A possible evaluation system for effect of oral cancer chemotherapy based on the grading of apoptotic changes in the carcinoma cells

为了检验口腔鳞状细胞癌化疗过程中癌细胞凋亡的分级变化是否有助于评估治疗效果，我们对化疗前后标本制备的组织切片中的凋亡癌细胞进行了检测。用末端脱氧核苷酸转移酶(TdT)-生物素末端标记法(TUNEL)检测新鲜冰冻组织切片中的凋亡细胞。虽然TUNEL法检测鳞癌中的凋亡细胞可作为评价肿瘤化疗效果的一种工具，但仅用TUNEL法检测凋亡细胞并不足以作为评价化疗效果的指标。此外，由于涉及复杂的过程和使用昂贵的试剂，TUNEL方法执行起来复杂且昂贵。作为原位检测细胞凋亡的另一种方法，单链…的免疫组织化学检测更多的ed(Ss)DNA已被证明对识别经历了细胞凋亡或程序性细胞死亡的细胞具有重要的价值。为了验证免疫组织化学方法的可靠性，我们比较了两种方法，即TUNEL法和单链DNA免疫组织化学方法对SCC细胞凋亡程度的评估，结果发现，这两种方法评估的细胞凋亡指数之间存在显著的相关性。然而，即使成功地进行了单链DNA的免疫组织化学检测，仅检测鳞状细胞癌中的凋亡细胞的评估系统仍然是不够的。因此，我们还研究了SCC细胞的额外细胞标记物，在这些标记物中表达可能受到癌症化疗的影响。结果提示，化疗前后鳞癌标本中E-钙粘蛋白、α连环蛋白、硫酸肝素糖胺聚糖、细胞周期蛋白D1和Ki67免疫组织化学检测的对比试验可作为评价肿瘤化疗效果的一个候选指标。较少

To test whether grading of apoptotic changes of carcinoma cells in the course of chemotherapy for oral squamous cell carcinoma (SCC) can be useful in evaluation of the effect of the therapy, we examined the apoptotic carcinoma cells in tissue sections prepared from specimens obtained before and after the chemotherapy. Apoptotic cells in the fresh frozen tissue sections were detected by terminal deoxynucleotidyl transferase (TdT)-biotin end-labeling (TUNEL) method. Although detection of apoptotic cells in the SCC by the TUNEL method might be useful as a tool to evaluate the effect of the cancer chemotherapy, examination of apoptotic cells by only the TUNEL method was not enough for the evaluation system for the effect of chemotherapy. The TUNEL methods, moreover, is complex and expensive to perform because of the complicated processes involved and the use of the expensive reagents. As an another method to detect apoptotic cells in situ, the immunohistochemical detection of single-strand … More ed (ss) DNA has demonstrated to be great value of identification of cells which undergo apoptosis or programmed cell death. To examine reliability of the assay by means of the immunohistochemistry, we comparatively investigated the extent of apoptotic cell loss in SCCs evaluated by two methods, namely the TUNEL method and immunohistochemistry for ssDNA.As a result, a significant correlation in the apoptotic indexes evaluated by these methods was found. However an evaluation system by only detection of apoptotic cells in SCC were still not enough even if immunohistochemistry for ssDNA was successfully performed. Thus we also investigated additional cell marker of SCC cells in which expression might be influenced by cancer chemotherapy. As a result, it was suggested that comparative test concerning the immunohistochemical detection of E-cadherin, α catenin, heparansulfate glycosaminoglycan, Cyclin D1, and Ki67 in the SCC specimens obtained before and after the chemotherapy might be a candidate evaluation system for the effect of the cancer chemotherapy. Less