反义c-myc寡核苷酸对转染FHIT基因胃癌细胞MKN28的影响

Effect of anti-sense c-myc oligodeoxynucleotide on MKN28 gastric cancer cells transfected with FHIT gene

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.

AIM： To explore the effect of anti-sense c-myc oligodeoxynucleotide on the proliferation and apoptosis of gastric cancer cells transfected with the fragile histindine triad （FHIT） gene. METHODS： The FHIT gene was transfected into human gastric cancer MKN28 cells through liposornes. The antis-sense c-rnyc oligodeoxynucleotide was transfected into the FHIT gene. FHIT gene transfection was detected by RTPCR and Western blot. Expression of c-myc was detected by Western blot. Proliferation of gastric cancer cells was determined by MTT. Cell apoptosis was detected by AO/EB staining and FCM. RESULTS： The expression of MKN28 cells transfected with the FHIT gene was observed. However, no FHIT gene segment and FHIT protein were found in the cells transfected with void vector. C-myc transfected with the FHIT gene exhibited significant inhibitory effects on the expression of c-myc protein in MKN28 cells and induced cell apoptosis in a time-dependent manner. However, the inhibition rate （F = 177.480, P 〈 0.05） （F = 41.500, P 〈 0.05）, apoptosis rate, and apoptosis portion of anti-sense c-myc oligodeoxynucleotide for C-myc were higher than those for FHIT^＋ MKN28 cells. CONCLUSION： The expression of c-myc and FHIT oncogenes plays an important role in suppressing the growth of tumor cells and may provide the theoretical basis for tumor gene therapy.