非分泌型CXCL16在乳腺癌细胞系中的表达及对其生物学特性的影响

Expression of Non-secretory CXCL16 and Its Impact on Biological Characteristics in Breast Cancer Cell Lines

目的通过检测非分泌型CXCL16在不同侵袭性乳腺癌细胞系(SK-BR-3、MCF-7、MDA-MB-231、MDA-MB-435S)及正常乳腺上皮细胞MCF-10A的表达,探讨非分泌型CXCL16表达对乳腺癌细胞生物学特性的影响。方法采用RT-PCR检测CXCL16mRNA在MCF-10A、SK-BR-3、MCF-7、MDA-MB-231、MDA-MB-435S细胞系的表达,将pEGFP-CXCL16真核表达质粒转染到低表达CXCL16的MDA-MB-231细胞,通过实时荧光定量PCR和Western blot验证CXCL16过表达,采用体外迁移、侵袭实验及甲基四唑蓝(MTT)法分别检测过表达CXCL16对MDA-MB-231细胞迁移、侵袭和增殖活性的影响。结果 CXCL16mRNA在高侵袭性的MDA-MB-231细胞低表达,在低侵袭性的MCF-7细胞高表达,过表达CXCL16使得MDA-MB-231细胞的迁移、侵袭减弱,而增殖未受到影响。结论非分泌型CXCL16表达与乳腺癌细胞系的侵袭性相关,其表达可以抑制乳腺癌细胞的体外迁移、侵袭。

Objective To detect the expression of non-secretory CXCL16 and its impact on malignant biological behaviors in breast cancer cell lines. Methods RT-PCR was carried out to examine the expression of CXCL16 mRNA in breast cancer cell lines with different aggressiveness SK-BR-3, MCF-7, MDA-MB-231, MDA- MB-435S and human mammary normal epithelial celll MCF-10A. The eukaryotic expression plasmid of CXCL16 was transfected into MDA-MB-231 ceils and overexpression of CXCL16 was confirmed by Real time PCR and Western blot. Boyden Chamber assay was used to determine cell migration and invasion, while MTT assay was performed to determine cell proliferation. Results Among four breast cancer cell lines, CXCL16 mRNA was highly expressed in MCF-7, lowly expressed in MDA-MB-231, while MCF-IOA faintly expressed CXCL16. Overexpression of CXCL16 led to a decrease in cell migration, invasion but not proliferation in MDA-MB-231 cells. Conclusion Expression of non-secretory CXCL16 is associated with aggressiveness of breast cancer cell lines, and CXCL16 expression inhibits cell migration and invasion in vitro.