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目的建立一种循环肿瘤细胞的检测方法,探讨肝癌患者外周血中循环肿瘤细胞在肿瘤进展中的意义。方法肝癌细胞株HCC97-L经滤过富集后测定肿瘤细胞的检出率以检测该方法的敏感性,采用该方法滤过富集32例肝细胞癌(HCC)患者、15例肝良性病变患者和10例正常健康对照的外周血中循环肿瘤细胞,用细胞角蛋白8/18(CK8/18)行免疫荧光检测结合显微镜下形态学观察,判断并计数肿瘤细胞。结果HCC97-L细胞检出率在40%～100%之间,平均为68%;Ⅰ～Ⅱ期患者,循环肿瘤细胞的阳性率为28.6%;Ⅲ～Ⅳ时,为88.9%。两组差异有统计学意义(P<0.01)。健康志愿者10例和肝良性病变患者15例外周血均未见CK8/18阳性肿瘤细胞。结论全血滤过富集法能简单、快速、有效的检出肝癌患者外周血中循环肿瘤细胞,部分早期肿瘤已有肿瘤细胞的血源性播散;随着肿瘤进展血中肿瘤细胞也增加。
Objective To establish a detection method of circulating tumor cells and explore the significance of circulating tumor cells in the peripheral blood of patients with liver cancer. Methods The detection rate of tumor cells in HCC97-L hepatoma cell line HCC97-L was determined by enrichment. The sensitivity of the method was tested by this method. Enrichment of 32 HCC patients and 15 benign liver lesions Tumor cells were harvested from the peripheral blood of 10 healthy volunteers and 10 healthy controls. Cytokeratin 8/18 (CK8 / 18) immunofluorescence assay was used to observe the morphological changes of the tumor cells. Results The detection rate of HCC97-L cells was between 40% and 100% with an average of 68%. The positive rate of circulating tumor cells was 28.6% in stage Ⅰ ~ Ⅱ patients and 88.9% in stage Ⅲ ~ Ⅳ. The difference between the two groups was statistically significant (P <0.01). CK8 / 18 positive tumor cells were found in 10 healthy volunteers and 15 patients with benign liver lesions. Conclusion The whole blood filtration enrichment method can detect the circulating tumor cells in the peripheral blood of patients with liver cancer easily, rapidly and effectively. Some of the early tumors have had the blood-borne dissemination of the tumor cells. As the tumor progresses, the number of tumor cells in the blood also increases .