The Inhibition of Venin Arginine Esterase Agkihpin in the Expression of MRP1 in the SMMC-7721 Cell Strain
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摘要:
目的 探讨蛇毒精氨酸酯酶Agkihpin对人肝癌细胞株SMMC-7721中多药耐药相关蛋白1(multidrug resistance associated protein 1, MRP1)表达的影响, 并阐明Agkihpin抑制人肝癌SMMC-7721细胞活力的机制。 方法 目的: 方法: 采用不同浓度的Agkihpin处理SMMC-7721细胞72 h后, 应用免疫细胞化学、Western blot和RT-PCR等方法检测MRP1在SMMC-7721细胞中的转录和表达。 结果 不同浓度Agkihpin作用SMMC-7721细胞72 h后MRP1表达均降低, 显示Agkihpin可显著下调SMMC-7721细胞中MRP1转录和表达(P < 0.05), 并呈现出一定的浓度依赖效应。 结论 Agkibpin能抑制人低分化肝癌细胞株SMMC-7721中MRP1的表达, 并呈一定程度的浓度依赖效应, 提示Agkihpin在一定程度上可用于提高肝癌细胞对化疗药物的敏感性。 Abstract:Objective To explore the effects of Agkihpin on the expression of multidrug resistance-associated protein 1(MRP1) in the SMMC-7721 cell line and to describe the mechanism of SMMC-7721 inhibition by Agkihpin. Methods The cultured cells were treated with different concentrations of Agkihpin for 72 h.The transcription and expression levels of MRP 1 in SMMC-7721 were assayed using immunocytochemistry, Western blot, and reverse transcription-polymerase chain reaction. Results Compared with the control group, both the transcription and the expression levels of MRP1 in SMMC-7721 were significantly reduced after treatment with different concentrations of Agkihpin(P < 0.05) in a concentration-dependent manner. Conclusion Agkihpin can inhibit the transcription and expression of MRP1 in SMMC-7721 cells, which is likely responsible for the inhibition of cellular vitality and, to some extent, the enhancement of the sensitivity of the hepatocellular carcinoma cells to chemotherapeutics. -
图 1 免疫细胞化学检测不同处理组SMMC-7721细胞MRP1蛋白表达(×400)
A：为阴性对照，其中PBS代替一抗；B~D：Agkihpin终浓度分别为0、1.033及4.13 μg/mL，棕黄色阳性颗粒染色主要位于胞膜和胞质
Figure 1. Immunocytochemistry shows the expression of the MRP1 protein in SMMC-7721 cells treated with different concentrations of Agkihpin for 72 h(×400)
图 2 不同处理组SMMC-7721细胞MRP1的转录和表达
A：RT-PCR检测；B：Western blot检测
1~8中Agkihpin终浓度分别为0、0.207、0.259、0.413、0.516、1.033、
2.065及4.13 μg/mL；M为DNA分子标记Figure 2. Transcription and expression of MRP1 in SMMC-7721 cells treated with different concentrations of Agkihpin
图 3 不同浓度Agkinpin处理组SMMC-7721细胞MRP1的转录和表达下调率的比较
Figure 3. Comparison of down-regulated rates of transcription and expression of MRP1 among groups of SMMC-7721 cells treated with different concentrations of Agkihpin
表 1 不同浓度Agkihpin作用后SMMC-7721细胞MRP1转录和表达强度比较 （x±s）
Table 1. Effects of Agkihpin on the transcription and expression intensities of MRP1 in SMMC-7721 cells (x±s)
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