Department of Pathology, Institute of Basic Medical Sciences and School of Basic Medicine, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100005, China.
J Cancer Res Clin Oncol. 2012 Oct;138(10):1651-8. doi: 10.1007/s00432-012-1242-y. Epub 2012 May 24.
Malignant melanoma, characterized by early distant metastasis to the lungs and brain, is a leading cause of mortality related to skin cancer. Cell fusion and the subsequent aneuploidy, commonly observed in melanoma, are associated with poor prognosis. However, the pathological consequences of cell fusion in melanoma remain unknown. Therefore, the present study aims to investigate the pathological consequences of cell fusion in melanoma and the mechanism of melanoma metastasis.
Phytohemagglutinin-polyethylene glycol (PHA-PEG) fusion method was developed for the fusion of tumor cells. Melanoma cells were fused through the improved PHA-PEG fusion method and obtained by fluorescence-activated cell sorting. DNA content was analyzed through flow cytometry. Cell proliferation rate was detected by cell culture in vitro, and the cell number was counted daily. To detect the tumor growth rate in vivo, cells were injected subcutaneously and the tumor volumes were measured using a vernier caliper. To analyze the tumor metastatic potential, cells were injected intravenously, and the collected lung-metastasis samples were weighed by an electronic balance and the surface nodules were counted.
We established an improved phytohemagglutinin-polyethylene glycol fusion method and successfully obtained stable melanoma tumor-tumor cell fusion hybrids. Cell size, DNA content, and chromosome numbers of the fusion hybrids were approximately twice those of the parents. The metastatic potential of the fusion hybrids was dramatically enhanced, in contrast to their proliferation rate. Their metastasis was specific to the lungs.
We developed a highly efficient cell fusion method that can be applied in many fields, particularly cancer research. Our study has proven that tumor-tumor cell fusion hybrids in melanoma can acquire enhanced and specific metastatic potential. Thus, blockage of cell fusion may be a new strategy for melanoma metastasis therapy.
恶性黑色素瘤以早期肺和脑远处转移为特征，是导致皮肤癌相关死亡的主要原因。细胞融合和随后的非整倍体，在黑色素瘤中常见，与预后不良相关。然而，黑色素瘤中细胞融合的病理后果尚不清楚。因此，本研究旨在探讨黑色素瘤中细胞融合的病理后果及黑色素瘤转移的机制。
采用植物血凝素-聚乙二醇（PHA-PEG）融合法进行肿瘤细胞融合。通过改良的 PHA-PEG 融合法融合黑色素瘤细胞，通过荧光激活细胞分选获得。通过流式细胞术分析 DNA 含量。通过体外细胞培养检测细胞增殖率，每天计数细胞数。为了检测体内肿瘤生长率，将细胞皮下注射，并使用游标卡尺测量肿瘤体积。为了分析肿瘤转移潜能，将细胞静脉内注射，通过电子天平称重收集的肺转移样本，并计数表面结节。
我们建立了一种改良的植物血凝素-聚乙二醇融合方法，并成功获得了稳定的黑色素瘤肿瘤-肿瘤细胞融合杂种。融合杂种的细胞大小、DNA 含量和染色体数量大约是亲本的两倍。与增殖率相比，融合杂种的转移潜能显著增强。它们的转移是针对肺部的。
我们开发了一种高效的细胞融合方法，可应用于许多领域，特别是癌症研究。我们的研究证明，黑色素瘤中的肿瘤-肿瘤细胞融合杂种可以获得增强的特异性转移潜能。因此，阻断细胞融合可能是一种治疗黑色素瘤转移的新策略。
