Immunohistological study for the invasion and metastasis of oral malignant melanoma cell line (SSM-1)

侵袭性和转移性是口腔恶性黑色素瘤最重要的特征。为了阐明口腔恶性黑色素瘤侵袭性和转移性表达的影响因素，我使用了具有三维I型胶原凝胶基质的高黑色素生成细胞系（SSM-1）。该方法可用于获得体外条件。实验结果如下：1。2-3周后SSM-1融合。样品用10%福尔马林固定，石蜡包埋。石蜡切片用H-E染色。SSM-1的形状为纺锤形和多边形。我尝试了两种体外入侵模型的方法。(1)将人成纤维细胞凝胶与SSM-1菌落混合(2)人成纤维细胞在包埋胶原凝胶基质中培养后，将SSM-1覆盖在凝胶基质上。用4%多聚甲醛固定样品，进行抗人E-cadherin抗体免疫组化染色。用SAB法对切片进行钙粘蛋白免疫染色。SSM-1和原发病变活检未染色。结果表明，E-cadherin表达的降低表明细胞间黏附能力的降低。rt - pcr检测MMP-2。SAB法对SSM-1和原发病变活检组织进行MMP-2 - MMP9免疫染色。由于黑素细胞的存在，原发病灶未检测到mmp -2,9。在SSM-1中，MMP-9未染色，但在成纤维细胞中也有MMP-2染色。这些结果表明成纤维细胞与侵袭和转移能力密切相关。

The invasive and metastatic ability are most important characteristics of oral malignant melanoma. In order to elucidate the factors contributory to the expression of invasiveness and metastasis of oral malignant melanoma, I used high melanin productive cell line(SSM-1) with three-dimensional type I collagen gel matrix. This method is usefull to obtain the condition of iv vivo. The results were folloewd :1. SSM-1 was confluent after 2-3weeks.2. The samples were fixed with 10% formalin, and embedded in paraffin. Paraffin sections were stained with H-E.The shape of SSM-1 were spindle and polygonal.3. I tried two method for an in vitro invasion model. (1) mixed the gel of human fibroblast with the colony of SSM-1 (2)after incubated human fibroblast in the fibroblast-embedded collagen gel matrix, overlaid SSM-1 on the gel matrix.4. The samples were fixed with 4% paraformaldehyde for immunohistochemical stain of anti-human E-cadherin antibody. The sections were immunostained for cadherin by SAB method. SSM-1 and the biopsy of primary lesion were not stained. This result suggest that the decrease of E-cadherin expression indicated the decrease of cell-cell adhesion.5. MMP-2 was detected with RT-PCR.6. SSM-1 and the biopsy of primary lesion were immunostained for MMP-2 MMP9 by SAB method.Because of melanocyto, I could not detect MMP-2,9 in primary lesion. In SSM-1, MMP-9 was not stained, but MMP-2 was stained in the fibroblast too.These results suggest that fibroblast is well correlated with the invasive and metastatic ability.