shRNA-CXCR4作用于CXCR4基因对乳腺癌血管生成潜力的影响

Effect of small interfering RNA targeting CXCR4 on breast cancer angiogenesis

目的研究shRNA-CXCR4作用于CXCR4基因对人乳腺癌细胞株诱导血管内皮细胞形成管腔能力的影响。方法通过质粒shRNA-CXCR4沉默CXCR4基因后,RT-PCR、Western-blot检测3株人乳腺癌细胞的CXCR4mRNA及其蛋白的表达;应用人乳腺癌细胞株与人脐静脉内皮细胞(HUVEC)双室联合培养技术,观察人乳腺癌细胞株通过shRNA-CXCR4作用于CXCR4基因后诱导HUVEC形成管腔能力的影响。结果质粒shRNA-CXCR4作用于3株人乳腺癌细胞MDA-MB-231、MDA-MB-435S、MCF-7后能明显抑制其CXCR4基因的mRNA及蛋白表达水平;对乳腺癌细胞诱导的人脐静脉血管内皮细胞HUVEC形成管腔样结构的能力有显著的抑制作用(P<0.05)。结论shRNA-CXCR4作用于CXCR4基因后能明显抑制人乳腺癌细胞诱导管腔形成能力。

Objective To study the effects of a small interfering RNA targeting CXCR-4 （shRNA-CXCR4） on angiogenesis of human breast cancer cells. Methods The expression ofCXCR4 mRNA and protein in 3 breast cancer cell lines with CXCR-4 silencing mediated by shRNA-CXCR4 was detected by RT-PCR and Western blotting, respectively. The morphological changes of human umbilical vein endothelial cells （HUVECs） were observed in co-culture with human breast cancer cells after CXCR4 gene silencing. Results CXCR4 mRNA and protein expressions decreased significantly in MCF-7, MDA-MB-231 and MDA-MB-435s breast cancer cells after the gene silencing （P〈0.05）. Gene silencing with shRNA-CXCR4 in human breast cancer cells significantly inhibited the ability of HUVECs to form tubular structures in the co-culture （P〈0.05）. Conclusion Gene silencing by shRNA-CXCR4 can obviously lower the angiogenesis-inducing ability of human breast cancer cells.