Application of Modified FISH to Analyze Tumor Cell Radioresistance and Development of Its Clinical Application

在常规的染色体分析中，处于M期的细胞可以作为研究对象。然而，这种方法的效率很低，无法得到足够的结果。以染色体转化为指标时，该方法的灵敏度也较低。因此，染色体畸变素的分析一直是一项艰难的工作。本研究用荧光染料对浓缩后的染色体进行染色，代替常规染色方法。1999年用集落试验检测了膀胱癌、卵巢癌、子宫颈癌、食道癌培养细胞的放射敏感性，获得了细胞放射敏感性的基础数据。结果表明，不同DO值的培养细胞对辐射的双重敏感性不同。2000年对受照淋巴细胞的染色体畸变率进行了检测。采集全身照射患者各照射剂量(2~12Gy)的淋巴细胞。用染色体分析和1、2号染色体FISH分析了染色体畸变率与辐射剂量的关系，结果表明染色体畸变率与辐射剂量有良好的相关性。对同一患者采集的淋巴细胞进行体外照射，染色体畸变率与照射剂量的相关性与体内分析结果一致。这一结果表明，在2-12Gy剂量范围内，用FISH技术分析1、2号染色体的体外淋巴细胞染色体畸变率与体内染色体畸变率有很好的相关性。在对肿瘤细胞的染色体畸变率分析中，发现照射前细胞发生了较大的突变。因此，很难评估辐射的效果。这项研究将在未来继续对肿瘤细胞进行。

In the conventional chromosome analysis, the cells in the M-phase can be the target. However, the efficiency of this method was very low and sufficient results could not be expected. When the transformation of the chromosome was adapted as the index, the sensitivity of this method was also low. As a result, analysis of chromosome aberratoin had been a tough business. The condensed chromoses were stained with fluorescent dyes for the specific chromosome in this study instead of the conventional method. The radiosensitivity of the culture cells established from bladder cancer, ovarian cancer, uterine neck cancer, esophageal cancer were examined by the Colony assay in 1999, and basic data for radiosensitivity of the cells were obtained. As a results, the culture cells of which differed in the Do value of about the double radiosensitivity were clarified. Chromosomal aberration in irradiated lymphocytes was examined in 2000. The lymphocytes were collected at each exposure radiation dose(2-12 Gy)from the patient with the whole body irradiation. The relationship between chromosomal aberration and exposure radiation dose was analyzed with chromosome analyses and FISH at chromosome 1, 2. As The result, there was good correlation between the chromosome aberrations and irradiation doses. When the lymphocytes collected from the same patient were irradiated in in vitro, the same correlation between chromosome aberrations and irradiation doses was found as the in vivo analyses. This result suggests that chromosome aberration of lymphocytes analysed in vitro with FISH at chromosome 1, 2 has a good correlation with that in vivo at the doses between 2-12 Gy. In the analyses of chromosome aberration of the tumor cells, there were some much transformations before the irradiation. As a result, it was difficult to evaluate the effect of irradiation. This study will be continued with tumor cells in the future.