沉默HIF-1α基因逆转结肠癌MDR的研究

Research on silencing HIF-1α gene reversing MDR in colon cancer

目的探讨沉默HIF-1α基因对结肠癌多药耐药(MDR)逆转的影响。方法建立MDR1、HIF-1α的慢病毒和亲本HT-29细胞多细胞球的体系,低氧引导形成耐药的模型,分为Neg-miR感染组(A组)、亲本HT-29 MCS未处理(常氧)组(B组)、MDR1 miR感染组(C组)、亲本HT-29 MCS未处理(低氧)组(D组)、HIF-1αmiR感染组(E组),检测耐药扭转情况和肿瘤细胞死亡情况。结果 MDR1与HIF-1α的两套miR的重组慢病毒的干扰体系可以跟随细胞进行传代,能够显著对靶基因蛋白表达产生抑制作用;各组细胞周期比较,D组、E组和C组、B组对比差异均有统计学意义(P<0.05),E组与B组对比差异有统计学意义(P<0.05),C组与E组对比差异有统计学意义(P<0.05);各组敏感性和细胞凋亡率比较,D组(VCR、ADR、5-Fu)、C组(5-Fu)、E组(5-Fu)与B组对比差异有统计学意义(P<0.05),E组(VCR、ADR、5-Fu)、C组(ADR、VCR)与D组对比差异有统计学意义(P<0.05),C组(5-Fu)与E组对比差异有统计学意义(P<0.05)。结论沉默HIF-1α能够显著逆转HT-29MCS细胞对5-Fu、VCR和ADR的耐药性。

Objective To explore the effect of silencing HIF-1αgene for reversing MDR of colon cancer.Methods The slow virus of MDR1,HIF-1αand the multicellular spheroids system of the parental HT-29 cell were established.The drug resistance model was formed by the low-oxygen guide.They were divided into five groups,including Neg-miR infection group(group A),parent N group(group B),MDR1-miR infection group(group C),parent H group(group D)and HIF-1αmiR infection group(group E).Then the drug resistance reversion and tumor cell death were detected.Results The interference system of two sets of miR recombinant lentivirus with MDR1 and HIF-1 could be passaged by following cells,could significantly produce the inhibiting effect on the expression of the target gene protein(P<0.05);in the cell cycle:there was statistically significant difference between the group D and group E and between group C and group B(P<0.05).There was statistically significant difference between the group E and group B(P<0.05).There was statistically significant difference between the group C and group E(P<0.05).The sensitivity and apoptosis rate in each group:there was statistically significant difference among group D(VCR,ADR,5-Fu),group C(5-Fu),group E(5-Fu)and group B(P<0.05).There was statistically significant difference between group E(VCR,ADR,5-Fu)and group C(ADR,VCR)with group D(P<0.05).There was statistically significant difference between the group C(5-Fu)and group E(P<0.05).Conclusion Silencing HIF-1αcan significantly reverse the resistance of HT-29 MCS cells to 5-Fu,VCR and ADR.