MOLECULAR BIOLOGICAL RESEARCH ON THE PRECURSOR AND PRECANCEROUS LESIONS OF THE KIDNEY-ANALYSIS WITH LASER CAPTURE MICRODISSECTION AND CHROMOGENIC IN SITU HYBRIDIZATION-

背景与目的：众所周知，von Hippel-Lindaus （VHL）病和持续血液透析后的肾脏含有肾细胞癌（RCC）的各种前体和癌前病变。在这里，我们的目的是利用激光捕获显微解剖（LCM）和显色原位杂交（CISH）来研究这种病变的遗传异常。材料和方法：对2例VHL病和7例持续性血液透析患者的肾脏组织，采用LCM切除非典型小管和囊肿，RT-PCR后进行cDNA微阵列分析，CISH评估与rcc发病机制相关的3、7、17和Y染色体数目。结果与讨论：提取的rna在质量上不适合RT-PCR或cDNA芯片。CISH也没能发现信号。与此同时，我们利用冷冻卵巢癌组织进行了LCM研究，以评估泌乳素受体的表达，并证明Her2基因在乳腺外Paget病中的拷贝数，这两项研究都取得了成功（Asai-Sato， Nagashima等人，in press; Ogawa， Nagashima等人，提交）。基于这些事实，在未来的研究中，碾压混凝土样品的采集方法需要重新考虑。

Back ground and aim: It is well known that the kidney of von Hippel-Lindaus (VHL) disease and after persistent hemodialysis contains various precursor and precancerous lesions of renal cell carcinoma (RCC). Here, we aimed to investigate the genetic abnormalities in such lesions using laser capture microdissection (LCM) and chromogenic in situ hybridization (CISH). Materials and methods: On the kidney tissue derived from 2 cases of VHL disease and 7 cases of persistent hemodialysis, LCM was performed to cut out atypical tubules and cysts for RT-PCR followed by cDNA microarray, and CISH for evaluate the cchromosome numbers of #3, 7, 17 and Y, which have been known to be related with pathogenesis of RCCs. Results and discussion: The extracted RNAs were not appropriate for RT-PCR or cDNA microarray in the quality. CISH also failed to reveal the signals. Along with these attempts, we performed LCM study using frozen ovarian cancer tissue to evaluate the expression of the prolactin receptor and to demonstrate the copy numbers of Her2 gene in extramammary Paget disease, both of which were successful (Asai-Sato, Nagashima et al., in press; Ogawa, Nagashima, et al., submitted). Based on these facts, the methods to collect the RCC samples should be reconsidered for future study.