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In vivo Invasion Assay and Enhancement of Differentiation of Low Passage Cultured Cells Derived from Human Non-Small Cell lung Carcinoma Using De-epithelialized Rat Tracheas Transplanted into Nude Mice.

使用移植到裸鼠的去上皮大鼠气管进行人非小细胞肺癌低传代培养细胞的体内侵袭测定和增强分化。

基本信息

批准号：
01570777
负责人：
SHIBA Mitsutoshi
金额：
$ 1.41万
依托单位：
Chiba University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1990
项目状态：
已结题

项目摘要

The investigation of human tumors using nude mouse xenograft is capable of taking into account the host reactions, whereas this is impossible in in vitro system. Nude mice offer an invasion environment similar to that of humans; however, s.c. injection procedures for conventional xenotransplantation into nude mice have encountered problems with regard to long latency periods low tumor take rates. In this project, human lung tumor-derived cell lines with low passage generation were transplanted into nude mice to determine their growth behavior and invasive potential. Six cell lines were inoculated into deepithelialized rat tracheas (5 x 10^5 cells/trachea). After cell inoculation, the tracheas were sealed and transplanted into the subcutis of nude mice. In a parallel experiment, these cell lines (1 x 10^6 cells) were injected s.c. In the subcutis, the tumor take rate was only 13% with a long latency period of 18 weeks, and 3 cell lines did not show any invasive growth to the surrounding … More tissue. In rat tracheas, all cell lines proliferated within 3 weeks, and 4 of them showed invasive growth to the tracheal wall within 1-2 weeks. Cells growing in the tracheal wall showed higher (^3H) thymidine labeling indexes and greater atypia, such as larger nuclei and prominent nucleoli, than those in the tracheal lumen. The s.c. tumor take rate correlated with the incidence of invasive growth to the tracheal wall. The survival of the patients originally bearing the six tumors also correlated closely with the invasive potential of this system. In tracheal transplants, all cell lines showed comparable or higher degree of differentiation than in the primary lesion. While most cell lines showed lower degree of differentiation in the mouse subcutis. The surfaces of tracheal transplants repopulated with the cells derived form squamous cell carcinomas were initially lined by a layer of flattened cells 2 to 3 cells thick. While those repopulated with the cells from 3 cases of adenocarcinoma were initially lined by simple epithelia consisted of cuboidal-, round-, and columnar-shaped cells, respectively. Thereafter, those cell lines revealed pseudostratified pattern, papillary growth pattern, and cribriform pattern, respectively. Ultrastructurally, they showed structures resembling ciliated bronchial epithelial cells, non-ciliated bronchiolar epithelial cells, and bronchial goblet cells, respectively.These results indicate that the system using low passage cell lines can evaluate the invasive potential shortly after the inoculation; of a relatively small number of cells and can be used as a clinically reliable biological invasion assay. And, histological examination of cultured cells using this system was considered to be advantageous for evaluation of differentiation because it provided unique proliferation patterns unobserved in primary tumors or tumors formed after subcutaneous direct cell injections. Less

利用裸鼠异种移植物研究人类肿瘤能够考虑到宿主的反应，而这在体外系统中是不可能的。裸鼠提供了与人类相似的入侵环境；然而，常规异种移植的sc注射程序在裸鼠身上遇到了潜伏期长、取瘤率低的问题。本项目将低传代的人肺肿瘤来源细胞系移植到裸鼠体内，研究其生长行为和侵袭潜能。将6个细胞系接种于深度上皮化的大鼠气管（5 × 10^5个细胞/气管）。细胞接种后，将气管密封，移植到裸鼠皮下。在平行实验中，这些细胞系（1 × 10^6个细胞）注射s.c。在皮下，肿瘤成形率仅为13%，潜伏期长达18周，其中3个细胞系未表现出对周围组织的侵袭性生长。在大鼠气管中，所有细胞系均在3周内增殖，其中4个细胞系在1-2周内向气管壁浸润生长。气管壁上生长的细胞胸苷标记指数（^3H）高于气管管腔，细胞核更大，核仁突出，具有更强的异型性。气管癌的发生率与气管壁浸润性生长的发生率相关。原来携带六种肿瘤的患者的生存也与该系统的侵袭潜力密切相关。在气管移植中，所有细胞系的分化程度与原发病变相当或更高。而大多数细胞系在小鼠皮下的分化程度较低。气管移植物的表面重新填充鳞状细胞癌的细胞，最初有一层2至3个细胞厚的扁平细胞。而用3例腺癌细胞重新填充的细胞，最初内衬分别由立方体、圆形和柱状细胞组成的单层上皮。之后，这些细胞系分别呈现假层状、乳头状和筛网状生长模式。超微结构上，它们的结构分别类似纤毛支气管上皮细胞、非纤毛细支气管上皮细胞和支气管杯状细胞。这些结果表明，采用低传代细胞系的系统可以在接种后不久评估其侵袭潜力；细胞数量相对较少，可作为临床可靠的生物入侵试验。并且，使用该系统对培养细胞进行组织学检查被认为有利于评估分化，因为它提供了在原发性肿瘤或皮下直接细胞注射后形成的肿瘤中未观察到的独特增殖模式。少