特異的DNA増幅反応法を用いたヌードマウスにおけるヒト転写腫瘍細胞の定量的検出

取得学位 : 博士（医学）, 学位授与番号 : 医博甲第1022号,学位授与年月日：平成4年3月25日,学位授与年：199

胸膜播種に対する光力学治療と抗VEGF中和抗体を併用した治療効果の検討

癌性胸膜炎モデルを設定し抗VEGF中和抗体を用いた治療実験を試みた。実験動物は8週齢雌のヌードラットを用い、移植腫瘍はヒト低分化型肺腺癌細胞株PC-14を使用した。レスピレータ管理下に左側開胸を行ない、第1モデルでは肺摘除後、壁側胸膜下にPC-14を1x10^7個移植した。第2モデルでは、開胸時胸腔内にPC-14を直接散布した。移植後1週目にラットを無作為に治療群・非治療群の2群に分け、治療群に対し抗VEGF中和抗体を250μg/bodyにて週2回のべ1mgを第1モデルでは胸腔内、第2モデルでは腹腔内投与し、腫瘍移植後4週目にautopsyした。第1モデル系では、全ラットに腫瘍の生着および多発性結節性種瘤の形成が確認された。胸膜への転移形成は組織学的にも確認された。第1モデルでは、胸膜下移植腫瘍のサイズは、治療群で縮小傾向を示した。播種結節数は治療群で有意に少なく、播種のひろがりも中和抗体により抑制された。胸膜下移植腫瘍内の血管密度は中和抗体投与群で低下し、腫瘍細胞での自己分泌型運動因子受容体(AMFR)の発現性が中和抗体投与群において減弱していた。Western blot解析でも同様の結果が得られ、胸膜播種の形成にVEGFとAMFRが協調的に関与している可能性が示された。第2モデル系では、非治療群8匹中5匹に胸水が認められ、中2匹が悪性胸水であったのに対し、中和抗体投与群に胸水貯留例は認めなかった。以上、ヌードラットを用いた2種類の癌性胸膜炎モデルの検討から、VEGFの癌性胸膜炎への関与が支持され、その機序として血管膜透過性以外に、胸膜における脈管を介した転移経路の関与が示された。VEGFは本病態に関わる重要なKey factorであり、播種および胸水の防止に有効性の期待できる分子標的と判断された。なお、フォトフリンを用いた光力学療法との併用による治療実験を試みたが、治療群におけるラットは全例死亡し、実験を中断しその原因を検討している。We assessed the inhibitory effect of anti-vascular eridothelial growth factor (VEGF) neutralizing antibody on the formation of pleural dissemination and pleural effusion using two types of in-vivo model systems. Immune-deficient rats were inoculated with PC-14 cells into (1) a subpieurai space of the parietal pleura after pneumonectomy or (2) into the thoracic cavity directly The rats bearing tumor cells were randomly separated into two groups : non-treatment and treatment with anti-VEGF neutralizing antibody groups. In the treatment group, the antibody was administered at a dose of 250 /μg twice weekly (total 1 mg) from the 7th day after the tiimor inoculation! At the time of the autopsy (4 weeks after the tumor inoculation), all rats developed gross pleural dissemination with/without malignant effusions. In the first model system, despite no significant difference in the mean volume of the primary subpleurai tumors between the groups, the degree of dissemination was suppressed in the treatment group. The immunohistochemical examination showed less tumor vasculature and less frequency of the autocrine motility factor receptor expression in cancer cells at the primary site of the treatment group. In the second model, the inhibitory effect on pleural dissemination was not clear. These results demonstrated the possible association of VEGF with the development of pleural dissemination/metastasis in the context of blood/lymphatic routes and cancer cell motility.研究課題/領域番号:12671302, 研究期間(年度):2000-200

Results of initial operations in non-small cell lung cancer patients with single-level N2 disease

金沢大学医学部附属病院呼吸器外科Background. There is still debate regarding the use of surgery in the management of non-small cell lung cancer patients with N2 disease. The primary aim of the present study was analysis of the results of initial operations in non-small cell lung cancer patients with single-level N2 disease. Methods. Ninety-four patients with the disease who underwent initial surgery consisting of complete resection of the primary site plus systematic lymphadenectomy were examined. We also immunohistochemically examined lymphatic vessel density and vascular endothelial growth factor-C expression. Results. The overall 5- and 10-year survival rates for the 94 patients were 27.1% and 12.1%, respectively, with a median survival of 22 months. When stratified by skipping status, the 5-year survival rates for the patients in skip-N2 and non-skip-N2 groups were 33.4% and 19.8%, respectively (p = 0.0189). Skip metastasis, T factor, subcarinal lymph node metastasis, and adjuvant chemotherapy were recognized as independent prognostic indicators. In both skip-N2 and non-skip-N2 groups, distant relapse was the dominant pattern of recurrence. Although the peritumoral lymphatic vessel density was associated with vascular endothelial growth factor-C expression in tumors, the lymphangiogenic profile appeared to be different between skip-N2 and non-skip-N2 tumors, suggesting different nodal metastatic process. Conclusions. Lung cancer patients with single-level N2 disease are an oncologically heterogeneous cohort. Although further studies involving randomized comparisons are required, the poor outcomes found here indicate that the initial operation has yet to be validated for patients with this disease. © 2006 by The Society of Thoracic Surgeons

Evaluation of atherosclerotic lesions using dextran- and mannan–dextran-coated USPIO: MRI analysis and pathological findings

Magnetic resonance imaging (MRI) can detect atherosclerotic lesions containing accumulations of ultrasmall superparamagnetic iron oxides (USPIO). Positing that improved USPIO with a higher affinity for atherosclerotic plaques would yield better plaque images, we performed MRI and histologic studies to compare the uptake of dextran- and mannan–dextran-coated USPIO (D-USPIO and DM-USPIO, respectively) by the atherosclerotic walls of rabbits. We intravenously injected atherosclerotic rabbits with DM-USPIO (n = 5) or D-USPIO (n = 5). Two rabbits were the controls. The doses delivered were 0.08 (dose 1) (n = 1), 0.4 (dose 2) (n = 1), or 0.8 (dose 3) (n = 3) mmol iron/Kg. The dose 3 rabbits underwent in vivo contrast-enhanced magnetic resonance angiography (MRA) before and 5 days after USPIO administration. Afterwards, all animals were euthanized, the aortae were removed and subjected to in vitro MRI study. The signal-to-noise ratio (SNR) of the aortic wall in the same region of interest (ROI) was calculated in both in vivo and in vitro studies. Histological assessment through measurement of iron-positive regions in Prussian blue-stained specimens showed that iron-positive regions were significantly larger in rabbits injected with DM- rather than D-USPIO (P < 0.05) for all doses. In vivo MRA showed that the SNR-reducing effect of DM- was greater than that of D-USPIO (P < 0.05). With in vitro MRI scans, SNR was significantly lower in rabbits treated with dose 2 of DM-USPIO compared with D-USPIO treatment (P < 0.05), and it tended to be lower at dose 3 (P < 0.1). In conclusion, we suggest that DM-USPIO is superior to D-USPIO for the study of atherosclerotic lesions in rabbits

Surgical Results in T2N0M0 Nonsmall Cell Lung Cancer Patients With Large Tumors 5 cm or Greater in Diameter: What Regulates Outcome?

金沢大学医学部附属病院呼吸器外科Background: We assessed the surgical results along with the clinical and biological features of nonsmall-cell lung cancer (NSCLC) patients with localized large tumors. Methods: The study population consisted of 86 NSCLC patients who underwent complete resection of tumors 5 cm or larger in diameter in stage IB (T2N0M0). We immunohistochemically assessed the expression of angiostatin and endostatin. Results: The median tumor size was 6.0 cm (range, 5 to 14 cm). The operative procedures used were lobectomy in 71 cases, bilobectomy in 8 cases, and pneumonectomy in 11 cases. Fifty patients (58.1%) relapsed during the mean follow-up period of 33.6 ± 4.5 months. The median disease-free interval was 9 months. Of 44 recurrent patients whose disease-free interval could be identified, 25 patients (56.8%) relapsed within 12 months after the operation. The overall 5- and 10-year survival rates were 42.0% and 24.2%, respectively. Multivariate analysis showed that the degree of pleural involvement and angiostatin expression within the tumor were independent prognostic indicators. The endostatin expression within tumors also had a weaker relationship with outcome. Conclusions: Long-term surgical results were poor and early relapse was common in this cohort. In addition to pleural involvement, the tumor-induced expression of angiostatin and endostatin merit further investigation to gain possible insights into selection of patients who will benefit from surgery as the first line treatment. © 2006 The Society of Thoracic Surgeons