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同一期的肿瘤经过相同的治疗,其结果可以显著不同,这可能是由于肿瘤的生物异型性所致。近年来流式细胞仪(FC)已被广泛用作发现这种异型性的有价值的工具,其取材多数为石腊包埋标本。作者对可以切除的非小细胞型肺癌(NSCLC)和胃肠道腺癌采取多处新鲜标本作DNA含量FC分析。病人的选择:共选用152例病人作为研究对象,计非小细胞型肺癌59例、胃癌38例和结肠直肠癌55例。术后均未给化疗,但17例直肠癌病人给予盆腔放疗。每6个月检查病人一次,随访5～10年。取样方法和制备;从肿瘤的中央和周边取2～4块新鲜标本,也包括肿大的淋巴结。样本置于RPMI1640培养液中,加入5%胎牛血清在4℃中贮存。后经剪碎后室温下在0.5%胃蛋白酶溶液中振动孵育10分钟进行酶学分离,用磷酸盐缓冲液洗去胃蛋白酶,过滤和离心,弃去上清液,细胞重新溶于磷酸盐缓冲液中经70%乙醇固定,加入溴化乙锭液和Mythramy-cin混合液中4℃贮存过夜。作FC分析,每一标本至少分析15000个细胞,算出DNA指数(DI)。统计学分析:采用生物医学数据处理统计软件
The same tumors undergo the same treatment and the results can be significantly different. This may be due to the tumor’s biological atypicality. In recent years, flow cytometry (FC) has been widely used as a valuable tool for the discovery of this type of atypicality, most of which are derived from paraffin-embedded specimens. The authors used multiple fresh specimens for resectable non-small cell lung cancer (NSCLC) and gastrointestinal adenocarcinoma for DNA content FC analysis. Patient selection: A total of 152 patients were selected as study subjects, including 59 non-small cell lung cancers, 38 gastric cancers, and 55 colorectal cancers. No chemotherapy was given after surgery, but 17 patients with rectal cancer were given pelvic radiotherapy. The patient was examined every 6 months and followed up for 5 to 10 years. Sampling methods and preparation; 2 to 4 fresh specimens from the center and periphery of the tumor, including enlarged lymph nodes. Samples were placed in RPMI 1640 medium and stored at 4°C with 5% fetal bovine serum. After being minced, the cells were enzymatically separated by shaking in a 0.5% pepsin solution at room temperature for 10 minutes. The pepsin was washed with phosphate buffer, filtered and centrifuged. The supernatant was discarded and the cells were redissolved in phosphate buffer. The solution was fixed with 70% ethanol and added to a mixture of ethidium bromide and Mythramy-cin at 4°C overnight. For FC analysis, analyze at least 15,000 cells per specimen and calculate the DNA index (DI). Statistical analysis: using biomedical data processing and statistical software