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目的研究RNA干扰对透明质酸酶基因HYAL1的表达以及对人乳腺癌细胞侵袭力的影响。方法体外化学合成HYAL1序列特异性双链RNA(dsRNA),在脂质体(SiPORT Lipid)的介导下转染入人乳腺癌细胞株MDA-MB-231、MDA-MB-453S、ZR-75和ZR-75-30。荧光共聚焦显微镜下观察转染效率,RT-PCR分析HYAL1 mRNA的表达,体外实验测定肿瘤细胞穿透matrigel的能力。结果4种乳腺癌细胞株的HYAL1基因的表达均被HYAL1-siRNA有效封闭,HYAL1 mRNA相对水平明显降低(P<0．05)。转染48 h后4种人乳腺癌细胞的侵袭力均明显降低(P<0．01)。结论HYAL1-siRNA能有效抑制人乳腺癌细胞株HYAL1基因的表达,降低人乳腺癌细胞的侵袭力。
Objective To study the effect of RNA interference on the expression of hyaluronidase gene HYAL1 and its invasiveness in human breast cancer cells. Methods HYAL1 sequence specific double stranded RNA (dsRNA) was chemically synthesized in vitro and transfected into human breast cancer cell lines MDA-MB-231, MDA-MB-453S and ZR-75 under the guidance of SiPORT Lipid And ZR-75-30. Transfection efficiency was observed under fluorescence confocal microscope, the expression of HYAL1 mRNA was analyzed by RT-PCR, and the ability of tumor cells to penetrate matrigel was determined in vitro. Results The expression of HYAL1 gene in all four breast cancer cell lines was effectively blocked by HYAL1-siRNA, and the relative level of HYAL1 mRNA was significantly decreased (P <0.05). The invasiveness of the four human breast cancer cells was significantly decreased 48 h after transfection (P <0.01). Conclusion HYAL1-siRNA can effectively inhibit the expression of HYAL1 gene in human breast cancer cell line and reduce the invasiveness of human breast cancer cells.