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

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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.