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目的研究Bcl-2反义寡核苷酸是否能增强小细胞肺癌细胞NCI-H69对化疗药物的敏感性。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,通过脂质体介导的方法将不同寡核苷酸导入细胞中,Western-Blot法检测Bcl-2蛋白的表达。不同浓度的顺铂与阿霉素作用于4组转染后的细胞,MTT法测定细胞存活分数。结果4组细胞中,与空白对照组相比较,反义寡核苷酸组的Bcl-2蛋白表达明显受到抑制,而正义寡核苷酸组和无义寡核苷酸组细胞的Bcl-2蛋白表达则与空白对照组差别不明显。细胞分别同顺铂与阿霉素作用后,反义寡核苷酸组细胞在各个浓度的存活分数均低于空白组,其差异在统计学上有显著性意义(P<0.01),而转染正义链和无义链组与空白组比较其差异在统计学上无显著性意义。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对化疗药物的敏感性。
Objective To investigate whether Bcl-2 antisense oligonucleotides enhance the chemosensitivity of small cell lung cancer cells NCI-H69. Methods NCI-H69 cells were cultured and passaged. The cells were divided into 4 groups, antisense oligonucleotide group, sense oligonucleotide group, non-sense oligonucleotide group and blank control group. Different oligonucleotides were introduced into the cells, and the expression of Bcl-2 protein was detected by Western-Blot. Different concentrations of cisplatin and doxorubicin act on the four groups of transfected cells, MTT assay cell viability score. Results Compared with the blank control group, the expression of Bcl-2 protein in the antisense oligonucleotide group was significantly inhibited in the 4 groups of cells, whereas the expression of Bcl-2 in the cells of the sense oligonucleotide group and the non-sense oligonucleotide group was significantly decreased The protein expression was not significantly different from the blank control group. After cells were treated with doxorubicin and cisplatin respectively, the surviving fraction of antisense oligodeoxynucleotides in each concentration group was lower than that in blank group, and the difference was statistically significant (P <0.01) The difference between the stained sense and nonsense groups was statistically insignificant compared with the blank group. Conclusion Bcl-2 antisense oligonucleotides can effectively block the expression of Bcl-2 gene and enhance the sensitivity of small cell lung cancer cells to chemotherapeutic drugs.