骨桥蛋白反义寡核苷酸对微缺氧下人结肠癌细胞HT-29增殖活性及侵袭力的影响

目的 观察靶向骨桥蛋白(OPN)的反义寡核苷酸(ASODN)对微缺氧下HT-29细胞增殖活性、侵袭转移潜能等的影响,探讨OPN与微缺氧诱导的肿瘤恶性表型的关系.方法 合成ASODN,以Lipofectamine为载体,将其转染入微缺氧下高表达OPN的HT-29细胞.用RT-PCR和Western blot法分别检测转染细胞微缺氧下OPN mRNA和蛋白的表达,MTT检测转染的HT-29细胞微缺氧下的增殖活性,MTT比色试验测定HT-29细胞的异质性黏附能力,明胶酶谱分析检测分泌的MMP-2/9活性变化.结果 微缺氧诱导的OPN mRNA和蛋白表达被靶向OPN的ASODN特异性地抑制,表达量分别是对照组的31.5%和35.4%.ASODN组HT-29细胞的吸光度值(A570)明显低于各对照组(P<0.01),生长抑制率(IR)达(41.6±1.2)%.ASODN组HT-29细胞在各时限点的黏附率均明显降低,明胶酶活性下降71%,与各对照组比较差异有统计学意义(P<0.01).结论 靶向OPN的ASODN明显抑制微缺氧诱导的OPN mRNA和蛋白表达,显著拮抗微缺氧诱导的细胞增殖、异质性黏附,并显著下调微缺氧诱导的细胞分泌MMP2/9的活性.OPN在微缺氧促进肿瘤向恶性表型转化中起着重要作用.

Effects of antisense oligonucleotide targeting osteopontin on proliferation and invasion of colon carcinoma cell lines HT-29 in vitro under moderate hypoxia

Objective To study the effects of antisense oligonucleotide(ASODN) targeting osteopontin on proliferation and the invasiveness of HT-29 cells under moderate hypoxia.To investigate correlation between osteopontin and malignant phenotype induced by moderate hypoxia.Methods ASODN targeting osteopontin were synthesized with a phosphorothioate backbone.Mediating by lipofectamine,ASODN was transfected into HT-29 cells with high expression of osteopontin induced by moderate hypoxia.The osteopontin mRNA and protein...