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目的 筛选B16M高低转移株 ,以探讨B16M的转移异质性及机制。方法 通过B16M自发肺转移模型筛选出转移性能不同的细胞株 ,并用实验性和自发性肿瘤转移模型验证其在体内生长、转移性。通过细胞增殖、3 H TdR参入实验、流式细胞技术、细胞侵袭离散实验研究了高、低转移B16M细胞株体外增殖能力、侵袭能力以及对离散因子的反应的差异 ,Western印迹实验研究细胞c -met表达及活化水平。结果 在所筛选出的 3株可疑高转移B16M细胞株中 ,H3B16M细胞接种于同系C5 7BL/ 6小鼠体内所形成的原发瘤重量、实验和自发性肺转移结节数明显高于其他细胞株 ;该细胞株增殖能力、侵袭性及对离散因子的反应能力的也明显高于其他细胞株。c met表达及磷酸化水平也升高。结论 成功的筛选出B16M高、低转移细胞株 ,两者在细胞增殖、侵袭能力、离散能力等方面有差别。这种差异可能与c met有关。
Objective To screen B16M high and low metastatic strains to explore the heterogeneity and mechanism of B16M metastasis. Methods The cell lines with different metastatic potential were screened by B16M spontaneous lung metastasis model and their growth and metastasis in vivo were verified by experimental and spontaneous tumor metastasis models. Differences in proliferation, invasion and response to discrete factors in high and low metastatic B16M cell lines were studied by cell proliferation, 3 H TdR incorporation assay, flow cytometry and cell invasion assay. Western blotting was used to investigate the effect of c - met expression and activation level. Results In the three suspected high metastatic B16M cell lines, the weight of primary tumor formed by inoculation of H3B16M cells in the same line of C5 7BL / 6 mice was significantly higher than that of other cells Strain; The cell proliferation, invasiveness and ability to respond to discrete factors were significantly higher than other cell lines. c met expression and phosphorylation also increased. Conclusion The B16M high and low metastatic cell lines were screened successfully, both of which have differences in cell proliferation, invasion ability and discrete ability. This difference may be related to c met.