B16M高低转移株的筛选及其生物学异质性的分析

Screening out high and low metastases cell line of B16 melanoma and the analysis of their biological heterogeneity

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 out high and low metastases B16 Melanoma cell lines for investigating the molecular mechanisms of B16 Melanoma metastases.Methods High and low metastases B16 Melanoma cell lines were screened by tumor spontaneous lung metastases model and confirmed by experimental or spontaneous lung metastases model. The biological abilities of these cell lines were analyzed by cellular biological tests, such as cell proliferation experiment, 3H-TdR, FACS, tumor invasion and scatter test. Expression and phosphorylation of c-met in the cell lines were detected by western blot. Results Four B16M cell lines (named as L H1 H2 H3) were screened ont and H3 cell line displayed a high metastases or growth ability in tumor metastases test in C 57 BL/6 mouse in vivo. The biological ability of the cell in proliferation, DNA synthesis and invasion in vitro of H3 B 16 M was super to the other cell lines. Furthermore, the higher level of c-met expression and phosphorylation was found in H3 B16 Melanoma cell lines. Conclusion We screened out low metastases B16 Melanoma cell lines and high metastases B16 Melanoma cell lines. Their significant differences in the biological abilities may be related to c-met expression and phosphorylation.