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Hsa_circ_0035483 通过自噬调控海绵吸附 hsa-miR-335 促进人肾癌细胞对吉西他滨的耐药性。

Hsa_circ_0035483 sponges hsa-miR-335 to promote the gemcitabine-resistance of human renal cancer cells by autophagy regulation.

机构信息

Department of Nephrology, Zhengzhou University People's Hospital, Henan Provincial People's Hospital, China.

School of Physical Education, Wuhan Business University, China.

出版信息

Biochem Biophys Res Commun. 2019 Oct 29;519(1):172-178. doi: 10.1016/j.bbrc.2019.08.093. Epub 2019 Sep 3.

DOI:10.1016/j.bbrc.2019.08.093

PMID:31492499

Abstract

Renal clear cell carcinoma (RCC) is the most common pathological type of renal carcinoma and drug resistance often occurs. We studied the effect of hsa_circ_0035483 on gemcitabine sensitivity in RCC, and explored its regulatory effect on downstream hsa-miR-335 and Cyclin B1 (CCNB1). High-throughput sequencing was used to analyze the differentially expressed circRNA in RCC. The expressions of hsa_circ_0035483, hsa-miR-335, CCNB1, and autophagy-related proteins were detected by RT-PCR or Western blot. The target relationships were revealed by RNA pulldown assay and dual luciferase report assay. Autophagy marker LC3 was detected by immunofluorescence. Cell viability was detected by MTT assay. Hsa_circ_0035483 can facilitate gemcitabine-induced autophagy, and enhance the resistance of RCC to gemcitabine. Hsa-miR-335 is the target regulatory point of hsa_circ_0035483. In addition, hsa_circ_0035483 promotes autophagy and tumor growth and enhances gemcitabine resistance in RCC by regulating hsa-miR-335/CCNB1, and silenced hsa_circ_0035483 can enhance gemcitabine sensitivity in vivo. Hsa_circ_0035483 may be the target of gemcitabine resistance in the treatment of RCC.

摘要

肾透明细胞癌（RCC）是最常见的肾细胞癌病理类型，常发生耐药。我们研究了 hsa_circ_0035483 对 RCC 吉西他滨敏感性的影响，并探讨了其对下游 hsa-miR-335 和细胞周期蛋白 B1（CCNB1）的调节作用。利用高通量测序分析 RCC 中差异表达的 circRNA。通过 RT-PCR 或 Western blot 检测 hsa_circ_0035483、hsa-miR-335、CCNB1 和自噬相关蛋白的表达。通过 RNA 下拉实验和双荧光素酶报告实验揭示靶标关系。通过免疫荧光检测自噬标志物 LC3。通过 MTT 法检测细胞活力。hsa_circ_0035483 可促进吉西他滨诱导的自噬，增强 RCC 对吉西他滨的耐药性。hsa-miR-335 是 hsa_circ_0035483 的靶标调控点。此外，hsa_circ_0035483 通过调节 hsa-miR-335/CCNB1 促进自噬和肿瘤生长，并增强 RCC 对吉西他滨的耐药性，沉默 hsa_circ_0035483 可增强体内吉西他滨的敏感性。hsa_circ_0035483 可能是治疗 RCC 中吉西他滨耐药的靶点。