【摘要】 目的:探讨转染67kDa层粘连蛋白受体(67kDa laminin receptor,67kDa LN-R)反义寡核苷酸(antisense oligonucleotide,ASODN)后卵巢癌细胞HRA的生物学行为变化。方法:合成67 kDa LN-R反义及正义寡核苷酸片段(sense oligonucleotide SODN),设定反义组、正义组及对照组并分别转染卵巢癌细胞HRA;采用形态学观察、MTT、流式检测各组细胞生长情况;免疫组化检测各组细胞67 kDa LN-R蛋白表达;Transwell小室法检测67 kDa LN-R反义寡核苷酸对卵巢癌细胞体外侵袭转移能力的影响。结果:转染24￣48 h内细胞增殖受到不同程度抑制,48 h后细胞生长状态有所恢复。0.02、0.04、0.08μg/μlASODN处理24 h后细胞凋亡率分别为19.38%、21.62%、30.35%,差异有统计学意义(P<0.05)。转染反义寡核苷酸组67 kDa LN-R的表达逐渐减少,且下调作用与ASODN浓度呈剂量依赖性,与正义组及对照组相比差异有显著性(P<0.05);转染后HRA穿透人工基底膜的能力显著降低并呈现剂量依赖性(P<0.05)。结论:67 kDa LN-R ASODN可下调卵巢癌细胞HRA67 kDa LN-R表达并降低其体外侵袭转移能力。更多还原

【Abstract】 Objective: To investigate the effects of 67kDa laminin receptor(67kDa LN-R) antisense oligonucleotide(ASODN) on biological behavior of ovarian carcinoma cell HRA in vitro.Methods: The 67 kDa LN-R ASODN and its control,sense oligonucleotide(SODN) were synthesized and transfected into the target cells.The survival situation and the induced-apoptosis effect were detected by MTT assay and FCM.Immunocytochemistry method was used to evaluate the 67 kDa LN-R protein expression levels to identify the efficiency of ASODN.The invasiveness of transfected cells was measured quantitatively by Matrigel invasion assays(Transwell chamber).Results: The ovarian carcinoma cells treated with ODNs for 24 to 48 hours were inhibited and partially regained the normal situation of growth after changing the medium.Transfected with 0.02,0.04 and 0.08 μg/μl ASODN for 24 hours,19.38%,21.62% and 30.35%of the HRA cell apoptosis were observed(P<0.05).The level of 67 kDa LN-R protein expression and invasiveness of HRA cells treated with ASODN of different final concentration were significantly decreased compared with the results from those transfected with SODN and the controls(P<0.05).Conclusion: Down-regulating the level of 67 kDa laminin receptor,67 kDa LN-R ASODN has a significant inhibitory effect on the invasiveness of human ovarian carcinoma cell line HRA in a dose-dependent manner.更多还原