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背景与目的:检测胃癌患者骨髓微转移有多种方法,但不同的方法差异很大。在乳腺癌中,以磁激活细胞分选(m agnetic activated cellsorting,M ACS)结合荧光激活细胞分类(fluorescentactivated cellsorting,FACS)检测血细胞中的肿瘤细胞,有较高的敏感性和特异性。本研究探讨应用这一方法检测胃癌骨髓微转移的临床意义。方法:选择2002年12月～2003年6月行手术治疗的胃癌患者35例,提取其骨髓有核细胞,以结合抗细胞角蛋白(cytokeratin,CK)7/8抗体的M ACS微型免疫磁珠、标记异硫氰酸荧光素(fluorescein isothiocyanate,FITC)的抗CK抗体,以及标记叶绿素蛋白的抗CD45抗体标记后,以M S+/RS+型阳性分选柱进行两次肿瘤细胞富集。取富集前后的细胞样本进行FACS检测,将检测结果与各临床病理学参数进行比较。结果:M ACS富集前,仅3例(8.6%)检测到骨髓中存在的微转移细胞;而富集后,有25例(71.4%)患者检测到了骨髓微转移细胞。组织学分级为中分化、低分化和未分化的患者中,肿瘤细胞频数分别为1.4×10-8～2.4×10-5、2.2×10-7～3.7×10-5和4.0×10-6～8.6×10-5,3组比较,有显著性差异(P=0.026)。骨髓微转移状况与肿瘤TNM分期密切相关(r=439,P=0.008),而与肿瘤大小、浸润深度等参数无关。结论:M ACS结合FACS可提高胃癌患者骨
Background and Objective: There are many ways to detect bone marrow micrometastases in patients with gastric cancer, but different methods vary greatly. In breast cancer, the detection of tumor cells in blood cells by using MACS (activated carbon cell sorting) combined with fluorescent activated cell sorting (FACS) has high sensitivity and specificity. This study was to explore the clinical significance of using this method to detect bone marrow micrometastases in gastric cancer. Methods: Thirty-five patients with gastric cancer who underwent surgical treatment from December 2002 to June 2003 were enrolled in this study. The bone marrow cells were obtained and combined with M ACS mini-immunomagnetic beads with anti-cytokeratin (CK) 7/8 antibody , Anti-CK antibody labeled with fluorescein isothiocyanate (FITC), and anti-CD45 antibody labeled with chlorophyll, and then twice enriched in tumor cells by M S + / RS + positive sorting column. Before and after enrichment of cell samples for FACS detection, the test results and the clinical pathological parameters were compared. RESULTS: Only three (8.6%) of the micro-metastatic cells in the bone marrow were detected prior to M ACS enrichment; however, bone marrow micrometastatic cells were detected in 25 (71.4%) patients after enrichment. Tumor cell frequencies were 1.4 × 10-8 ~ 2.4 × 10-5, 2.2 × 10-7 ~ 3.7 × 10-5 and 4.0 × 10-6 in patients with moderately differentiated, poorly differentiated and undifferentiated tissues, respectively ~ 8.6 × 10-5,3 group, there was a significant difference (P = 0.026). Micro-metastasis of bone marrow was closely related to tumor TNM stage (r = 439, P = 0.008), but not to parameters such as tumor size and depth of invasion. Conclusion: M ACS combined with FACS can improve the bone of patients with gastric cancer