Experimental Study of Metastasis and Invasion Related Oncogene by Using Oral Cancer Cell Lines

假设癌细胞在细胞外分解基质是转移所必需的，而基质金属蛋白酶(以下简称基质金属蛋白酶)的表达对癌细胞的转移非常重要。有报道称，Ets结合Domein位于这些基因上游的转录调控区，通过结合Ets癌基因家族产物，在这些基因的转录调控中发挥重要作用。部分细胞株有E1AF蛋白表达，但无强蛋白表达细胞株。也就是说，有许多癌细胞E1AF mRNA表达较强，但未见E1AF蛋白表达较强的癌细胞。因此，我们怀疑存在E1AF蛋白表达非常低或…降解的可能性由于癌细胞的低稳定性，很快就会变得更敏感。将逆转录病毒增强子/启动子下的E1AF基因导入不表达E1AF基因的癌细胞，建立产生细胞系。裸鼠皮下注射这些细胞后，其转移能力和运动能力均有明显提高。这些数据支持了E1AF基因对恶性肿瘤的转移和侵袭有很大影响的假说。同时还对MMP3和MMP9等基质金属蛋白酶蛋白的表达进行了分析。我们调查了这些基因的表达、肿瘤的组织病理类型(恶性程度)和预后，并分析了它们之间的关系。一般认为E1AF基因的蛋白表达越强，肿瘤越恶性(预后越差)。(MM3、MMP9等基质金属蛋白酶蛋白同时也高表达)，这些基因的表达与肿瘤的恶性程度和转移能力密切相关。较少

It is assumed that decomposing matrix outside the cell is necessary for the metastasis of the cancer cells, and the expression of the matrix metaroprotease (henceforth MMP) is important for it. It had been reported that the ETS binding domein was located in the transcriptional regulation area in the upstream of these genes, and which played an important role in the transcriptional regulation of these genes by binding of ets-oncogene family products.The protein was extracted from oral malignant tumors (malignant squamous cell carcinoma cell lines), and the expressions of the E1AF protein were studied by the Western-blot analysis. There were some cell lines in which E1AF protein was expressed, but no cell lines of strong protein expression. In other words, there were many cancer cells in which E1AF mRNA was strongly expressed, but no cancer cells of strong protein expression of E1AF. So, we suspected that there was some possibility that the E1AF protein expression was very low or decompo … More sed soon because of that low stability in cancer cells. The E1AF gene under the retrovirus enhancer/promoter was transfected into the cancer cells in which E1AF was not expressed, and the producer cell lines were established. The abilities of metastasis and mobility were elevated in the experiment of injecting these cells to nude mouse subcutaneously. These data support the hypothesis that the metastasis and invasion of a malignant tumor had deeply been affected by E1AF gene.The analysis of the expressions such as the MMP proteins (MMP3 and MMP9 proteins) was also carried at the same time. We investigated the expressions of these genes, the histopathological type of cancer (malignant degree), and the prognosis, and the relation between them was also examined.It was generally thought that the stronger protein expression of E1AF gene was, the more malignant the cancer was (the worse prognosis was). (The expressions of the MMP proteins such as MM3 and MMP9 protein were also high at the same time), and the expressions of these genes have much relation to the malignant degree of cancer and the metastasis ability in many cases of cancer cell lines. Less