The basic research for characterization of AMF/AMFR and application to diagnosis.

AMF/AMFR表征及其诊断应用的基础研究。

• 批准号: 06557108
• 负责人: AMAGASA Teruo
• 金额: $ 6.98万
• 依托单位: Faculty of Dentistry, Tokyo Medical and Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06557108/
• 关键词: AMF; Squamous cell carcinoma; Boyden chamber; Northern blot analysis; RT-PCR; TNM classification; AMFR; invasion and metastasis; ボイデン・チェンバー; 自己分泌型遊走因子; 分子生物学; クローニング; レセプター; 細胞工学

AMF (Autocrine Motility Factor) has been reported to be produced by malignant melanoma, fibrosarcoma and bladder carcinoma cells, with highly invasive and metastatic potentials. Boyden chamber analyzes showed that conditioned medium of oral squamous cell carcinoma (SCC) LMF4 cells also induced cell motility in an autocrine fashion. This motile activity was purified by using molecular sieve column chromatography and DEAE high performance liquid chromatography, and was identified as a single protein (LMF4 AMF) with molecular weight of 55kD and 65kD under non-reduced and reduced condition, respectivetly. The LMF4-AMF also induced fibrosarcoma cell motility in a dose-dependent manner as well as other SCC cells. These results suggested that LMF4-AMF was identical to other AMFs reported previously.The LMF4 cell motility was also induced by the AMF produced by fibrosarcoma cells, suggesting that LMF4 cells expressed functional AMFR (AMF Receptor). Northern blot and RT-PCR analyzes of SCC cells showed that invasive and metastatic cells expressed higher amount of AMFR than non invasive SCC cells. RT-PCR analyzes of clinical specimens showed that prinary cells expressed AMFR in relation to N stages rater than T in TNM classification and that metastasized cells expressed AMFR more than primary.We conclude that SCC also produced AMF and express AMFR and that these molecules are related to SCC cell motility in association with invasive and metastatic potentials of SCC cells.

据报道，AMF（自分泌运动因子）由恶性黑色素瘤、纤维肉瘤和膀胱癌细胞产生，具有高度侵袭性和转移潜力。Boyden小室分析表明，口腔鳞状细胞癌（SCC）LMF 4细胞的条件培养基也诱导细胞运动的自分泌方式。经分子筛柱层析和DEAE高效液相色谱纯化，鉴定为一个单一的蛋白质（LMF 4 AMF），在非还原和还原条件下的分子量分别为55 kD和65 kD。LMF 4-AMF还以剂量依赖性方式诱导纤维肉瘤细胞以及其他SCC细胞的运动。这些结果表明，LMF 4-AMF与以往报道的AMF相同，LMF 4细胞的运动也可被纤维肉瘤细胞产生的AMF所诱导，表明LMF 4细胞表达功能性AMFR（AMF受体）。北方杂交和RT-PCR分析显示，浸润和转移的SCC细胞中AMFR的表达高于非浸润的SCC细胞。RT-PCR分析表明，原发癌细胞表达AMFR与TNM分期的关系比T分期的关系更密切，转移癌细胞表达AMFR也高于原发癌细胞，提示SCC也产生AMF和表达AMFR，这些分子与SCC细胞的运动性有关，并与SCC细胞的侵袭和转移能力有关。

项目成果

新中康史,天笠光雄: "基底膜浸潤とインテグリンα6β4" 日本口腔組織培養研究会雑誌. 4. 11-19 (1995)

Yasushi Shinnaka、Mitsuo Amakasa：“基底膜侵入和整合素α6β4”日本口腔组织培养研究会杂志4. 11-19 (1995)。

Narikazu Uzawa: "Transter of a normal chromosome 3 Suppresses Tumori-genisity of Oral squamous cell carcinoma cell lines" Oral Oncology. 4B. 265-268 (1995)

Narikazu Uzawa：“正常 3 号染色体的转移抑制口腔鳞状细胞癌细胞系的肿瘤发生”口腔肿瘤学。

Masao Saitoh: "Identification of important regions in the cytoplaimic ynxtamenbrane donain of typa 1 receptor that separate sigvaling pathmays of transforming growth factor-β" The Jourral of Biological Chemistry. 271. 2769-2775 (1996)

Masao Saitoh：“Typa 1 受体的细胞质膜膜区中分离转化生长因子-β 信号通路的重要区域的鉴定”《生物化学杂志》271. 2769-2775 (1996)。

新中康史: "基底膜の認識・接着に関与する扁平上皮癌細胞表面分子の研究" 日本口腔組織培養研究会雑誌. 4. 93-94 (1995)

Yasushi Shinnaka：“参与基底膜识别和粘附的鳞状细胞癌细胞表面分子的研究”日本口腔组织培养研究会杂志4. 93-94（1995）。

Masafumi Mimura, Nobuyuki Tanaka, Takao Miyamoto, Ken-ichi Shionoya, Yutaka Kimishima and Teruo Amagasa: "Sensitivity of malignant melanoma line cells derived from oral mucosa to radiation" Jpn.J.Tissue Cult.Dent.Res.4. 113-114 (1195)

Masafumi Mimura、Nobuyuki Tanaka、Takao Miyamoto、Ken-ichi Shionoya、Yutaka Kimishima 和 Teruo Amagasa：“源自口腔粘膜的恶性黑色素瘤系细胞对辐射的敏感性”Jpn.J.Tissue Cult.Dent.Res.4。