bcl-2反义寡核苷酸促进乳腺癌MCF-7细胞凋亡的研究

Apoptosis increased by bcl2 antisense ODN in MCF7 cell line

目的观察ｂｃｌ２反义寡核苷酸（ｏｌｉｇｏｎｕｃｌｅｏｔｉｄｅｓ，ＯＤＮ）对ＭＣＦ７细胞内ｂｃｌ２表达及细胞凋亡的影响。方法电转染法将合成的针对ｂｃｌ２ｍＲＮＡ的反义ＯＤＮ导入ＭＣＦ７乳腺癌细胞，用荧光显微镜、电镜、免疫组化法及流式细胞仪检测反义ＯＤＮ对乳腺癌细胞凋亡、ｂｃｌ２表达及细胞周期的影响。结果经ｂｃｌ２反义ＯＤＮ作用后，ＭＣＦ７细胞ｂｃｌ２基因表达降低、凋亡细胞数增多，作用前后差异有显著意义（Ｐ＜００５）；Ｇ０～Ｇ１期细胞减少，而Ｇ２～Ｍ期、Ｓ期细胞增多。结论ｂｃｌ２反义ＯＤＮ经电转染法导入乳腺癌细胞后，可以获得较好的瞬时表达，降低ｂｃｌ２功能，促进细胞凋亡。在肿瘤研究中，ｂｃｌ２可作为一个有用的靶基因。

bjective To study the effects of bcl2 antisense oligonucleotides(ODN) on the expression of bcl2 and apoptosis in MCF7 cell line. Methods Artificially synthesized bcl2 specific antisense OND was introduced into MCF7 cells by gene pulser electroprotocol. Fluorescence microscope, electron microscope, immunocytochemical staining assay, and FCM were used to detect the effects on cell apoptosis, expressional level of bcl2 protein and cell cycle after transfecting bcl2 antisense ODN. Results The number of apoptotic cells was increased and expression of bcl2 protein was declined (P<005). FCM showed that the number of G0G1 phase cells was decreased and the number of G2M, S phase cells were increased. Conclusions The effective transient expression of bcl2 antisense ODN can be obtained by gene pulser electroprotocol in MCF7 cell, the function of bcl2 gene was declined and the number of apoptotic cells was increased after transfecting bcl2 antisense ODN. bcl2 may be used as an useful target in tumor research.