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目的探讨Runx3基因对HepG2细胞耐药性的影响及可能机制。方法取在1.6μg/ml顺铂(CDDP)下生长良好的HepG2耐药细胞(HepG2/CDDP-1.6),建立耐药HepG2/CDDP的动态变化模型HepG2/CDDP/2.0,并予以甲基转移酶抑制剂5-氮-2’-脱氧胞苷(5-Aza-CdR)处理;将Runx3-shRNA表达载体转染HepG2/CDDP-1.6细胞,RT-PCR检测处理及转染前后细胞Runx3 mRNA的表达。MTT法检测细胞的增殖活性,流式细胞仪分析细胞周期,Hoechst 33258检测凋亡,Western blot检测P-gp表达的变化。结果在动态变化模型中随CDDP诱导时间的延长,Runx3 mRNA的表达逐渐降低。5-Aza-CdR处理后Runx3 mRNA的表达增加,细胞生长受到明显抑制,S期细胞逐渐增加,凋亡细胞明显增多,细胞内P-gp的表达减少。转染Runx3-shRNA载体后HepG2/CDDP-1.6细胞对CDDP的耐受性增强,G1期细胞增加,P-gp的表达增加。结论 Runx3基因的表达可抑制HepG2/CDDP耐药的形成,提高HepG2/CDDP细胞对化疗药物的敏感性。
Objective To investigate the effect of Runx3 gene on drug resistance of HepG2 cells and its possible mechanism. Methods HepG2 / CDDP-1.6 cells were cultured in the presence of 1.6μg / ml cisplatin (CDDP). The HepG2 / CDDP / 2.0 cell line was established and treated with methyltransferase The expression of Runx3 mRNA in HepG2 / CDDP-1.6 cells transfected with Runx3-shRNA expression vector was detected by RT-PCR and the expression of Runx3-shRNA was detected by RT-PCR and 5-Aza-CdR treatment. . Cell proliferation was detected by MTT assay, cell cycle was analyzed by flow cytometry, apoptosis was detected by Hoechst 33258, and P-gp expression was detected by Western blot. Results The expression of Runx3 mRNA gradually decreased with the increase of CDDP induction time in the dynamic model. After 5-Aza-CdR treatment, the expression of Runx3 mRNA increased, the cell growth was significantly inhibited, the S phase cells gradually increased, the apoptotic cells increased significantly, and the expression of P-gp decreased. After transfected with Runx3-shRNA vector, the resistance of HepG2 / CDDP-1.6 cells to CDDP was enhanced, and the cells in G1 phase increased and the expression of P-gp increased. Conclusion The expression of Runx3 gene can inhibit the formation of HepG2 / CDDP and increase the sensitivity of HepG2 / CDDP cells to chemotherapeutic drugs.