CAJ | 학술논문

目的改进和完善改良"彗星"分析法以更好地用于检测实体肿瘤的放射敏感性.方法门诊活检标本制成单细胞悬液,冰上照射后立即铺胶制板,在裂解液中裂解50 min,漂洗后进行细胞电泳20 min.PI染色后在荧光显微镜下采图并用专用软件进行图像分析.每例样品均设对照(包括外参对照和自身对照)+m5 Gy照射,并对活检样品的正常细胞和肿瘤细胞进行分类采图.观察指标为细胞DNA含量和尾力矩.结果影响实验室测定结果与临床肿瘤放射反应相关性的因素主要有:(1)取材较表浅样品中仅有淋巴细胞而无肿瘤细胞,从而导致实验室测定结果与临床肿瘤放射敏感性的吻合性下降;(2)样品中坏死细胞较多,使本底增高,是影响放射敏感性分析的重要因素;(3)增设淋巴瘤细胞作为外参对照可对实验系统误差进行校正,是质量控制不可缺少的环节;(4)对活检样品中正常细胞和肿瘤细胞进行分类采图能提高临床与实验室检测结果的吻合性.结论实验同时设置内、外参对照和对样品中肿瘤细胞和正常组织细胞进行分类采图,能不同程度提高实验室检测结果与临床肿瘤实际放射反应的吻和性.
목적 개진화완선개량"혜성"분석법이경호지용우검측실체종류적방사민감성.방법 문진활검표본제성단세포현액,빙상조사후립즉포효제판,재렬해액중렬해50 min,표세후진행세포전영20 min.PI염색후재형광현미경하채도병용전용연건진행도상분석.매례양품균설대조(포괄외삼대조화자신대조)+m5 Gy조사,병대활검양품적정상세포화종류세포진행분류채도.관찰지표위세포DNA함량화미력구.결과 영향실험실측정결과여림상종류방사반응상관성적인소주요유:(1)취재교표천양품중부유림파세포이무종류세포,종이도치실험실측정결과여림상종류방사민감성적문합성하강;(2)양품중배사세포교다,사본저증고,시영향방사민감성분석적중요인소;(3)증설림파류세포작위외삼대조가대실험계통오차진행교정,시질량공제불가결소적배절;(4)대활검양품중정상세포화종류세포진행분류채도능제고림상여실험실검측결과적문합성.결론 실험동시설치내、외삼대조화대양품중종류세포화정상조직세포진행분류채도,능불동정도제고실험실검측결과여림상종류실제방사반응적문화성.
Objective To impmve the method of "modified comet assay" in predicting the radiosensitivitv of solid tumor. Methods A single cell suspension from biopsy sample was lmdlated on ice with a dose of 5 Gy.The microscope slide was spread with agarose,lysed for 50 minutes,rinsed 3 times rinse solution,and given electrophoresis for 20 minutes. After being stained with PI,cell images were collected through the microscope and analyzed with Lucia G software(Version 4.6).In order to check system/ background errors,every sample was made into control slide and irradiation slide.The end-points were cell DNA contents and tail moment. Results The factors influencing the results included:(1)Sample was iaulty tor the biopsv taken from mucosa and no tumor cells were contained. (2)The slides with a high backgmund ( induced by necrosis) disturbed the measurement of comet assay. (3) Setting lymphocytes as control to check svstem errors was very important. (4)To separately collect images of the normal tissue cells and tumor cells from the biopsy sample improved the conformity between the clinical obscrvation and the lab result. Conclusions To increase the correlation between comet assay and clinical response,it is very helpful to set double control for checking system/background errors and to collect images of the normal tissue cells and tumor cells through the microscope,respectively.