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

Spitz痣是一种黑素细胞良性肿瘤，通过临床和组织病理学检查很难或不可能与黑色素瘤相鉴别。我们试图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组织中发现染色体的得失，并与Spitz痣组织进行比较。然而，从FORMARIN固定的石蜡包埋组织中提取的DNA对于CGH来说太小了。因此，我们首先尝试利用黑色素瘤细胞系MMG1建立CGH方法。从正常女性外周血淋巴细胞中提取参比DNA。我们得到了以下结果。DNA的提取：DNA提取试剂盒(WAKO，大阪)可用于从LINES2细胞中提取DNA。标记时间：MMG1细胞一个半小时。外周血淋巴细胞检测3小时。乙醇沉淀时间：-80℃，1HUR4。杂交时间：37℃，3 DAYSWE在我们实验室建立了CGH方法。最后一个问题是从福尔马林固定的石蜡包埋组织中提取DNA。我们很快就会解决这个问题，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兹痣组织中发现染色体的得失。

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.