THE ANALYSIS OF CHROMOSOMAL GAINS AND LOSSES IN PRIMARY MALIGNANTMELANOMA-COMPARISON WITH SPITZ NEVUS-

斯皮茨痣是一种良性黑素细胞肿瘤，临床和组织病理学检查很难或不能与黑色素瘤鉴别。我们尝试用比较基因组杂交（CGH）技术，在石蜡包埋的石蜡包埋组织中，从黑色素瘤中寻找染色体的增益和损失，并与Spitz痣的染色体进行比较。然而，从福尔马林固定、石蜡包埋的组织中提取的DNA的大小对于CGH来说太小了。因此，我们首先尝试用黑色素瘤细胞系MMG 1建立CGH方法。从正常雌性外周血淋巴细胞中提取参考DNA。我们得到了以下证据。DNA的提取：DNA Extractor WB Kit（和子，OSAKA）可用于细胞系的提取。2.标记时间：MMG 1细胞为一个半小时。三个小时的外周血淋巴细胞。乙醇沉淀时间：-80℃，1小时4。混合时间：37℃，3天，我们已经建立了CGH方法在我们的实验室，最后的问题是从福尔马林固定，石蜡包埋组织中提取DNA。我们将很快解决这个问题，我们可以通过比较基因组杂交（CGH）在福尔马林固定的石蜡包埋的黑色素瘤和斯皮茨痣组织中发现染色体的增益和损失。

SPITZ NEVUS IS A BENIGN NEOPLASM OF MELANOCYTES THAT CAN BE DIFFICULT OR IMPOSSIBLE TO DISTINGUISH FROM MELANOMA BY CLINICAL AND HISTOPATHOLOGIC EXAMINATION. WE TRIED TO FIND CHROMOSOMAL GAIN AND LOSSES BY COMPARATIVE GENOMIC HYBRIDIZATION (CGH) IN FORMALIN-FIXED, PARAFFIN ENMBEDDED TISSUES FROM MELANOMA, AND COMPARE WITH THAT FROM SPITZ NEVUS. HOWEVER, THE SIZE OF DNA EXTRACTED FROM FORMARIN-FIXED, PARAFFIN-EMBEDDED TISSUE IS TOO SMALL FOR CGH. THEREFORE, WE FIRST TRIED TO ESTABLISH THE CGH METHOD USING MELANOMA CELL LINE, MMG1. REFERENCE DNA WAS EXTRATED FROM NORMAL FEMALE PERIPHERAL BLOOD LYMPHOCYTES. WE GOT THE FOLLOWING RESULTS.1. EXTRACTION OF DNA : DNA EXTRACTOR WB KIT (WAKO, OSAKA) CAN BE USED FOR THE EXTRACTION FROM THE CELL LINES.2. LABELING TIME : ONE AND A HALF HOUR FOR MMG1 CELLS. THREE HOURS FOR PERIPHERAL BLOOD LYMPHOCYTES.3. PRECIPITATION TIME WITH ETHANOL : -80℃, ONE HOUR4. HYBRIDIZATION TIME: 37℃, 3 DAYSWE HAVE ESTABLISHED THE CGH METHOD IN OUR LABORATORY THE FINAL PROBLEM IS THE DNA EXTRACTION FROM FORMARIN-FIXED, PARAFFIN-EMBEDDED TISSUES. WE WILL SOLVE THE PROBLEM SOON AND WE CAN FIND CHROMOSOMAL GAIN AND LOSSES BY COMPARATIVE GENOMIC HYBRIDIZATION (CGH) IN FORMALIN-FIXED, PARAFFIN ENMBEDDED TISSUES FROM MELANOMA AND SPITZ NEVUS.