摘要
目的观察骨桥蛋白(OPN)基因沉默对卵巢癌细胞系HO-8910PM细胞的影响,为探索卵巢癌基因治疗提供理论依据。方法以脂质体法将已构建的骨桥蛋白特异性小干扰RNA真核表达载体(pSilencer3.1/OPN-shRNA)转染卵巢癌细胞HO-8910PM,48h后用Western blotting技术检测重组质粒组、空质粒组和空白对照组中卵巢癌细胞内骨桥蛋白的表达情况;采用四甲基偶氮唑蓝(MTT)法连续5天测定转染后细胞的增殖情况并绘制曲线图。结果 pSilencer3.1/OPN-shRNA转染卵巢癌细胞株HO-8910PM后48h骨桥蛋白表达下降,差异有统计学意义(同空质粒组、空白对照组相比P<0.05);空质粒组与空白对照组相比差异无统计学意义(P>0.05)。重组质粒组同空质粒组、空白对照组比较其增殖于第1天差异无统计学意义(P>0.05),第2～5天差异有统计学意义(P<0.05);空质粒组与空白对照组比较,第1～5天其增殖差异均无统计学意义(P>0.05)。结论构建的骨桥蛋白特异性小干扰RNA真核表达载体可以抑制卵巢癌细胞HO-8910PM中骨桥蛋白的表达。骨桥蛋白可能与卵巢癌细胞HO-8910PM的增殖抑制相关。
Objective To investigate the effect of OPN gene silence on ovarian cancer cell line HO-8910PM and to provide a theoretical basis for the therapeutics of ovarian cancer.Methods OPN specific siRNA eukaryotic expression vector (pSilencer3.1 /OPN-shRNA) was constructed and transfected into the ovarian cancer cell line HO-8910PM by liposome method.48 h later,Western blotting was employed to detect the expression of OPN protein in the ovarian cancer cells in the specific plasmid group,nonspecific plasmid group and control group,respectively.The cell proliferation was determined by MTT method and was recorded in the diagram for the consecutive five days as of OPN siRNA transfection into ovarian cancer cell line HO-8910PM.Results The expression of OPN protein decreased 48 h after OPN siRNA transfection into ovarian cancer cell line HO-8910PM.The difference between specific group and other groups was significant (P 0.05) .Compared with nonspecific and control groups,the cell proliferation in specific group for the 1st day had no statistical significance (P > 0.05) ,while the difference in cell proliferation was significant in 2nd-5th day (P 0.05) .Conclusion The constructed OPN specific siRNA eukaryotic expression vector,pSilencer3.1 /OPN-shRNA,could inhibit the expression of OPN in the ovarian cancer cell line HO-8910PM.OPN may be correlated with the proliferation inhibition of ovarian cancer cell HO-8910PM.
出处
《徐州医学院学报》
CAS
2010年第8期504-506,共3页
Acta Academiae Medicinae Xuzhou
