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目的研究抗增殖蛋白家族成员TOB1对乳腺癌细胞MDA-MB-231体外侵袭与转移的生物学作用与机制。方法采用脂质体介导的质粒转染和G418筛选培养法获得稳定高表达TOB1的高转移性乳腺癌细胞系MDA-MB-231。通过人工基底膜侵袭实验和划痕实验检测TOB1过表达对MDA-MB-231细胞体外侵袭能力和转移能力的影响。采用super array基因芯片法检测TOB1引起的肿瘤转移相关基因表达的变化。结果与对照组相比,外源性TOB1高水平表达使MDA-MB-231的体外侵袭能力显著降低(P<0.05),同时抑制MDA-MB-231的体外迁移能力;TOB1表达水平的增加引起MDA-MB-231细胞中肿瘤转移抑制基因BRMS1表达的明显增加,同时显著抑制肿瘤转移相关基因HSP90、FXYD5、RHOC、S100A4等的表达(均P<0.05)。结论 TOB1过表达抑制乳腺癌细胞MDA-MB-231的体外侵袭、转移,涉及的机制可能与TOB1对多种重要的肿瘤转移相关基因的表达调控有关。
Objective To study the biological effects and mechanisms of TOB1, an antiproliferative protein family member, on the invasion and metastasis of breast cancer cell MDA-MB-231 in vitro. Methods Liposome-mediated plasmid transfection and G418 screening culture method were used to obtain high-metastatic breast cancer cell line MDA-MB-231 stably expressing TOB1. The effects of TOB1 overexpression on the invasion and metastasis of MDA-MB-231 cells in vitro were detected by artificial basement membrane invasion assay and scratch assay. The super array microarray was used to detect the gene expression of tumor metastasis caused by TOB1. Results Compared with the control group, high levels of exogenous TOB1 significantly reduced the invasiveness of MDA-MB-231 in vitro (P <0.05) and inhibited the migration of MDA-MB-231 in vitro. The increased expression of TOB1 The expression of BRMS1 in MDA-MB-231 cells was significantly increased, and the expressions of HSP90, FXYD5, RHOC and S100A4 were also significantly inhibited (all P <0.05). Conclusions TOB1 overexpression inhibits the invasion and metastasis of breast cancer cell MDA-MB-231 in vitro. The possible mechanism is that TOB1 may be involved in the regulation of many important tumor metastasis-related genes.