VEGF-C与COX-2反义寡核苷酸联合抑制乳腺癌细胞VEGF-C的表达  被引量:1

Inhibitory effect of combined transfection of antisense oligonucleotides of VEGF-C and COX-2 on the expression of VEGF-C in breast carcinoma cell line

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作  者:李昌秀[1] 曹友德[1] 黄源 

机构地区:[1]重庆医科大学病理学教研室,重庆400016 [2]重庆市开县人民医院神经外科,重庆405400

出  处:《肿瘤》2008年第2期117-120,共4页Tumor

摘  要:目的:探讨以阳离子脂质体(liposomes,LIP)为载体联合转染VEGF-C与COX-2的反义寡核苷酸(antisense oligonudeotides,ASODN)对乳腺癌细胞MDA-MB-231中VEGF-C表达的影响。方法:MTT法检测ASODN对细胞的毒性作用并确定最佳转染浓度,RT-PCR法检测联合转染VEGF-C与COX-2 ASODN后对乳腺癌细胞中VEGF-C mRNA表达的影响,免疫组织化学及激光共聚焦法检测联合转染后乳腺癌细胞VEGF-C蛋白的表达。结果:ASODN+LIP以浓度0.2mmol/L+2.5mL/L转染时对细胞无明显毒性作用;联合组对VEGF-C mRNA的抑制率为70.37%,单用VEGF-C及COX-2ASODN组分别为48.15%及42.21%,联合组对VEGF-C mRNA的抑制率显著高于单用ASODN组(P<0.01);联合组乳腺癌细胞中VEGF-C蛋白的表达明显低于单用VEGF-C及COX-2 ASODN组(P<0.01)。结论:联合转染VEGF-C与COX-2 ASODN可明显降低乳腺癌细胞MDA-MB-231中VEGF-C mRNA及蛋白水平。COX-2可能参与并诱导了乳腺癌细胞VEGF-C的表达。Objective:To investigate the effect of combined transfection of antisense oligonucleotides (ASODNs) of vascular endothelial growth factor-C (VEGF-C) and cyclooxygenase-2 (COX-2) by cationic liposome (LIP) on the expression of VEGF-C in breast carcinoma MDA-MB-231 cells. Methods:Cytotoxicity of ASODNs on MDA-MB-231 cells was measured by MTT assay and the optimal concentration for transfection was determined. The transcription of VEGF-C in breast carcinoma cells was detected by RT-PCR. The protein expression of VEGF-C was detected by immunohistochemical staining and confocal laser scanning microscopy in MDA-MB-231 cells after co-transfection of VEGF-C and COX-2 ASODN. Results:No cytotoxic effect was found when ASODNs/LIP was transfected at 0.2 mmol/L+2.5 mL/L. The inhibitory ratio on VEGF-C mRNA transcription was 70.37% in the combined transfection group, which was significantly higher than VEGF-C transfection group and COX-2 transfection group (48.15% and 42.21%, P〈0.01). The protein expression of VEGF-C was significantly lower in the combined transfection group than that in only VEGF-C or COX-2 transfection group (P〈0.01). Conclusion: Combined transfection of VEGF-C and COX-2 ASODN markedly decreases the expressions of VEGF-C at mRNA and protein levels in MDA-MB-231 cells. COX-2 maybe involved in the induction of VEGF-C expression in breast cancer.

关 键 词:乳腺肿瘤 血管内皮生长因子C 环氧化酶2 寡核苷酸类 反义 脂质体 

分 类 号:R737.9[医药卫生—肿瘤]

 

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