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目的探讨Bcl-XL反义寡核苷酸转染后,对食管癌细胞增殖及裸鼠体内人食管癌移植瘤生长的影响。方法采用阳离子脂质体介导的反义寡核苷酸转染、逆转录聚合酶链反应(RT-PCR)、Western蛋白印迹杂交、四甲基偶氮唑盐光吸收法(MTT法)、流式细胞术及凋亡的原位末端标记(TUNEL)检测等方法观察了Bcl-XL反义寡核苷酸转染对食管癌细胞增殖、凋亡的影响及裸鼠体内人食管癌移植瘤生长的影响。结果MTT法检测显示,Bcl-XL反义寡核苷酸可显著抑制食管癌细胞的增殖(P<0.05);对Bcl-XLmRNA表达抑制率为57.3%,可显著下调Bcl-XL的蛋白表达;应用流式细胞术和TUNEL检测技术,反义寡核苷酸组的凋亡率分别为(31.1±5.8)%和35.0%,与相关对照组比较差异有统计学意义(P<0.01)。反义寡核苷酸组裸鼠体内人食管癌移植瘤的生长受到显著抑制(P<0.05)。Bcl-XLmRNA及蛋白表达亦受到明显抑制,并且移植瘤组织中凋亡细胞明显增加。结论Bcl-XL反义寡核苷酸可有效抑制食管癌细胞增殖及裸鼠体内人食管癌移植瘤的生长。通过反义寡核苷酸下调Bcl-XL的表达,达到治疗食管癌的目的,为食管癌的基因治疗提供实验依据。
Objective To investigate the effect of Bcl-XL antisense oligonucleotide transfection on the proliferation of esophageal cancer cells and the growth of human esophageal carcinoma in nude mice. Methods Liposome-mediated antisense oligonucleotide transfection, reverse transcription-polymerase chain reaction (RT-PCR), Western blotting, MTT assay, Flow cytometry and apoptosis in situ TUNEL detection of Bcl-XL antisense oligonucleotide transfection on esophageal cancer cell proliferation and apoptosis and in vivo human esophageal cancer xenografts The impact of growth. Results The results of MTT assay showed that Bcl-XL antisense oligonucleotide significantly inhibited the proliferation of esophageal cancer cells (P <0.05), and inhibited the expression of Bcl-XL mRNA by 57.3% The apoptosis rate of antisense oligodeoxynucleotide group was (31.1 ± 5.8)% and 35.0% respectively by flow cytometry and TUNEL assay, which showed significant difference compared with the control group (P <0.01). The growth of human esophageal carcinoma transplanted with antisense oligodeoxynucleotide in nude mice was significantly inhibited (P <0.05). Bcl-XL mRNA and protein expression were also significantly inhibited, and the apoptotic cells in xenograft tumors significantly increased. Conclusion Bcl-XL antisense oligonucleotide can effectively inhibit the proliferation of esophageal cancer cells and the growth of human esophageal carcinoma in nude mice. Through the down-regulation of Bcl-XL expression by antisense oligonucleotide to achieve the purpose of treatment of esophageal cancer, to provide experimental evidence for the gene therapy of esophageal cancer.