Bcl-2反义寡核苷酸对人小细胞肺癌细胞NCI-H69化疗敏感性的检测

Effect of Bcl-2 Antisense Oligodexynucleotides on Chemotherapy Efficacy of Vp-16 on Human Small Cell Lung Cancer Cell Line NCI-H69

目的:研究Bcl-2反义寡核苷酸对以NCI-H69为标记的人小细胞肺癌细胞化疗药物敏感性的促进作用。方法:将NCI-H69细胞培养传代,细胞共分4组,反义寡核苷酸组、正义寡核苷酸组、无义寡核苷酸组和空白对照组,通过脂质体转染将不同寡核苷酸导入细胞中,Western-Blot法检测Bcl-2蛋白的表达。化疗药物依托泊甙(Vp-16)分别以不同浓度加入4组细胞。MTT法测定细胞存活分数。结果:4组细胞中加入Vp-16后,反义寡核苷酸组细胞在Vp-16浓度0.25、0.5、1.0、2.0、4.0μg/ml的凋亡率明显高于空白对照组,统计结果差异具有显著性(P<0.05),正义寡核苷酸组和无义寡核苷酸组与空白对照组相比则无统计学差异(P>0.05)。转染反义寡核苷酸组在不加入Vp-16和加入Vp-16不同剂量后细胞的存活分数均明显低于空白组,且有统计学差异(P<0.01),而转染正义链和无义链组与空白组比较则无统计学差异(P>0.05)。结论:Bcl-2反义寡核苷酸能有效封闭Bcl-2基因的表达,增强小细胞肺癌细胞NCI-H69对化疗药物的敏感性。

Objective To study the effect of bcl - 2 antisense oligodexynucleotides on chemotherapy efficacy of Vp - 16 on human small cell lung cancer cell line NCI - H69. Methods Cultured NCI - H69 cells were devided into 4 groups： bcl - 2 antisense oligodexynucleotides （ASODN） added, sense oligodexynucleotides （SODN） added, nonsense oligodexynucleotides （NSODN） added and control （no nucleotides added）, the oligodexynucleotides were transfected into the cultured cells with oligofectamine. The cellular expression of Bcl - 2 protein 72h later was examined with Western - Blot. The four different groups of cuhured tumor cells were treated with etopside（ Vp - 16） at different concentrations （0, 0.25, 0.5, 1.0, 2.0 and 4.0 μg/ml） for 48hr then the cell survival fraction was assessed with MTT test. Results The apoptotic rate of ceils in the ASODN group was significantly higher than that of the control group, also, the survival fraction of cells in ASODN group was significantly lower than that of the control group. The Bcl - 2 protein expression in ASODN group was significantly lower than that in the control group, but no inhibition was observed in SODN and NSODN groups. Conclusion The bc1-2 ASODN could enhance the sensitivity to chemotherapy with Vp - 16 in small cell lung cancer cell line NCI- H69 by effectively blocking bcl- 2 gene expression.