Masami Niiya

Induction of IL-8 and monoclyte chemoattractant protein-1 by doxorubicin in human small cell lung carcinoma cells

We previously demonstrated doxorubicin‐induced urokinase expression in human H69 SCLC cells by the microarray technique using Human Cancer CHIP version 2 (Takara Shuzo, Kyoto, Japan), in which 425 human cancer‐related genes were spotted on glass plates (Kiguchi et al., Int J Cancer 2001;93:792–7). Microarray analysis also revealed significant induction of IL‐8, a member of the CXC chemokines. We have, therefore, extended the observation by testing the effects of doxorubicin on expression of the chemokine family and provide here definitive evidence that doxorubicin induces IL‐8 and MCP‐1, one of the CC chemokines, at least in 2 human SCLC cells, H69 and SBC‐1. IL‐8 antigen levels, measured by ELISA, were markedly increased in both H69 and SBC‐1 conditioned media after doxorubicin treatment, in parallel with mRNA levels; and this was dependent on the dose of doxorubicin. The ribonuclease protection assay, using a multiprobe template set for human chemokines, revealed induction of not only IL‐8 but also MCP‐1 in doxorubicin‐treated H69 cells. MCP‐1 antigen levels increased approximately 100‐fold in doxorubicin‐treated H69 cells. RT‐PCR using specific primers for MCP‐1 suggested that doxorubicin also induced MCP‐1 expression in SBC‐1 and SBC‐3 SCLC cells. Futhermore, CAT analysis using IL‐8 promoter implicated the PEA3 transcriptional factor, whose binding site was located immediately upstream of the AP‐1 and NF‐κB binding sites. Thus, it is suggested that doxorubicin induces IL‐8 and MCP‐1 chemokines in human SCLC cells by activating gene expression, in which at least PEA3 is involved. IL‐8 and MCP‐1 are major chemoattractants for neutrophils and monocytes/macrophages, respectively; therefore, extensive induction of IL‐8 and MCP‐1 may provoke the interaction between inflammatory/immune cells and tumor cells under doxorubicin stimulation and influence many aspects of tumor cell biology.

Incidental Detection of Acute Lymphoblastic Leukemia on [18F]Fluorodeoxyglucose Positron Emission Tomography

A 32-year-old woman was referred to our hospital because of diffuse bone pain and a high fever lasting 2 weeks. She had a normal physical examination with no sign of hepatosplenomegaly or adenopathy. The hematological examination revealed no abnormalities; the WBC count was 7,410.0/mm (neutrophils, 84.9%; eosinophils, 0.8%; basophils, 0.4%; monocytes, 1.8%; lymphocytes, 12.2%), hemoglobin was 12.1 g/dL, and platelet count was 221,000/mm. Laboratory tests showed an increased lactate dehydrogenase (409 IU/L; reference range, 120 to 240 IU/L) and C-reactive protein (22.10 mg/dL; reference range, 0.00 to 0.30 mg/dL). Computed tomography (CT) and magnetic resonance imaging found no evidence of abnormalities, while positron emission tomography (PET)/CT with [F]fluorodeoxyglucose (FDG) showed diffuse increased bone marrow uptake (Fig 1). Based on the PET/CT result, we examined a bone marrow aspirate, which revealed 98% lymphoblasts (Fig 2). The leukemic cells in the bone marrow expressed CD19, CD10, CD34, and CD13 antigens. A multiplex real-time quantitative polymerase chain reaction assay showed that the copy number of minor BCR/ABL transcripts was elevated at 1.7 10 copies. She was diagnosed with acute lymphoblastic leukemia with the Philadelphia chromosome, was treated immediately with chemotherapy including imatinib, and achieved molecular complete remission. Routinely, PET/CT is used to find a wide variety of tumors with detection sensitivities and specificities usually exceeding 90%, particularly in cancers of the lung, colon, head and neck, and melanoma. In hematological malignancies, its value for diagnosis and response assessment has been confirmed in lymphoma, myeloid sarcoma, and multiple myeloma. Previous report assessed the response of acute myeloblastic leukemia using PET/CT with [F]fluorodeoxythymidine, although the diagnostic value of PET/CT has not been evaluated thoroughly in leukemia. In several published case reports, PET detected focal bone localization of leukemia, while bone marrow aspiration detected no abnormalities. In our case, leukemic cells were localized in the bone marrow only, while hematological examination of the peripheral blood revealed no abnormalities. PET/CT might be useful for detecting leukemia in cases that are difficult to diagnose, allowing treatment initiation without delay.

Involvement of ERK1/2 and p38 MAP Kinase in Doxorubicin-Induced uPA Expression in Human RC-K8 Lymphoma and NCI-H69 Small Cell Lung Carcinoma Cells

We previously demonstrated the doxorubicin-induced urokinase-type plasminogen activator (uPA) expression in human RC-K8 lymphoma cells and NCI-H69 small cell lung carcinoma cells in which reactive oxygen species might be involved. Western blotting analysis revealed phosphorylation/activation of mitogen-activated protein (MAP) kinases, such as extracellular signal-regulated kinase (ERK) 1/2, p38 MAP kinase and stress-activated protein kinase/c-jun N-terminal protein kinase (SAPK/JNK) in doxorubicin-treated RC-K8 and H69 cells, and, therefore, we attempted to identify the MAP kinases implicated in doxorubicin-induced uPA expression by the use of their specific inhibitors. U0126, SB202190 and JNKI-1, inhibitors for MAPK kinase, (MEK) 1/2, p38 MAP kinase and SAPK/JNK, respectively, specifically and clearly inhibited their corresponding kinases. U0126 and SB202190, but not JNKI-1, almost completely inhibited the doxorubicin-induced uPA expression in both RC-K8 and H69 cells. However, U0126 rather enhanced the doxorubicin-induced activation of caspase-3 and poly ADP-ribose polymerase (PARP), and U0126 itself activated caspase-3 and PARP. Interestingly, JNKI-1 inhibited the doxorubicin-induced activation of caspase-3 and PARP. Therefore, doxorubicin treatment activates the above three kinases, but different MAP kinase signaling is responsible in the doxorubicin-induced caspase activation and expression of uPA. Thus, we could possibly manipulate the direction of doxorubicin-induced MAP kinase activation and the effects of doxorubicin on the tumor cell biology by the use of MAP kinase inhibitors.

Prediction of number of apheresis procedures necessary in healthy donors to attain minimally required peripheral blood CD34+ cells

BACKGROUND: Allogeneic peripheral blood stem cell (PBSC) transplantation is widely performed as a curative therapy for hematopoietic malignancies. Donors for PBSC harvest (PBSCH) are usually healthy subjects and undergo granulocyte–colony‐stimulating factor treatment and apheresis procedures. A considerable proportion of donors experience poor mobilization, necessitating additional harvesting or marrow collection or remobilization. Although some characteristics have been reported to correlate with poor mobilization, they may not be taken into account in selecting PBSC donors. To protect healthy donors, it is preferable to predict the number of apheresis procedures needed for PBSCH before the procedure is initiated.

Induction of urokinase-type plasminogen activator, interleukin-8 and early growth response-1 by STI571 through activating mitogen activated protein kinase in human small cell lung cancer cells

We previously demonstrated the simultaneous induction of urokinase-type plasminogen activator and interleukin-8, a CXC chemokine, in doxorubicin-treated human NCI-H69 small cell lung cancer cells in which extracellular signal-regulated kinase 1/2 and p38 mitogen-activated protein kinase might be involved. NCI-H69 cells expressed one of the receptor tyrosine kinases, c-Kit, and STI571 inhibited the cell growth and stem cell factor-induced phosphorylation of c-Kit. We therefore investigated the effects of STI571 on the expression of urokinase-type plasminogen activator and interleukin-8 in NCI-H69 cells. Microarray analysis revealed the gene induction of not only urokinase-type plasminogen activator and interleukin-8, but also early growth response-1 in STI571-treated cells. Treatment with STI571 resulted in the induction of phosphorylation of all three mitogen-activated protein kinases, such as extracellular signal-regulated kinase 1/2, p38 mitogen-activated protein kinase and stress-activated protein kinase/c-jun N-terminal protein kinase. U0126, an inhibitor against extracellular signal-regulated kinase 1/2, however, only inhibited the STI571-induced interleukin-8 accumulation. Urokinase-type plasminogen activator and interleukin-8 are important biological factors in tumor cell regulation; STI571 may therefore influence many aspects of tumor cell biology through inducing urokinase-type plasminogen activator and interleukin-8, in which the induction of early growth response-1 expression and extracellular signal-regulated kinase 1/2 phosphorylation might be involved.