Establishment of patient-like SCID mouse model by orthotopically implanting primary cultured cells from surgically-resected lung cancer tissues.

通过原位植入手术切除的肺癌组织的原代培养细胞建立类患者 SCID 小鼠模型。

• 批准号: 14571269
• 负责人: MONDEN Yasumasa
• 金额: $ 1.92万
• 依托单位: THE University of Tokushima
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571269/
• 关键词: lung cancer; primary cultured cancer cell; orthotopic implantation model; metastatic pattern; DNA microarray; RNA expression; operation sample; 同所性移植; 転移様; 同所移植モデル; 肺癌初代培養; 抗癌剤感受性試験

Background : Lymphogenous and hematogenous metastasis occurs frequently in patients with lung cancer. Suppression of these metastases provide an improvement in survival time in lung cancer patients. However, there is no patient-like animal model of these metastases of human lung cancer. We established a patient-like SCIIZ) mice model by orthotopic implantation using human non-small cell lung cancer (NSCLC) cell lines and first cultured lung cancer cells from resected tumor samples. Material and methods : Surgically-resected tissues of 5 patients with non-small cell lung cancer (NSCLC) were applied to a primary culture method using a collagen gel coated flask. We could maintain primary culture cells from 2 (FM205 and FT821cells) of 5 lung cancers, and made orthotopically implanted SOID mouse models. The size of both tumors in implanted sites of the lung increased with the time. Suspensions of 2.0x10^4 cancer cells with 10mg/ml Matrigel were injected into the left lung of SOID mice using … More a 30-gauge needle. We observed the progressive stages of development of lymphogenous and hematogenous metastasis from 2 week to 16 week in 4 lung cancer cell lines (Ma2, Ma10, Ma44-3, A549) and 2 primary culture cells(FM205 and FT821). And we evaluate a differences of RINA expression of 57 genes (anticancer drug-metabolism, -resistant enzyme, and DNA repair gene protein, etc) between implanted lung cancer tissues and lung cancer cell lines using DNA array. Results: All cell lines made tumor in the implanted site. The tumor in the implanted site of the lung became larger according to the time. Three cell lines (Ma44-3, FT821, and FM205) had the mediastinal lymph node metastasis. Ma10 cell line had no metastasis. A549 and Ma25 cell line bad both of mediastinal lymph node metastasis and intrapulmonary metastases. Ma2 cell line had solitary distant metastasis (adrenal gland, ovary, or lung). The degree of RNA expression of 57 genes in each implanted tumor correlated to that in the each cancer cell line (Pearson's correlation coefficient : r=0.8883-0.9533). Conclusion: The metastatic pattern of this model is very similar to that of clinical lung cancer, and the degree of expression of anticancer drug-related enzymes in the implanted tumor correlated to that of each cell line. We suppose that this model is useful for evaluating the inhibitory effect of anticancer drugs and the mechanism of lymphogenous and hematogenous metastasis. Less

背景：肺癌患者常发生淋巴性和血行性转移。抑制这些转移可改善肺癌患者的生存时间。然而，目前还没有类似于患者的人类肺癌转移的动物模型。我们利用人类非小细胞肺癌（NSCLC）细胞系和肿瘤切除标本中首次培养的肺癌细胞，原位植入建立了患者样sciz小鼠模型。材料与方法：5例非小细胞肺癌（NSCLC）患者手术切除组织，采用胶原凝胶包被烧瓶进行原代培养。我们可以维持5种肺癌的2个原代培养细胞（FM205和ft821细胞），并制作原位植入的solid小鼠模型。两种肿瘤在肺植入部位的大小均随时间增加而增大。将含有10mg/ml Matrigel的2.0 × 10^4个癌细胞悬浮液用30号针注射到SOID小鼠左肺。我们观察了4种肺癌细胞系（Ma2, Ma10, Ma44-3, A549）和2种原代培养细胞（FM205和FT821）在2周至16周内淋巴和血液转移的进展阶段。采用DNA阵列技术，比较了移植肺癌组织和肺癌细胞系间57个基因（抗癌药物代谢、耐药酶、DNA修复基因蛋白等）的RINA表达差异。结果：所有细胞系均在植入部位形成肿瘤。肺植入部位肿瘤随时间增大。3株细胞系（Ma44-3、FT821和FM205）发生纵隔淋巴结转移。Ma10细胞系无转移。A549和Ma25细胞系均可发生纵隔淋巴结转移和肺内转移。Ma2细胞系有孤立的远处转移（肾上腺、卵巢或肺）。每个植入肿瘤中57个基因的RNA表达度与每个癌细胞系的RNA表达度相关（Pearson相关系数：r=0.8883 ~ 0.9533）。结论：该模型的转移模式与临床肺癌非常相似，移植肿瘤中抗癌药物相关酶的表达程度与各细胞系的表达程度相关。我们认为该模型可用于评价抗癌药物的抑制作用以及淋巴和血液转移的机制。少

项目成果

Hisashi Ishikura, Kazuya Kondo, Takanori Miyoshi, Hidetaka Kinoshita, Yuji Takahashi, Haruhiko Fujino, Yasumasa Monden: "Suppression of mediastinal metastasis by uracil-tegafur or cis-diamminedichloroplatinum (II) using a lymphogenous metastatic model in

Hisashi Ishikura、Kazuya Kondo、Takanori Miyoshi、Hidetaka Kinoshita、Yuji Takahashi、Haruhiko Fujino、Yasumasa Monden：“使用淋巴转移模型通过尿嘧啶替加氟或顺式二氯二氨铂 (II) 抑制纵隔转移

Haruhiko Fujino他: "Establishment of patient-like SCID mouse model by orthotopically implanting primary cultured cells from surgically-resected lung"Oncology Reports. 10. 1709-1715 (2003)

Haruhiko Fujino 等人：“通过原位植入来自手术切除的肺的原代培养细胞建立患者样 SCID 小鼠模型”肿瘤学报告 10. 1709-1715 (2003)。

Hisashi Ishikura, Kazuya Kondo, Takanori Miyoshi, Hidetaka Kinoshita, Toshiyuki Hirose, Yasumasa Monden: "Artificial lymphogenous metastatic model using orthotopic implantation of human cancer."Ann.Thorac.Surg.. 69. 1691-1695 (2000)

Hisashi Ishikura、Kazuya Kondo、Takanori Miyoshi、Hidetaka Kinoshita、Toshiyuki Hirose、Yasumasa Monden：“使用人类癌症原位植入的人工淋巴转移模型。”Ann.Thorac.Surg.. 69. 1691-1695 (2000)

Takanori Miyoshi, Kazuya Kondo, Hisashi Ishikura, Hidetaka Kinoshita, Yasumichi Matsumori, yasumasa Monden: "SCID mouse lymphogenous metastatic model of human lung cancer using orthotopic inoculation of cancer cells."Anticancer Res.. 20. 161-164 (2000)

Takanori Miyoshi、Kazuya Kondo、Hisashi Ishikura、Hidetaka Kinoshita、Yasumichi Matsumori、yasumasa Monden：“使用癌细胞原位接种的人肺癌 SCID 小鼠淋巴转移模型。”Anticancer Res.. 20. 161-164 (2000)