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目的探讨REV3基因低表达联合抗癌药物对人结肠癌细胞(SW-480)增殖和凋亡的影响。方法利用RNAi技术降低REV3基因在SW-480细胞中的表达,以实时荧光定量PCR和免疫细胞化学检测REV3表达量的降低情况,选择低表达效率具有统计学意义的细胞作为实验组细胞,再用抗癌药物作用48h后运用MTT实验和流式细胞仪,对实验组和对照组细胞进行细胞增殖和凋亡变化情况的检测。结果与空白对照组相比,REV3基因低表达或抗癌药物(顺铂或槐定碱)单独作用均能促进SW-480细胞凋亡并抑制其增殖(P<0.01);实验组内,与药物空白组相比,REV3基因低表达的同时再联合一种抗癌药物(顺铂或槐定碱),对SW-480细胞凋亡和增殖的作用较单一因素更加显著;若将三者联合则比以上任一两种联合作用效果更为显著,以上结果均具有统计学意义(P<0.05)。结论 REV3基因低表达、顺铂、槐定碱或这三因素两两联合均可促进结肠癌SW-480细胞的凋亡并抑制其增殖,若三者同时联合作用效果更加显著,提示“基因+药物”联合抑癌具有协同增效的作用。
Objective To investigate the effect of low expression of REV3 gene combined with anticancer drugs on proliferation and apoptosis of human colon cancer cells (SW-480). Methods RNAi was used to reduce the expression of REV3 gene in SW-480 cells. Real-time quantitative PCR and immunocytochemistry were used to detect the decrease of REV3 expression. The cells with low expression efficiency were selected as the experimental group cells, The effect of anticancer drugs 48h MTT assay and flow cytometry, the experimental group and control group cell proliferation and apoptosis detection. Results Compared with the blank control group, the low expression of REV3 gene or the anti-cancer drug alone (cisplatin or sophoridine) could promote the apoptosis and inhibit the proliferation of SW-480 cells (P <0.01) Compared with the blank control group, the combination of an anti-cancer drug (cisplatin or sophoridine) and the low expression of REV3 gene in SW-480 cells was more significant than single factor. Compared with either of the above two joint effects more significant, the above results were statistically significant (P <0.05). Conclusion Low expression of REV3 gene, cisplatin, sophoridine or combination of these three factors can promote the apoptosis of colon cancer SW-480 cells and inhibit its proliferation. If the combination of these three factors is more effective, it suggests that the gene “ + Drugs ”Joint suppression of cancer has synergistic effect.