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首页> 外文期刊>World Journal of Gastroenterology >Effect of ZNRD1 gene antisense RNA on drug resistant gastric cancer cells
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Effect of ZNRD1 gene antisense RNA on drug resistant gastric cancer cells

机译：ZNRD1基因反义RNA对耐药胃癌细胞的影响

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AIM: To investigate the expression level of ZNRD1 gene in gastric cancer cells SGC7901 and gastric cancer MDR (multidrug resistant) cells SGC7901/VCR, and to observe the drug sensitizing and proliferation effect of ZNRD1 antisense nucleic acid transduction on SGC7901/VCR cells. METHODS: Amplification of sequences encoding ZNRD1 from SGC7901/VCR cDNA by PCR. The levels of ZNRD1 mRNA expression were demonstrated using semiquantitative reverse transcription polymerase chain reaction (RT-PCR). Eukaryotic expression vector pcDNA3.1-anti ZNRD1 was constructed and transfected into SGC7901/VCR cells by lipofectamine. Immunochemical method was used to detect the expression of protein in SGC7901/VCR cells and transfectants. The cell cycle alteration and the intracellular adriamycin (ADM) accumulation were observed by FACS. Growth curve and drug sensitization of cells for vincristine (VCR) were analyzed with MTT assay. RESULTS: We cloned the open reading frame of full-length ZNRD1. The expression of ZNRD1 showed higher in SGC7901/VCR than in SGC7901 cells. The antisense ZNRD1 drug-resistant clones were selected after gene transfection. Immunochemical results showed that the expression level of ZNRD1 protein was lower in anti ZNRD1-SGC7901/VCR cells than that in non-transfectants. Comparing to SGC7901/ VCR and pcDNA3.1-SGC7901/VCR, anti ZNRD1-SGC7901/ VCR showed gradually accumulated in G_1 phase, with a concomitant decrease of cell population in S phase. FACS also suggested intracellular ADM accumulation increased 2fold in SGC7901/VCR cells after transfected with antisense ZNRD1. MTT assay showed that transfectants cells proliferation was lagged and more sensitive to VCR than non-transfectants. CONCLUSION: ZNRD1 gene displayed highly expression in VCR resistant gastric cancer cells. Expression of ZNRD1 protein was effectively blocked in anti ZNRD1-SGC7901/VCR cells by gene transfection. ZNRD1 antisense nucleic acid transfection sensitized drug resistant gastric cancer cells to VCR, increased ADM accumulation and inhibited the cells proliferation. ZNRD1 antisense RNA transduction could reverse the MDR of human drug-resistant gastric cancer cell SGC7901/VCR to a degree.
机译：目的：研究ZNRD1基因在胃癌细胞SGC7901和胃癌MDR（多药耐药）细胞SGC7901 / VCR中的表达水平，观察ZNRD1反义核酸转导对SGC7901 / VCR细胞的敏化和增殖作用。方法：通过PCR扩增SGC7901 / VCR cDNA中编码ZNRD1的序列。使用半定量逆转录聚合酶链反应（RT-PCR）证明ZNRD1 mRNA的表达水平。构建了真核表达载体pcDNA3.1-抗ZNRD1，并通过脂质转染胺转染到SGC7901 / VCR细胞中。采用免疫化学方法检测SGC7901 / VCR细胞和转染子中蛋白质的表达。通过FACS观察细胞周期改变和细胞内阿霉素（ADM）的积累。用MTT法分析了长春新碱（VCR）的生长曲线和药物敏感性。结果：我们克隆了全长ZNRD1的开放阅读框。在SGC7901 / VCR中，ZNRD1的表达高于SGC7901细胞。基因转染后选择反义ZNRD1耐药克隆。免疫化学结果显示，ZNRD1蛋白在抗ZNRD1-SGC7901 / VCR细胞中的表达水平低于非转染株。与SGC7901 / VCR和pcDNA3.1-SGC7901 / VCR相比，抗ZNRD1-SGC7901 / VCR在G_1期逐渐积累，在S期细胞数量随之减少。 FACS还表明，用反义ZNRD1转染后，SGC7901 / VCR细胞中细胞内ADM积累增加了2倍。 MTT分析表明，转染子细胞的增殖比非转染子的细胞滞后并且对VCR更敏感。结论：ZNRD1基因在VCR耐药胃癌细胞中高表达。通过基因转染可有效阻止ZNRD1蛋白在抗ZNRD1-SGC7901 / VCR细胞中的表达。 ZNRD1反义核酸转染使耐药的胃癌细胞对VCR敏感，增加了ADM的积累并抑制了细胞的增殖。 ZNRD1反义RNA转导可在一定程度上逆转人耐药胃癌细胞SGC7901 / VCR的MDR。

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