathology ; Male ; Middle Aged ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Telomeric Repeat Binding Protein 2 ; metabolism ; Young Adult

Beta-catenin plays a central role in Wnt signaling pathway. The aberrant localization of beta-catenin in nucleus causes the transcription of down-stream target genes, which is the pathogenesy of some solid tumours. As the expression of adheren junction on hemopoietic cells is very low, there are a few studies on beta-catenin expression in leukaemia. This study was aimed to investigate beta-catenin localization and beta-catenin mRNA expression levels in 4 leukemia cell lines so as to explore a new oncogenic mechanism and to find out a new therapeutic target. The beta-catenin localization in leukemia cell lines was detected by immunocytochemistry, the beta-catenin mRNA expression level was assayed by real-time quantitative RT-PCR. The results showed that there was aberrant localization of beta-catenin in Jurkat and Thp-1, and beta-catenin mRNA expression level was not increased in these two cell lines, however, the mRNA expression levels of Jurkat and Thp-1 were lower than those of Daudi and K562. The beta-catenin mRNA expression level was not correlated with beta-catenin aberrant localization in these 4 cell lines. It is concluded that the aberrant localization of beta-catenin may play a role in the development of some leukemia, and the mechanism resulting in beta-catenin aberrant localization not take place at transcription level.
Burkitt Lymphoma ; metabolism ; pathology ; Cell Line, Tumor ; Hematologic Neoplasms ; pathology ; Humans ; Jurkat Cells ; K562 Cells ; Leukemia, Monocytic, Acute ; pathology ; RNA, Messenger ; biosynthesis ; genetics ; Signal Transduction ; Wnt Proteins ; metabolism ; beta Catenin ; analysis ; biosynthesis ; genetics

OBJECTIVETo investigate the cytotoxic effects of mTOR inhibitor rapamycin (Rapa) and idarubicin (IDA) on human T-cell acute lymphoblastic leukemia Jurkat cell line.

METHODSThe proliferation of Jurkat cells was observed by CCK-8 assay. The combined index was analyzed by Isobologram method. Apoptosis was detected by electron microscopy and flow cytometry with Annexin V/PI staining. Protein expressions of Caspase 3, PARP, Caspase 8, Caspase 9, Akt, p-Akt, P85S6K, p-P85S6K, P70S6K, p-P70S6K, ERK1/2 and p-ERK1/2 were determined by Western blotting.

RESULTSThe IC(50) of IDA for Jurkat cells was significantly reduced when combined with 10 nmol/L rapamycin. The combined index was <1. Both electron microscopy and Annexin V/PI staining flow cytometry revealed that rapamycin significantly increased apoptotic sensitivity to IDA. The combination of IDA with rapamycin enhanced the expressions of Caspase 3, PARP, Caspase 8 and Caspase 9. Rapamycin significantly inhibited mTOR signaling upstream Akt and downstream S6K activation triggered by IDA, and the combination of the two agents led to synergistic inhibition of ERK phosphorylation.

CONCLUSIONCombination of IDA with rapamycin exerted a synergistic anti-proliferative effect and promoted apoptosis by both extrinsic and intrinsic apoptotic pathways in Jurkat cells. Inhibition of ERK phosphorylation and mTOR signaling by rapamycin may play a certain role in this synergistic effect.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Proliferation ; drug effects ; Drug Synergism ; Humans ; Idarubicin ; pharmacology ; Jurkat Cells ; metabolism ; pathology ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; metabolism ; pathology ; Proto-Oncogene Proteins c-akt ; metabolism ; Sirolimus ; pharmacology

OBJECTIVETo investigate the expression variation and significance of Skp2 and p27(kip1) during the proliferation of lymphoma cell line Jurkat cells.

METHODSThe binding of p27(kip1) and Skp2 in Jurkat cells were detected by immunoprecipitation. Jurkat cells were treated with serum starvation and release synchronization. The expression variation and subcellular localization of p27(kip1) and Skp2 were detected by subcellular fractionation, Western blot and double immunofluorescence labelling.

RESULTSThe results of immunoprecipitation suggested that p27(kip1) and Skp2 could bind each other in Jurkat cells. During the proliferation of Jurkat cells, the protein expression of p27(kip1) decreased and intranuclear p27(kip1) decreased significantly, while the Skp2 protein increased and cytoplasmic Skp2 increased significantly.

CONCLUSIONDuring the proliferation of Jurkat cells, the increased cytoplasmic synthesis of Skp2 may speed up p27(kip1) degradation via the ubiquitin-proteasome pathway, then intranuclear p27(kip1) decreases significantly, leading to an increased cell cycling activity.

Cell Nucleus ; metabolism ; Cell Proliferation ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Cytoplasm ; metabolism ; Humans ; Jurkat Cells ; Lymphoma, B-Cell ; metabolism ; pathology ; Protein Binding ; S-Phase Kinase-Associated Proteins ; metabolism

To investigate the pattern of human ERMAP gene expression in different cell lines, 15 cell lines derived from hematopoietic tumor, somatic tumor and normal tissue were chosen and were cultured; cells were harvested after culture for 12, 24, 36, 48, 60 and 72 hours; the expression of the human ERMAP was detected by using fluorescent quantitative PCR. The results showed that human ERMAP gene expression was positive in K562 cell line at interval of 12 and 24 hours, and the expression levels were (5.092 +/- 2.331) x 10(6) cps/microl RNA, (5.328 +/- 3.916) x 10(6) cps/microl RNA, respectively; ERMAP gene expression was also positive in ECV304 cell line at interval of 24 hours, and the expression level was (0.84 +/- 0.12) x 10(6) cps/microl RNA; and its expression was negative in other 13 cell lines. It is concluded that human ERMAP gene expression in ECV304 cell line was found first, and its expression in K562 cell line was further confirmed.
Blood Group Antigens ; genetics ; Butyrophilins ; Cell Line ; Cell Line, Tumor ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; HL-60 Cells ; Hematologic Neoplasms ; genetics ; pathology ; Humans ; Jurkat Cells ; K562 Cells ; RNA, Messenger ; genetics ; metabolism