Abstract. The purpose of this study was to analyze the effects of bufalin on the gene expression of K562 cells and on the expression of BMI-1 pathway constituents in K562 cell apoptosis. K562 cells were treated with bufalin, and the inhibition rate and apoptosis were detected by an MTT assay, flow cytometry and a microarray assay. BMI-1, p16 INK4a and p14 ARF were examined by quantitative polymerase chain reaction (qPCR). Bufalin induced significant changes in the gene expression of the K562 cells; 4296 genes were differentially expressed, 2185 were upregulated and 2111 were downregulated. The most upregulated genes were associated with transcription regulation, while the most downregulated genes were associated with the non-coding RNA metabolic processes and DNA repair. qPCR analysis demonstrated that BMI-1 was overexpressed in the K562 cells. Bufalin is able to downregulate BMI-1 expression levels in K562 cells prematurely and cause an increase in the expression levels of p16 INK4a and p14 ARF . Moreover, bufalin downregulated BCR/ABL expression levels in a time-dependent manner, and the expression of BCR/ABL was not associated with the upregulation or downregulation of BMI-1 expression. Bufalin may induce K562 cell apoptosis by downregulating BMI-1 expression levels and accordingly upregulating the expression levels of p16 INK4a and p14 ARF . Bufalin may also induce K562 cell apoptosis via downregulating BCR/ABL expression levels, and this pathway may be independent of the BMI-1 pathway.
T-lymphoblastic lymphoma (T-LBL) is a highly aggressive malignancy originating from T-lymphocyte precursors. Incidence is highest in children and adolescents. T-cell receptor (TCR) gene rearrangement is usually present. TCR gene rearrangement-negative cases are considered rare. Here, we investigated the clinicopathological features, differential diagnosis, therapy, and prognosis of TCR gene rearrangement-negative T-lymphoblastic lymphoma/leukemia (T-LBL/ALL) by case report and literature review. An 18-year-old male with polyglandular lymphadenopathy underwent an excisional lymph node biopsy and bone marrow aspiration that disclosed diffuse distribution of round, small to medium sized cells with scant cytoplasm, delicate chromatin, and frequent mitotic figures. Immunophenotyping showed expression of TDT, CD3, CD7, and CD5, no CD34, CD20, CD56, bcl-6, CD4, CD8, or MPO in lymph node tissue. Immunohistochemical staining for pathological consultation was performed by Streptavidin peroxidase (SP) method, EB virus coded small RNA (EBER) tested by in situ hybridization (ISH), (EBER-ISH). And flow cytometry of bone marrow aspirate showed that tumor cells expressed CD3, CD5, CD7; partial expression of CD2, CD10, CD38, TDT; and no expression of CD1a, CD34, CD4, CD8, mCD3, CD33, CD56, CD19, CD79a, HLA-DR and MPO. These findings led to the diagnosis of T-LBL/-ALL. Molecular genetic testing showed no TCR gene rearrangement. The patient received chemotherapy consisting of vinorelbine, pirarubicin, cyclophosphamide, asparaginase, and prednisone. Prophylactic chemotherapy of the central nervous system and radiotherapy of the mediastinum were also given. And responded to combined chemotherapy and radiotherapy. Although T-LBL/ALL typically features TCR gene rearrangement, rare cases without rearrangement may occur. Diagnosis is based on clinical characteristics, histopathology, immunotyping, and molecular genetics.
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