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鹿児島大学

Object: To investigate the usefulness of ultraviolet-microscopic spectroscopy （UV-MS） of unstained cells by liquid-based cytology （LBC） to objectively differentiate non-cancer from cancer cells. Study Design: Cultured cells were used as the sample cells: 100 non-cancer cells and 200 cancer cells. The sample measurement region was a 166.4-μm2 area in the nuclear region of sample cells. On UV-MS, data of 260, 280, 300, 320, and 340 nm were extracted from the transmittance spectrum of 260-350-nm ultraviolet （UV） wavelengths and analyzed. Result: At a 300-nm UV wavelength, transmittances in non-cancer and cancer cells were 79.7±5.0 and 64.1±5.0%, respectively, being signifi cantly lower in cancer cells （P, 弘前医学. 65, 2014, p.82-94

Claudin is one of tight junction proteins which connect with the actin cytoskeleton and participate in the intracellular signaling. However, the significance of claudin in pancreatic cancer is understood not yet extensively. We examined the relationship between claudin-1 and invasion in PANC-1 and MIA PaCa-2 human pancreatic cancer cells, and investigated the functions of claudin-1 in invasive growth of pancreatic cancer cells. Claudin-1 knockdown by siRNA （claudin siRNA） affected the subcellular localization in the pancreatic cancer cells, and claudin-1 siRNA increased numbers of invasive pancreatic cancer PANC-1 and MIA PaCa-2 cells. Claudin-1 siRNA did not significantly affect expression levels of β-catenin, E-cadherin, α-smooth muscle actin, Bcl-2, and Bax in PANC-1 and MIA PaCa-2 cells. In addition, claudin-1 siRNA showed no significant change in the cell proliferation. We concluded that claudin-1 is significantly associated with invasive growth of human pancreatic cancer cells., 弘前医学. 63, 2012, p.127-135

学童期にがんについて学ぶことは、将来にわたって予防検診といった知識が役に立つと同時に、彼らの身近な親族ががんと診断されることが珍しくはないという現状から効果的であるといえる。そこで著者らは小学校高学年の児童を対象とした教材を開発し、その効果の検証を行った。本稿では、開発の過程と効果の検証の結果を報告するとともに、そこから導かれるがん教育における政策的な提言を行うことを目的とした。

Previous studies have assessed the efficacy and safety of combined treatment with irinotecan (7-ethyl-10-[4-[1-piperidino]-1-piperidino]carbonyloxycamptothecin, CPT-11) and S-1, containing tegafur, a prodrug of 5-fluorouracil, in the treatment of colorectal and gastric cancer. The objective of this study was to describe the interaction between CPT-11 and S-1 in 4 patients with colorectal cancer. Coadministration of S-1 changed the pharmacokinetic behavior of CPT-11 and its metabolites. In particular, maximum plasma concentration (Cmax) and area under the plasma concentration curve (AUC) of 7-ethyl-10-hydroxycampothecin (SN-38) was markedly decreased by coadministration of S-1. For SN-38, the median ratio of Cmax and AUC with S-1 to those without S-1 was median 0.34 (range 0.24-0.78) and 0.56 (range 0.23-0.68), respectively. A markedly difference in drug interaction among individual patients was observed. We conclude that the plasma concentration of SN-38 was decreased by oral administration of S-1 in patients with colorectal cancer. This observation might be important for clinical decisions regarding combination therapy.

Purpose The purpose of this study was to examine the regulation of renal organic ion transporters in cisplatin-induced acute kidney injury (AKI) and its relation with indoxyl sulfate (IS), a uremic toxin. Methods The IS concentrations in the serum and kidney were monitored by high-performance liquid chromatography. Uptake of p-aminohippuric acid, estrone-3-sulfate and tetraethylammonium were examined using renal slices. Real-time PCR and immunoblotting were performed to examine the mRNA and protein expression of rOATs, rOCTs and rMATE1 in the kidney, respectively. Results The serum and renal IS levels were markedly elevated in cisplatin-treated rats. However, this effect was largely reversed by administration of AST-120, an oral charcoal adsorbent. The functions of renal basolateral organic anion and cation transporters were reduced in cisplatin-treated rats. The levels of mRNA and protein corresponding to rOAT1, rOAT3, rOCT2 and rMATE1, but not rOCT1, were depressed in the kidney of cisplatin-treated rats. Administration of AST-120 to cisplatin-treated rats partially restored the function and expression level of these transporters. Conclusions Cisplatin-induced AKI causes down-regulation of renal organic ion transporters accompanied by accumulation of serum and renal IS. IS could be involved in the mechanism of down-regulation of rOAT1, rOAT3 and rMATE1 under cisplatin-induced AKI.

Objective: The purpose of this study was to prospectively evaluate the efficacy and safety of micafungin (MCFG) in empirical therapy for febrile neutropenic patients for whom antibiotic therapy was not effective for hematological malignancies. Patients and Methods: Twenty-three hematological patients aged 27-82 years with febrile neutropenia for whom antibiotic therapy was not effective were enrolled in this study and responses to treatment were evaluated. Results: Treatment success rate was 73.9%. Treatment success rates by primary diagnosis were 77.8% in patients with AML, 50.0% in patients with NHL and 87.5% in patients with other diseases. Moreover, MCFG at a dose of 100 mg or more have a tendency to be effective. One or more adverse events occurred in five (27.7%) of the patients during the study. All of these adverse events were below grade 2 toxicity. Conclusions: Although the number of patients studied was limited, MCFG as a monotherapy seems to be effective and safe as an empirical therapy in patients with febrile neutropenia. However, further investigation using large-scale studies is needed. This study demonstrated the clinical efficacy and safety of MCFG in patients with febrile neutropenia and with hematological malignancies.

We have previously described a method for adoptive immunotherapy of cancer based on antigen-specific T(h)1 cells. However, efficient induction of anti-tumor responses using T(h)1 cells remains a formidable challenge, especially for MHC class II-negative tumors. In the present study, we sought to develop a novel strategy to eradicate established tumors of the MHC class II-negative, ovalbumin (OVA)-expressing EG-7 cells. Tumor-bearing mice were intradermally treated with OVA-specific T(h)1 cells, combined with the model tumor antigen (OVA), near the tumor-draining lymph node (DLN). We found that tumor growth was significantly inhibited by this strategy and approximately 50-60% of tumor-bearing mice were completely cured. Tumor eradication was crucially dependent on the generation of OVA/H-2K(b)-specific CTLs in the tumor DLNs and tumor site. The injected T(h)1 cells were mainly distributed in tumor DLNs, where they vigorously proliferated and enhanced the activation of dendritic cells. Strikingly, we also found that the accumulation of CD4(+)CD25(+) regulatory T cells (Tregs) was significantly inhibited in tumor DLNs by T(h)1 cell adjuvant therapy and this abrogation was associated with IFNgamma secreted by T(h)1 cells. These results identify T(h)1 cell adjuvant therapy combined with tumor vaccination as a novel approach to the treatment of human cancer.

A 39-year-old Japanese woman presented with a swollen right hand and a right-sided pneumothorax. Chest CT revealed bilateral multiple pulmonary thin-walled cysts measuring

Interleukin (IL)-12 is a 70-kDa cytokine comprised of two disulfide-linked proteins (p35 and p40) and is essential for the initiation of effective immune response. Granulocyte-colony stimulating factor (G-CSF) affects the balance in the production of anti-inflammatory cytokines. We investigated the serum IL-12 p40 and IL-12 Mix (p40 and p70) production in 28 patients with B-cell lineage non-Hodgkin's lymphoma (NHL) treated with chemotherapy (e.g., CHOP regimen) with or without G-CSF administration and eight healthy volunteers. We found that serum levels of IL-12 p40 (191.2 +/- 150.0 pg/mL) and IL-12 Mix (277.4 +/- 274.5 pg/mL) in the patients before chemotherapy were higher than those in the healthy volunteers (IL-12 p40: 76.4 +/- 25.3 pg/mL, IL-12 Mix: 48.5 +/- 33.4 pg/mL) (P = 0.04 and 0.02, respectively). Next, we examined the serum IL-12 p40 and IL-12 Mix levels in nine patients receiving chemotherapy with administration of G-CSF (CG group, n = 9) and without G-CSF (C group, n = 9). Serum IL-12 p40 and IL-12 Mix levels were decreased on 10 d after chemotherapy in both groups, and those in CG groups were significantly lower than those in C group. These results indicated that administration of G-CSF decreased serum IL-12 p40 and IL-12 Mix levels. Overall survival (OS) at 24 months was not significantly different in the two groups (58.3% in group C vs. 80.0% in group CG, P = 0.67). However, the survival rate of patients at clinical stages III and IV in CG group (n = 6, 66.0%) was significantly better than that of patients in C group (n = 4, 25.0%) (P = 0.02). Long-term administration of G-CSF appears to influence the survival rate by reducing immunosuppressive IL-12 p40 production.

A novel metal-chelating system comprising a 4-dimethylamino- pyridine and two histidine appendages was synthesized. The two histidines were introduced by different manners; one through an amide linkage and other via a secondary amino linkage. ESR spectrum suggested a distorted pentacoordinate configuration of the copper complex of the ligand. The iron complex of the ligand had oxygen-activating property as shown by ESR spin trapping and DNA-cleaving activity as evaluated by experiments using pUC19 DNA.

The serine/threonine kinase, Akt (protein kinase B) plays a central role in the regulation of intracellular cell survival. Recently, we demonstrated that the proto-oncogene TCL1, overexpressed in human T-cell prolymphocytic leukemia, is an Akt kinase co-activator. Tightly restricted TCL1 gene expression in early developmental cells suggested that the TCL1 gene is regulated at a transcriptional level. To characterize how TCL1 gene expression is regulated, we cloned the 5'-promoter of the TCL1 gene located at human chromosome 14q32. The 5'-TCL1 promoter region contains a TATA box with cis-regulatory elements for Nur77/NGFI-B (nerve growth factor-responsive element (NBRE), CCAAGGTCA), NFkappaB, and fork head transcription factor. Nur77/NGFI-B, an orphan receptor superfamily transcription factor implicated in T-cell apoptosis, is a substrate for Akt. We hypothesized that TCL1 transactivity is regulated through Akt-induced phosphorylation of Nur77/NGFI-B in vivo. In an electrophoretic mobility shift assay with chromosomal immunoprecipitation assays, wild-type Nur77, but not S350A mutant Nur77, could specifically bind to TCL1-NBRE. A luciferase assay demonstrated that TCL1-NBRE is required for inhibition of TCL1 transactivity upon nerve growth factor/platelet-derived growth factor stimulation, which activates Akt and phosphorylates Nur77. Using a chromosomal immunoprecipitation assay with reverse transcription-PCR, nerve growth factor stimulation inhibited binding of endogenous Nur77 to TCL1-NBRE, in turn, suppressing TCL1 gene expression. The results together establish that TCL1-NBRE is a novel negative regulatory element of Nur77 (NGFI-B). To the best of our knowledge, TCL1-NBRE is the first direct target of Nur77 involving the regulation of intracellular cell death survival. This Akt-induced inhibitory mechanism of TCL1 should play an important role in immunological and/or neuronal development in vivo.

As most solid tumors are hypoxic, dendritic cells (DC) in solid tumors are also exposed to hypoxia. While many adaptation responses of tumor cells to hypoxia are known, it is yet to be determined how hypoxia affects the functions of DC. To explore the effects of hypoxia on the functions of DC, we compared the expression of surface markers, cytokines, chemokine receptors and matrix metalloproteinases (MMP) of human monocyte-derived DC (hmDC) differentiated under hypoxia to those differentiated under normoxia. Both groups of hmDC expressed similar levels of surface markers and cytokines. However, expression of MMP-9 and membrane type-1-MMP, as well as migrating activity, was significantly suppressed in hmDC differentiated under hypoxia compared with their normoxia counterparts. We also demonstrated that trichostatin A restored the production of MMP-9 in hmDC, under hypoxia. Collectively, our findings show that a hypoxic microenvironment suppresses the production of MMP in hmDC, most probably through the deacetylation of promoter regions of MMP, thus suppressing the migrating activity of hmDC. Our results suggest that the hypoxic microenvironment in solid tumor tissues may suppress the function of DC

Epithelial-mesenchymal transition (EMT) describes a process occurring during development and oncogenesis by which epithelial cells obtain fibroblast-like properties and show reduced cell adhesion and increased motility. In this report, we demonstrated typical EMT in human mammary epithelial MCF10A small interfering (si)RNA gelsolin-knockdown cells. EMT was characterized by fibroblastic morphology, loss of contact inhibition and focus formation in monolayer growth, enhanced motility and invasiveness in vitro, increased actin filaments, overexpression of RAC, activation of both extracellular signal-regulated kinase and AKT, inactivation of glycogen synthase kinase-3, conversion of cadherin from the E- to N-type and induction of the transcription factor Snail. These results suggested that gelsolin functions as a switch that controls E- and N-cadherin conversion via Snail, and demonstrated that its knockdown leads to EMT in human mammary epithelial cells and possibly to the development of human mammary tumors.

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Our previous studies demonstrated that introduction of a dominant negative H-ras mutant, N116Y, inhibits the growth of various types of cancer cells in vitro. In this study, we tested the efficacy of N116Y in blocking the growth of esophageal cancer cells using an adenoviral vector. Infection with N116Y adenovirus, (AdCMV-N116Y), in which N116Y expression is driven by the cytomegalovirus promoter, significantly reduced the in vitro growth of all esophageal cancer cell lines studied. Esophageal cancer cells that contained wild-type K-ras and H-ras (TE8, SGF3, SGF7) were more sensitive to AdCMV-N116Y than HEC46 cells that expressed mutant K-ras protein.Most importantly, direct injection of AdCMVN116Y into TE8-or SGF3-induced tumors in nude mice suppressed their growth significantly. To examine the suppressive mechanism of N116Y, cell cycle profile and the activation of extracellular signal-regulated kinase 2 (Erk2) were examined by flow cytometry and Western blot analysis, respectively. In TE8 cells, progression into S phase was clearly blocked after infection with AdCMV-N116Y. Infection with AdCMV-N116Y did not strongly suppress the activation of Erk2 after EGF stimulation in serum-starved HEC46 cells, whereas it completely suppressed activation in TE8, SGF3 and SGF7 cells. Our observations suggest that N116Y reduces growth of human esophageal cancer cells and suppresses the activation of Erk2; they also indicate that N116Y is a potential candidate gene for human esophageal cancer gene therapy.

Human transitional cell carcinomas of the bladder frequently reveal chromosomal abnormalities that span a range between chromosome 9p12 and 9qter, even at early stages of bladder carcinogenesis. Because the gene that encodes an actin-regulatory protein, gelsolin, is localized in chromosome 9q33, we examined the expression of gelsolin in a number of human bladder cancer cell lines and tissues. In all 6 cell lines and in 14 of the 18 tumor tissues (77.8%), gelsolin expression was undetectable or extremely low in comparison with its expression in normal bladder epithelial cells. Furthermore, upon the introduction of the exogenous human or mouse authentic gelsolin cDNA into a human bladder cancer cell line, UMUC-2, gelsolin transfectants of UMUC-2 greatly reduced the colony-forming ability and the tumorigenicity in vivo. These results suggest that gelsolin plays a key role as a tumor suppressor in human urinary bladder carcinogenesis.

弘前医学. 65, 2014, p.113-114

Intraarterial chemotherapy has been used to treat head and neck cancers for more than 30 years. Several reports confirm that the intraarterial infusion chemotherapy is feasible for squamous cell carcinomas of the oral cavity. However, most case showed good local response, there were some poor prognosis groups. Therefore, we started our new protocol from 1999, in which the drug distribution is evaluated using an IVR-CT system and the chemotherapy is combined with medium-dose conformal radiation therapy. This new protocol significantly improved the local response of the primary tumor.

ルーテル学院大学, 2007

Human HOX genes consist of 39 genes and encode transcription factors that function as master developmental regulators. We hypothesized that the misexpression of HOX genes was associated with carcinogenesis and malignant progression. The expression levels of 39 HOX genes in 31 human oral squamous cell carcinoma (SCC), 11 dysplasia, and 10 normal mucosa tissues were quantified by the real-time RT-PCR method. The expression levels of 18 HOX genes in the SCC tissues were significantly higher than those in the normal mucosa tissues. The dysplasia tissues showed higher expression of HOXA2, A3, B3, and D10 than normal mucosa tissues whereas they showed lower expression of HOXA1, B7, B9, and C8 than SCC. The SCC with lymph node metastasis showed high expression of HOXC6 compared to the SCC without it. These results suggest that misexpressions of particular HOX genes are implicated in the development of oral dysplasia and SCC.

授与大学:弘前大学; 学位種類:博士（医学）; 授与年月日:平成6年3月23日; 学位記番号:医博甲第987号

博士論文, 絨毛癌細胞に対する,より効果的な加温条件を求めるため,種カの条件下でのBeWo株の細胞増殖およびβ HCG産生の変化を検討した.1.加温条件の違いによる効果について培養4日目に39℃,41℃,43℃に加温した.43℃,1時間施温時の効果は,39℃,120時間加温時と同程度でかつ41℃,24時間加温時より大きく,加温効果が43℃にて著しく増強することが明らかとなった.2.加温時期の違いによる効果について対数増殖期の4日目,5日目,6日目に43℃,1時間または3時間加温した.増殖抑制は4日目が最大で,加温開始時期が遅れるにつれて加温効果の減少が認められた.3.MTXと併用した場合の加温効果について培養4日目からMTX10^^Mを48時間添加し,加温(43℃,1時間)開始の時期は,MTX添加開始から除去後10時間までの種々の時期に設定した.その結果,併用効果はMTX添加開始後24時間から除去後4時間以内が最大であり,またβ HCGの産生は細胞増殖抑制効果の大きい穫高値を持続し,増殖抑制効果が小さくなるにつれて漸減することが示された.以上の結果,BeWoの熱感受性は43℃で急激に増大し,他の細胞株と同様の加温効果が期待できると思われる.またBeWoは対数増殖期の各時期およびMTX添加および除去後の時間により熱感受性が異たっており,細胞増殖速度の減少およびMTXによる細胞周期の変化が熱感受性の重要た因子となることをうかがわせた.また加温のみでは,β HCGの産生先進が認められなかったのに対し,MTXと併用した場合には,β HCGの産生光進が高値で持続した.これは,MTXによる効果と加温による効果が各々異なる機序により引き起こされることを示している., In order to elucidate the effect of hyperthermia, gestational choriocarcinoma cell line (BeWo) was studied with respect to cell growth and hormone (β-HCG) production in various conditions of hyperthermia. 1. BeWo was heated in the 39.0-43.0℃ temperature range on Day 4. The cell growth was reduced significantly at 43℃ for I hour. The same effect was observed at 39℃ for 120 hours. And this effect was more severe than that at 41℃ for 24 hours. 2. BeWo was heated at 43℃ on Day 4, Day 5, and Day 6 during the logarithmic growth phase. Maximum suppression of the cell growth was observed on Day 4, and the effect was reduced with the length of time that heat application was delayed. 3. MTX (10^M) was added for 48 hours beginning on Day 4, and heat (43℃ for I hour) was applied at various points during or after MTX exposure. Heat showed an adjuvant effect with MTX when heat was applied between 24 hours after administration of MTX and 4 hours after removal of MTX. These findings strongly suggest that the heat sensitivity of BeWo is greatly related to the cell cycle. Elevated HCG production was not observed following heat alone, but observed following simultaneous heat and MTX administration. To elucidate the mechanism involved here, further studies will be needed.

『消化器扁平上皮癌の最先端多戦略治療拠点形成 : 文部科学省 : 21世紀COEプログラム』 pp141-146