反义bcl-2硫代磷酸寡脱氧核苷酸对小细胞肺癌细胞株NCI-H446的抑制作用及诱导细胞凋亡的研究  

Antisense bcl-2 oligodeoxynucleotides inhibited the cell proliferation and induced apoptosis in a small-cell lung cancer cell line NCI-H446

作　　者：陈鑫基[1] 胡建达[1] 战榕[1] 吕联煌[1] 

机构地区：[1]福建医科大学附属协和医院

出　　处：《中国病理生理杂志》2003年第3期374-378,共5页Chinese Journal of Pathophysiology

摘　　要：目的 :观察反义bcl- 2硫代磷酸寡脱氧核苷酸 (AS -PS -ODN)对小细胞肺癌细胞株NCI-H44 6mRNA、蛋白以及增殖、活力和凋亡的影响。方法 :合成bcl- 2AS -PS -ODN作用于小细胞肺癌细胞株NCI-H44 6,半定量RT -PCR检测bcl- 2mRNA表达 ;免疫细胞化学染色和流式细胞仪检测Bcl- 2蛋白表达 ;通过克隆形成率、细胞计数、流式细胞仪DNA倍体分析、TUNEL等指标观察bcl- 2AS -PS -ODN对细胞增殖、活力和凋亡的影响。结果 :①bcl- 2AS -PS -ODN能特异性地降低NCI-H44 6细胞bcl- 2mRNA和Bcl- 2蛋白的表达。 1μmol/LAS -PS-ODN作用 2 4h后bcl- 2mRNA表达量下降 69 5 % ,48h后Bcl- 2蛋白下降 62 7%。②bcl- 2AS -PS -ODN能够抑制细胞增殖和活力 ,诱导细胞凋亡 ,1μmol/LAS -PS -ODN作用 2 4h后 ,细胞凋亡率约为 2 2 3 % - 3 2 7%。 结论 :bcl- 2AS -PS -ODN能够特异性降低bcl- 2mRNA、蛋白表达 ,抑制NCI-H44 6细胞的增殖和活力 。AIM: To observe the effect of antisense bcl-2 oligodeoxynucleotides(AS-PS-ODN) on bcl-2 mRNA and protein expression, cell proliferation,viability and apoptosis in a small-cell lung cancer cell line NCI-H446. METHODS: Semi-quantitative RT-PCR was performed to detect the bcl-2 mRNA expression, the Bcl-2 protein was determined by immunocytochemistry and flow cytometry analysis, and the effect of bcl-2 AS-PS-ODN on cell proliferation, viability and apoptosis were investigated by colony assay , cell count, DNA content analysis and TUNEL. RESULTS: ① 1 μmol/L bcl-2 AS-PS-ODN significantly down-regulated the expression of bcl-2 mRNA and protein. The inhibition rate of mRNA and protein were 69.5% and 62.7%, respectively. ② bcl-2 AS-PS-ODN decreased cell proliferation and viability , induced cell apoptosis.The apoptosis rate was 22.3%-32.7% in cells treated with 1μmol/L bcl-2 AS-PS-ODN. CONCLUSION: bcl-2 AS-PS-ODN down-regulated expression of bcl-2 mRNA and protein, inhibited cell proliferation and induced apoptosis in a small cell lung cancer cell line, NCI-H446.

关 键 词：硫代磷酸寡脱氧核苷酸 小细胞 肺癌细胞株 NCI-H446 BCL-2 细胞凋亡 

分 类 号：R734.2[医药卫生—肿瘤]