环状 RNA-RNF111 通过 miR-140-5p 升高 E2F3 表达促进乳腺癌对紫杉醇耐药。
Circ-RNF111 contributes to paclitaxel resistance in breast cancer by elevating E2F3 expression via miR-140-5p.
机构信息
Department of General Surgery, China-Japan Union Hospital of Jilin University, Changchun, China.
出版信息
Thorac Cancer. 2020 Jul;11(7):1891-1903. doi: 10.1111/1759-7714.13475. Epub 2020 May 23.
BACKGROUND
Circular RNAs (circRNAs) have been demonstrated to act as key regulators in the chemoresistance of human cancers, including breast cancer (BC). Here, we aimed to explore the role of circ-RNF111 in paclitaxel (PTX) resistance of BC.
METHODS
Quantitative real-time polymerase chain reaction (qRT-PCR) was employed to determine the expression of circ-RNF111, microRNA-140-5p (miR-140-5p) and E2F transcription factor 3 (E2F3) mRNA. The half maximal inhibitory concentration (IC ) of PTX, cell viability, colony formation and cell invasion were assessed by cell counting kit-8 (CCK-8) assay, colony formation assay and transwell assay, respectively. Glucose consumption and lactate production were determined by specific kits. A murine xenograft model was established to investigate the role of circ-RNF111 in PTX resistance of BC in vivo. Dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were performed to verify the relationship between miR-140-5p and circ-RNF111 or E2F3. Western blot assay was conducted to examine the protein level of E2F3.
RESULTS
Circ-RNF111 was upregulated in PTX-resistant BC tissues and cells. Circ-RNF111 knockdown restrained IC of PTX, cell viability, colony numbers, cell invasion and glycolysis in PTX-resistant BC cells in vitro and enhanced PTX sensitivity in vivo. MiR-140-5p was a target of circ-RNF111 and miR-140-5p expression was negatively correlated with circ-RNF111 expression in BC tissues. The effect of circ-RNF111 knockdown on PTX resistance was rescued by miR-140-5p deletion. Additionally, miR-140-5p could interact with E2F3 and negatively regulate E2F3 expression. Moreover, miR-140-5p suppressed IC of PTX, cell viability, colony numbers, cell invasion and glycolysis by targeting E2F3.
CONCLUSIONS
Circ-RNF111 improved PTX resistance of BC by upregulating E2F3 via sponging miR-140-5p.
背景
环状 RNA（circRNAs）已被证明在人类癌症（包括乳腺癌）的化疗耐药中起关键调控作用。在这里，我们旨在探讨环状 RNA 结合蛋白 111（circ-RNF111）在紫杉醇（PTX）耐药乳腺癌中的作用。
方法
采用实时定量聚合酶链反应（qRT-PCR）检测 circ-RNF111、微小 RNA-140-5p（miR-140-5p）和 E2F 转录因子 3（E2F3）mRNA 的表达。通过细胞计数试剂盒-8（CCK-8）检测紫杉醇的半抑制浓度（IC ）、细胞活力、集落形成和细胞侵袭，分别通过集落形成试验和 Transwell 试验进行检测。通过特定试剂盒测定葡萄糖消耗和乳酸生成。建立了小鼠异种移植模型，以研究 circ-RNF111 在体内对 BC 紫杉醇耐药的作用。通过双荧光素酶报告基因检测和 RNA 免疫沉淀（RIP）检测验证 miR-140-5p 与 circ-RNF111 或 E2F3 的关系。通过 Western blot 检测 E2F3 蛋白水平。
结果
PTX 耐药的 BC 组织和细胞中 circ-RNF111 上调。circ-RNF111 敲低抑制了 PTX 耐药的 BC 细胞中 PTX 的 IC 、细胞活力、集落数量、细胞侵袭和糖酵解作用，并增强了体内的 PTX 敏感性。miR-140-5p 是 circ-RNF111 的靶基因，BC 组织中 circ-RNF111 的表达与 miR-140-5p 的表达呈负相关。circ-RNF111 敲低对 PTX 耐药的影响可以通过 miR-140-5p 的缺失来挽救。此外，miR-140-5p 可以与 E2F3 相互作用，并负调控 E2F3 的表达。此外，miR-140-5p 通过靶向 E2F3 抑制 PTX 的 IC 、细胞活力、集落数量、细胞侵袭和糖酵解作用。
结论
circ-RNF111 通过海绵吸附 miR-140-5p 上调 E2F3 来提高 BC 的 PTX 耐药性。
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