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目的:探讨脂质体介导的c-myc反义寡核苷酸对导入脆性组氨酸三联体(FHIT)基因的胃癌细胞增殖及凋亡的影响.方法:通过脂质体将重组FHIT基因PRC/ CMV质粒和空载体转染到人类胃癌细胞系MKN28,并分别转染c-myc反义寡核苷酸,RT- PCR和Westen blot法检测FHIT基因的转染,Western blot法检测细胞c-myc的表达,MTT法分析细胞增殖,AO/EB染色法和流式细胞分析技术检测细胞凋亡.结果:转染FHIT基因后,MKN28细胞检测到FHIT基因片段和FHIT蛋白,而未转染的细胞及转染空载体的细胞未检测到FHIT基因片段及FHIT蛋白.转染c-myc反义寡核苷酸后.对MKN28细胞c-myc的表达有明显的抑制作用,并呈明显的时间依赖性;c-myc asODN对FHIT~+ MKN28细胞抑制率(F=177.480,P<0.05),凋亡率(F=41.500,P<0.05)和凋亡比例明显高于FHIT MKN28细胞。结论:癌基因c-myc的表达抑制联合FHIT基因的表达可以发挥较强的抗肿瘤细胞作用,为多基因治疗肿瘤提供了理论基础.
Objective: To investigate the effect of liposome-mediated c-myc antisense oligonucleotide on the proliferation and apoptosis of gastric cancer cells induced by FHIT gene.Methods: The recombinant FHIT gene PRC / CMV plasmid and empty vector were transfected into human gastric cancer cell line MKN28 and transfected with c-myc antisense oligonucleotides respectively. Transfection of FHIT gene was detected by RT-PCR and Westen blot. Western blot was used to detect the expression of c -myc expression of, the MTT cell proliferation were analyzed, AO / EB staining and flow cytometry to detect apoptosis results: FHIT gene after transfection, cells MKN28 detected FHIT gene fragment and FHIT protein, without transfection FHIT gene fragment and FHIT protein were not detected in the cells transfected with empty vector.The transfected c-myc antisense oligonucleotides significantly inhibited the expression of c-myc in MKN28 cells, (F = 177.480, P <0.05). The apoptosis rate of FHIT ~ + MKN28 cells was significantly higher than that of FHIT MKN28 cells (F = 41.500, P <0.05). CONCLUSION: Inhibition of c-myc oncogene expression in combination with FHIT gene can exert a strong anti-tumor effect and provide a theoretical basis for multi-gene therapy of tumors.