摘要
目的研究凋亡抑制基因ｂｃ１－２的反义寡脱氧核苷酸（ＯＤＮ）对顺铂诱导肺癌细胞凋亡的促进作用。方法选择表达 ｂｅ１－２的肺癌细胞培养传代，分成 ７组：ＯＤＮ组、无意序列组、ＯＤＮ＋顺铂组、无意序列＋顺铂组、顺铂组、脂质体组和空白对照组。以脂质体与 ｂｃ１－２ＯＤＮ混合导入培养的相应组肺癌细胞中，培养６ ｈ后，相应孔加入抗癌药顺铂，再培养 １６ ｈ终止细胞培养并做细胞涂片，每组重复６个孔。用免疫细胞化学检测ｂｅ１－２的表达。采用末端脱氧核苷转移酶（ＴｄＴ）介导的脱氧尿苷三磷酸（ｄＵＴＰ）末端标记法（ＴＵＮＥＬ）原位观察癌细胞凋亡，并计算平均凋亡指数（ＡＩ）。结果经ＯＤＮ处理的癌细胞 ｂｃ１－２表达明显减弱，抑制有效率为 ６０．８％。凋亡的癌细胞胞核染成棕褐色。 ＯＤＮ＋顺铂组ＡＩ为１６．４±１．７，ＯＤＮ组ＡＩ为５．９±０．２，顺铂组ＡＩ为４．１±０．８，无意序列组ＡＩ为３．３±０．７，无意序列＋顺铂组ＡＩ为７．６±１．１，脂质体组ＡＩ为５．１±０．９，空白对照组ＡＩ为３．６±０．６。ＯＤＮ＋顺铂组ＡＩ明显高于其他各实验组（Ｐ＜０．０１）。ＯＤＮ组与顺铂组的ＡＩ之和明显小于ＯＤＮ＋顺铂组（Ｐ＜０．０１）。
Objective To study the effect of antisense oligodeoxyribonucleotide (ODN) to apoptotic suppress gene bc1-2 on apoptosis of lung carcinoma cells induced by cisplatin.Methods The lung carcinoma cells expressing bc1-2 were chosen to participate in this experiment. Cultured cells were divided into 7 groups: ODN, nonsense, ODN + cisplatin, nonsense + ciplatin, cisplatin, lipofectin and control. Bc1-2 antisense or nonsense mixed wiht lipofectin was added into above corresponding cultured cells After cultured for 6 hours, cisplatin was added into corresponding groups. The cells were cultured again for 16 hours. And then, the cells were smeared on slides. Apoptotic cells were labeled with TaT-mediated dUTP nick end labeling (TUNEL) method on cell smears. Apoptotic index (AI) was counted to show the percentage of apoptotic cancer cells. The immunocytochemistry was used to detect the expression of bc1-2 in carcinoma cells.Results The bc1-2 expression of cancer cell in ODN group was significantly decreased compared to the control and nonsense groups. The AI of ODN + cisplatin group was 16.4± 1.7, cisplatin group 4.1 ±0.8, antisense group 5.9 ± 0.2, nonsense group 3.3 ± 0.7, nonsense + cisplatin 7.6± 1.1,lipofectin 5.1 ± 0.9, control group 3.6 ± 0.6. The AI of antisense + cisplatin group was significantly higherr than that of other groups.Conclusion Antisense oligodeoxyribonucleotide to bc1-2 can inhibit significantly the expression of bc1-2 of lung cancer cells and increase apoptosis of cancer cells induced by cisplatin.
出处
《中华结核和呼吸杂志》
CAS
CSCD
北大核心
2000年第12期722-724,I030,共4页
Chinese Journal of Tuberculosis and Respiratory Diseases
基金
卫生部科研基金资助项目
广州市科委科研基金(96-Z-58-2)
关键词
BCL-2
反义寡核苷酸
顺铂
肺癌
调亡
bc1-2
Antisense oligodeoxyribonucleotide
Cisplatin
Lung cancer
apoptosis
