锌带基因ZNRD1在胃癌耐药细胞中的表达和功能

目的 探讨锌带基因ZNRD1在多药耐药胃癌细胞中的表达和作用。方法 应用Northern blot和半定量RT—PCR,观察锌带基因ZNRD1在胃癌细胞SGC7901及其长春新碱(VCR)诱导的耐药细胞SGC7901／VCR中的表达;将反义核酸转染SGC7901／VCR细胞,通过免疫组化检测转导细胞与非转导细胞ZNRD1蛋白的表达;以流式细胞仪检测细胞周期的变化,以MTT实验检测细胞生长曲线和对VCR、阿霉素(ADM)的药敏性。结果 胃癌耐药细胞SGC7901／VCR与非耐药细胞相比,ZNBDl基因的表达明显增高。转导株antiZNRDl—SGC7901／VCR细胞的ZNRD1蛋白水平明显低于非转导株,C1期细胞比例增加而S期比例减少,生长受到抑制,且对VCR、ADM的敏感性明显增高。结论 胃癌耐药细胞与非耐药细胞相比,ZNRDl基因处于高表达状态。反义核酸转染可有效阻断ZNRDl蛋白的表达,在一定程度上逆转人胃癌耐药细胞SGC7901／VCR的多药耐药性。

Expression and function of zinc ribbon gene ZNRD1 in drug-resistant gastric cancer cells

Objective To study the expression and function of zinc ribbon gene ZNRD1 in drug resistant cells of gastric cancer. Methods Two tumor cell lines were used in this study: gastric cancer SGC7901 and its drug resistant counterpart SGC7901/VCR stepwise selected by vincristine. The expression of ZNRD1 in SGC7901 and SGC7901/VCR was detected by northern blot and semiquantitative RT PCR. ZNRD1 antisense nucleic acid was transfected into SGC7901/VCR cells by lipofectamine. The expression of protein in SGC7901/VCR cells and the transfectants was detected by immunochemical method. Fluorescence activated cell scan (FACS) was applied to observe the cell cycle alteration. Growth curve and drug sensitization of cells for vincristine (VCR) and adriamycin (ADM) were analyzed by MTT assay. Results The expression of ZNRD1 was higher in SGC7901/VCR than in SGC7901 cells. Immunochemical results showed that the expression level of ZNRD1 protein was lower in anti ZNRD1 SGC7901/VCR cells than in non transfectants. The anti ZNRD1 SGC7901/VCR cells were gradually accumulated in G 1 phase, with a concomitant decrease of cell population in S phase. MTT assay showed that transfectant cell proliferation was lagged and more sensitive to VCR and ADM than non transfectants. Conclusion ZNRD1 gene displays high expression in VCR resistant gastric cancer cells. Expression of ZNRD1 protein is effectively blocked in anti ZNRD1 SGC7901/VCR cells by gene transfection. ZNRD1 antisense nucleic acid could reverse, to some degree, the MDR of human drug resistant gastric cancer cell SGC7901/VCR.