Böttger T, Gabbert H, Stöckle M, Schernos B, Junginger T
Klinik und Poliklinik für Allgemein- und Abdominalchirurgie, Johannes Gutenberg-Universität, Mainz, Bundesrepublik Deutschland.
Langenbecks Arch Chir. 1991;376(3):158-62. doi: 10.1007/BF00250341.
In this study we analysed DNA-ploidy as a potential prognostic parameter in papillary thyroid carcinoma. Paraffin embedded histological material, obtained by resection from 19 patients with a papillary thyroid carcinoma, was selected for analysis. Tumor areas within the paraffin-embedded material were identified by HE-stained reference sections. One 50 microns section was dewaxed, rehydrated and mechanically and enzymatically prepared to form a suspension of 10,000 cells/ml. 1 ml of the suspension, which contained bare nuclei with small rests of cytoplasma, was centrifuged on glass slides. The fixed nuclei were air-dried and stained by Feulgen SITS technique, which allows for the quantitative measurement of DNA. The DNA analysis was carried out with a computer-controlled single-cell cytophotometry. In contrast to using flow cytometry, only the tumor cells were measured by image-cytometry. Overlapping nuclei, dirt and other artifacts as well as inflammatory cells were efficiently eliminated. With DNA image-cytometry, we could differentiate between diploid (n = 13) and aneuploid (n = 6) tumors. Best prognosis with a survival rate of 92% after 103 months had patients with diploid tumors in contrast to patients with aneuploid tumors who did not survive more than 72 months.
在本研究中,我们分析了DNA倍体作为甲状腺乳头状癌潜在预后参数的情况。选取了19例甲状腺乳头状癌患者手术切除获得的石蜡包埋组织学材料进行分析。通过苏木精-伊红(HE)染色的参照切片确定石蜡包埋材料中的肿瘤区域。将一张50微米厚的切片脱蜡、水化,经机械和酶处理制成每毫升含10000个细胞的悬液。取1毫升含有裸核及少量细胞质残余的悬液,离心至载玻片上。固定后的细胞核经空气干燥,采用福尔根-席夫(Feulgen SITS)技术染色,该技术可对DNA进行定量测量。DNA分析通过计算机控制的单细胞细胞光度测定法进行。与使用流式细胞术不同,图像细胞术仅测量肿瘤细胞。重叠的细胞核、污垢及其他伪像以及炎性细胞被有效去除。通过DNA图像细胞术,我们能够区分二倍体肿瘤(n = 13)和非整倍体肿瘤(n = 6)。二倍体肿瘤患者预后最佳,103个月后的生存率为92%,而非整倍体肿瘤患者生存时间不超过72个月。