沉默ABCC4基因逆转人胃癌多药耐药的研究

Silencing ABCC4 Gene Reverses Multi-drug Resistance of Human Gastric Cancer Cells

目的探讨胃癌耐药复发相关基因ABCC4在胃癌耐药复发中的分子机制。方法采用免疫组织化学、Western blot、RT-PCR、流式细胞检测胃癌组织及胃癌耐药细胞中ABCC4的表达情况,利用RNA干扰技术下调ABCC4基因在胃癌耐药细胞中的表达水平。结果 ABCC4基因在多种胃癌细胞中高表达,尤其是在胃癌耐药细胞中表达增加更为显著,但在正常胃黏膜细胞中极低表达或表达缺失。RNA干扰下调ABCC4基因可导致胃癌耐药细胞发生凋亡,并且细胞周期被阻滞在G1期以内。5-Fu可以显著抑制ABCC4沉默后肿瘤细胞增殖,同时,也能明显抑制ABCC4沉默后裸鼠体内肿瘤生长。结论 ABCC4基因过度表达于胃癌耐药细胞中,下调ABCC4基因表达可抑制胃癌多药耐药细胞增殖,并且可增强其对化疗药物的敏感度。

Objective To investigate the molecular mechanism of ABCC4 gene in the recurrence of drug resistance in gastric cancer.Methods ABCC4 expression in gastric cancer tissue and multi-drug resistant gastric cancer cells were detected by immunohistochemistry,Western blot,RT-PCR and flow cytometry.The expression level of ABCC4 was down-regulated by RNA interference in drug-resistant gastric cancer cells.Results ABCC4 was highly expressed in variety of cancer cells,especially in drug-resistant gastric cancer cells,however,it was lowly or deficiently expressed in normal gastric mucosa cells.Down-regulating ABCC4 by RNA interference could lead to the apoptosis of drug-resistant gastric cancer cells and the block of cell cycle at G1 phase.5-Fu could significantly inhibit the proliferation of tumor cells after ABCC4 were silenced,at the same time,it could also suppress the growth of tumor in nude mice.Conclusion ABCC4 was overexpressed in drug-resistant gastric cancer cells.Down-regulating ABCC4 expression by RNA interference could inhibit the proliferation of multi-drug resistant gastric cancer cells,and strengthen their sensitivity to chemotherapy.