Prediction of minute prostatic cancer development by telomere length measured with tissue section FISH method.

2005年和2006年，我们两次邀请加拿大BC癌症中心的Steven Poon博士作为我们研究团队的一员。我们与潘博士合作，设计了一个新的软件程序，“Tissue Telo version 2”，用于使用组织切片测量端粒长度。利用这种方法，并使用荧光原位杂交（FISH）与端粒特异性探针在组织切片上，我们能够测量端粒长度的前列腺组织的档案切片，并获得了以下数据：1.前列腺组织分析的困难前列腺组织包含粗糙的肌肉和胶原纤维，它们具有非常强的自发荧光。这种自发荧光使我们无法测量上皮和间质中的端粒长度。尽管尝试了几种去除这种自发荧光的方法，但不能获得足够令人满意的荧光显微镜图像。2.荧光原位杂交定量方法的改进以着丝粒荧光强度为内对照，端粒长度以端粒/着丝粒强度比（TCR）表示。此外，我们使用了TIG-1细胞块切片的TCR，以获得更精确的数据。标准化TCR是组织切片的TCR除以来自细胞块的TIG-1切片的TCR。3.癌细胞的TCR是正常成纤维细胞的71-83%。根据这些不同的TCR，区分癌细胞和成纤维细胞似乎是可行的。4.前列腺中（腔细胞）的TCR小于（基底细胞）的TCR。提示前列腺组织中管腔细胞和基底细胞构成不同的细胞群。5.正常前列腺成纤维细胞TCR与癌成纤维细胞TCR不同。这表明，这些细胞群在基质中是不同的。

We invited Dr. Steven Poon from the BC Cancer Center, Canada, twice in 2005 and 2006, to work as a member of our research team. In collaboration with Dr. Poon, we devised a new software program, "Tissue Telo version 2", for measurement of telomere length using tissue sections. With this approach, and the use of fluorescence in situ hybridization (FISH) with a telomere-specific probe on tissue sections, we were able to measure telomere length in archival sections of prostatic tissue, and obtained the following data :1.Difficulties with analysis of prostatic tissueProstatic tissue contains rough muscle and collagen fibers, which have very strong autofluorescence. This autofluorescence prevented us from measuring telomere length in epithelium and stroma. Although several methods for removing this autofluorescence were tried, sufficiently satisfactory fluorescence microscopy images could not be obtained. Thus it is more difficult to obtain good images using prostate than with gastrointestinal tissue.2.Improvement of quantitative fluorescence in situ hybridizationWe used centromere fluorescence intensity as an internal control, and expressed telomere length as the telomere/centromere intensity ratio (TCR). Moreover, we used TCR of cell block sections of TIG-1 to obtain more precise data. The normalized TCR was the TCR of a histologic section divided by the TCR of a section of TIG-1 from a cell block.3.TCRs of carcinoma cells were 71-83% those of normal fibroblasts. On the basis of these different TCRs, it appeared feasible to distinguish cancer cells from fibroblasts.4.TCRs of (luminal cells) were smaller than those of (basal cells) in the prostate gland. It is suggested that luminal and basal cells constitute different populations in the prostate.5.TCRs of normal fibroblasts were different from those of cancer fibroblasts. It is suggested that these cell populations in the stroma are different.