miR-222 can inhibit the autophagy of renal cell carcinoma cells through down-regulating the expression of DDIT4

倪晓辰; 赵志红; 马永良

doi:10.3969/j.issn.1007-3969.2015.03.001

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miR-222 can inhibit the autophagy of renal cell carcinoma cells through down-regulating the expression of DDIT4

更新时间：2025-12-31

- miR-222 can inhibit the autophagy of renal cell carcinoma cells through down-regulating the expression of DDIT4
- China Oncology Vol. 25, Issue 3, Pages: 161-166(2015)
- 作者机构：
  
  河北医科大学第四医院泌尿外科,河北,石家庄,050011
- 作者简介：
- 基金信息：
- DOI：10.3969/j.issn.1007-3969.2015.03.001
  CLC：
- Published Online：18 May 2015，
  
  Published：18 May 2015
- 稿件说明：
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倪晓辰,赵志红,马永良,等. miR-222通过下调DDIT4抑制肾癌细胞自噬[J]. 中国癌症杂志, 2015, 25(3): 161-166.

倪晓辰, 赵志红, 马永良. miR-222 can inhibit the autophagy of renal cell carcinoma cells through down-regulating the expression of DDIT4[J]. China Oncology, 2015, 25(3): 161-166.
倪晓辰,赵志红,马永良,等. miR-222通过下调DDIT4抑制肾癌细胞自噬[J]. 中国癌症杂志, 2015, 25(3): 161-166. DOI： 10.3969/j.issn.1007-3969.2015.03.001.

倪晓辰, 赵志红, 马永良. miR-222 can inhibit the autophagy of renal cell carcinoma cells through down-regulating the expression of DDIT4[J]. China Oncology, 2015, 25(3): 161-166. DOI： 10.3969/j.issn.1007-3969.2015.03.001.

摘要

背景与目的：微小RNA(microRNA，miRNA，miR)在肿瘤的发生、发展中具有重要的作用。miR-222在多种肿瘤组织中表达上调，而其在肾癌(renal cell carcinoma，RCC)中的表达及其作用机制尚不清楚。本研究拟通过检测RCC组织及相应癌旁组织中miR-222的表达情况，探讨miR-222在RCC中的作用。并在体外条件下，通过检测miR-222的下游作用靶点，探讨miR-222的抗RCC作用机制。方法：采用实时定量聚合酶链反应(quantitative real-time polymerase chain reaction，qRT-PCR)检测RCC组织和癌旁组织中miR-222的表达；采用CCK-8法检测细胞的增殖活力；采用蛋白[质]印迹法(Western blot)检测DDIT4蛋白及LC3-Ⅱ的表达；采用荧光素酶实验验证miR-222的作用靶点；转染EGFP-LC3后在激光共聚焦显微镜下观察细胞自噬状态。结果：qRTPCR检测结果显示，miR-222在RCC组织中表达明显上调。在人肾透明细胞癌786-O细胞株中敲低miR-222表达后显著抑制细胞的增殖活力，而过表达miR-222可增强786-O细胞的增殖活力(P0.01)。在786-O细胞中敲低miR-222后，其靶基因DDIT4蛋白的表达显著上调，过表达miR-222后，DDIT4表达明显下调。荧光素酶实验结果表明，DDIT4为miR-222的直接作用靶点。RCC组织的DDIT4表达下调。在786-O细胞中抑制miR-222表达后，LC3-Ⅱ的表达水平显著上调，自噬体数目显著增加。结论：miR-222在RCC中高表达，并可能通过靶向抑制DDIT4的表达参与调控RCC细胞自噬。

Abstract

Background and purpose: MicroRNA (miRNA

miR) plays an important regulatory role in cancer. miR-222 is reported to be up-regulated in various tumors

but its role in renal cell carcinoma (RCC) remains unclear. In this study

we detected the expression of miR-222 in both RCC and adjacent tissue samples. The aim of this study was to investigate the role of miR-222 in RCC. Methods: The expression levels of miR-222 in RCC tissue samples were quantified by quantitative real-time polymerase chain reaction (qRT-PCR). DDIT4 and LC3-Ⅱ protein expressions were determined by Western blot. Dual luciferase assay was performed to verify the target of miR-222. EGFP-LC3 microscopy assay was performed to assess autophagy. Results: Results from qRT-PCR showed that the expression of miR-222 was up-regulated in RCC tissues. Knockdown of miR-222 with specific antagomiR decreased the cell viability of 786-O cells

whereas overexpression of miR-222 increased the cell viability (P0.01). The levels of DDIT4 were upregulated in 786-O cells transfected with miR-222 antagomiR

whereas overexpression of miR-222 induced the downregulation of DDIT4 expression. Data from dual luciferase assay indicated that miR-222 directly targeted the expression of DDIT4. Consistently

the expression of DDIT4 in RCC tissues was down-regulated compared with adjacent tissues. Knockdown of miR-222 in 786-O cells induced a significant increase of autophagosome formation and LC3 lipidation. These results supported that miR-222 could inhibit autophagy in RCC cells

which may affect the clinical characteristcs of RCC. Conclusion: miR-222 is up-regulated in RCC and can inhibit the autophagy of RCC cells through downregulating the expression of DDIT4.

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