FAK反义寡核苷酸对卵巢癌细胞凋亡、黏附和侵袭性的影响.docVIP

FAK反义寡核苷酸对卵巢癌细胞凋亡、黏附和侵袭性的影响 【摘要】目的　观察黏着斑激酶的反义寡核苷酸对卵巢腺癌细胞株OVCAR-3凋亡、黏附和侵袭性的影响。方法　实验分4组，反义寡核苷酸组、正义寡核苷酸组、脂质体组、空白对照组。用脂质体介导的方法，将黏着斑激酶的反义寡核苷酸转入OVCAR-3卵巢癌细胞株，应用流式细胞仪Annexin V/PI双染色法检测细胞的凋亡率，应用基质胶细胞黏附实验和Boyden小室法检测卵巢癌细胞的体外黏附和侵袭能力。结果　ASODN组卵巢癌细胞的凋亡率与对照组比较差异有统计学意义(P0.05);ASODN组癌细胞与基质胶的粘附率与对照组比较差异有统计学意义(P0.05);ASODN组癌细胞穿透Matrigel胶的细胞数目与对照组比较差异具有统计学意义(P0.05)。结论　抑制黏着斑激酶的表达能增加卵巢癌细胞株OVCAR-3的凋亡，并降低其黏附和侵袭转移能力。 【关键词】 黏着斑激酶;卵巢癌;侵袭;凋亡;粘附 【Abstract】　Objective　To investigate the inhibitory effects of antisense oligodeoxynucleotides (ASODN) of focal adhesion kinase (FAK) on the apoptosis，adhesion and invasion of ovarian carcinoma cell line-OVCAR-3.Methods　The ASODN was transfected into OVCAR-3 cells with lipofectamine TM 2000 in the form of liposome-ODN complexes.The cell apoptosis was observed by flow cytometry with Annexin-V/PI dual staining.The cell attachment assay was carried out in a culture well pre-coated with matrigel.The adhesion and invasion of ovarian cancer in vitro were detected by matrigel cell adhesion testing and Boyden chamber method.Results　The apoptosis rate of OVCAR-3 cells treated with ASODN was increased significantly，as compared with that in control group (P0.05).The adhesion rate of OVCAR-3 cells in matrigel treated with ASODN was decreased significantly，as compared with that in control group (P0.05).The number of OVCAR-3 cells that penetrated the matrigel was decreased significantly，as compared with that in control group(P0.05).Conclusion　The inhibition of FAK expression can increase the apoptosis of ovary carcinoma cell line-OVCAR-3，and can decrease its adhesion and invasion ability. 【Key words】　FAK;ovarian carcinoma;invasion;apoptosis;adhesion 卵巢癌是常见的妇科恶性肿瘤之一，主要的死亡原因是转移，但是目前发生转移的机制尚不完全明了。黏着斑激酶 (focal adhesion kinase，FAK)是一种新近发现的酶类，临床研究显示其在多种人类肿瘤中均有表达，与肿瘤的生长、侵袭与转移特性关系密切[1-4]。我们尝试以FAK的反义寡核苷酸(antisense oligonucleotide，ASODN)体外转染卵巢癌细胞系OVCAR3，观察其对癌细胞凋亡、黏附和侵袭能力的影响。 1　材料与方法 1.1　实验材料 脂质体lipofectin 2000购自美国Invitrogen公司。Transwell小室购于美国Coming Costar公司。Matrigel胶购于BD Bios