| 谢 波,叶文鑫,舒 彦.GCLC对肾细胞癌的生长促进作用及其上游调控机制[J].肿瘤学杂志,2019,25(8):717-722. |
| GCLC对肾细胞癌的生长促进作用及其上游调控机制 |
| Effect of Glutamate-cystein Ligase Catalytic Subunit(GCLC) on Growth of Renal Cell Carcinoma and Its Mechanism |
| 投稿时间：2018-01-29 |
| DOI：10.11735/j.issn.1671-170X.2019.08.B008 |
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| 中文关键词: GCLC 肾细胞癌 microRNA GSH |
| 英文关键词:GCLC renal cell carcinoma microRNA GSH |
| 基金项目:浙江医药卫生科技计划项目（2018KY032） |
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| 中文摘要: |
| 摘 要：［目的］ 探讨GCLC基因在肾细胞癌（RCC）中的表达量、生物学作用，以及microRNA（miRNA）对于GCLC的转录后调控。［方法］ 定量RT-PCR量化GCLC在RCC临床标本中的表达量，CCK-8和平板克隆形成实验评估GCLC对于细胞增殖的影响，过氧化物酶和DAB法用于免疫组织化学分析。荧光素酶双报告系统、定量RT-PCR和Western印迹确定miR-362-3p的直接作用靶点。［结果］ 相比癌旁组织，GCLC在RCC中的表达量显著上调（25例患者中有22例呈表达上调模式）。通过小干扰RNA（siRNA）敲低GCLC的表达量可以显著降低2GSH/GSSG的比例（P<0.05）。降低GCLC的表达量可以抑制RCC细胞的增殖和集落形成（P均<0.05）。GCLC基因在RCC细胞中是miR-362-3p的一个直接调控靶点。［结论］ GCLC基因是RCC的一个潜在致癌基因并直接受到miR-362-3p的调控。因此，敲低GCLC的表达有望成为一种新型的RCC治疗手段。 |
| 英文摘要: |
| Abstract：［Objective］ To investigate the expression and biological role of glutamate-cystein ligase catalytic subunit(GCLC) in renal cell carcinoma(RCC) and its mechanism. ［Methods］ The expression of GCLC mRNA in 25 specimens of RCC tissues and corresponding normal renal tissues was detected by quantitative RT-PCR(qRT-PCR). The RCC 786-O and ACHN cells were transfected with GCLC siRNA. Cell proliferation was examined by CCK-8 assay and colony formation assay. The expression of GCLC was detected by immunohistochemistry peroxidase and DAB methods. The miR-362-3p target was determined by luciferase reporter assays，qRT-PCR and Western blotting. ［Results］ The expression of GCLC was up-regulated in RCC tissue compared with adjacent non-tumor tissues. Knockdown of GCLC expression by siRNA in RCC cells significantly decreased the 2GSH/GSSG ratio(P<0.05). Silencing GCLC expression also inhibited proliferation and colony formation of RCC cells(all P<0.05). GCLC was identified as a direct target of miR-362-3p in RCC cells. ［Conclusion］ GCLC is a potential tumor promoter in RCC and is directly regulated by miR-362-3p. The knockdown of GCLC might be a novel therapeutic approach for RCC treatment. |
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