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Circ-NT5C2 stimulates FZD4 expression to promote the malignant progression of osteosarcoma by targeting miR-488-3p
Applied Biological Chemistry ( IF 3.2 ) Pub Date : 2022-10-31 , DOI: 10.1186/s13765-022-00735-5
Yang, Xiaoqi, Wang, Shuhua, Zhang, Xianjun, Gao, Xiangbin, Xu, Pengfei

Circ-NT5C2 has been confirmed to be highly expressed and associated to the progression of osteosarcoma (OS). However, the behind mechanism of circ-NT5C2 involvement in OS remains unclear. The expression of circ-NT5C2, miR-488-3p and FZD4 was measured by quantitative real-time PCR, and the protein expression of E-cadherin, N-cadherin and FZD4 was detected by western blot. Cell counting kit 8 assay, colony formation assay and 5-ethynyl-2-deoxyuridine assay were performed to assess the cell proliferation. The cell apoptosis was measured by flow cytometry and Caspase3/Caspase9 Activity Assay Kits. Cell migration and invasion were detected by transwell assay. Dual-luciferase reporter assay and RIP assay were carried out to determine the binding relation among circ-NT5C2, miR-488-3p and FZD4. Animal experiment and immunohistochemistry analysis were conducted to explore the role of circ-NT5C2 in tumor growth in vivo. Comparing with controls, the expression of circ-NT5C2 and FZD4 was upregulated and miR-488-3p expression was downregulated in OS tumor tissues and cells. Circ-NT5C2 overexpression facilitated the cell proliferation and motility and induced cell apoptosis of OS cells, whereas circ-NT5C2 knockdown had the opposite effect. Besides, we also found and confirmed that circ-NT5C2 regulated cell malignant behaviors via modulating miR-488-3p/FZD4 axis in OS. Moreover, circ-NT5C2 silencing repressed the growth of xenografts in vivo. Circ-NT5C2 upregulated FZD4 expression via sponging miR-488-3p, thus facilitating cell malignant behaviors in OS.

中文翻译:

Circ-NT5C2通过靶向miR-488-3p刺激FZD4表达促进骨肉瘤恶性进展

Circ-NT5C2 已被证实高度表达并与骨肉瘤 (OS) 的进展相关。然而,circ-NT5C2 参与 OS 的背后机制仍不清楚。实时定量PCR检测circ-NT5C2、miR-488-3p和FZD4的表达,western blot检测E-cadherin、N-cadherin和FZD4的蛋白表达。进行细胞计数试剂盒8测定、集落形成测定和5-乙炔基-2-脱氧尿苷测定以评估细胞增殖。通过流式细胞术和 Caspase3/Caspase9 活性检测试剂盒测量细胞凋亡。通过transwell法检测细胞迁移和侵袭。进行双荧光素酶报告基因测定和RIP测定以确定circ-NT5C2、miR-488-3p和FZD4之间的结合关系。进行动物实验和免疫组化分析,探讨circ-NT5C2在体内肿瘤生长中的作用。与对照组相比,在 OS 肿瘤组织和细胞中 circ-NT5C2 和 FZD4 的表达上调,miR-488-3p 的表达下调。Circ-NT5C2 过表达促进了 OS 细胞的细胞增殖和运动并诱导细胞凋亡,而 circ-NT5C2 敲低则具有相反的效果。此外,我们还发现并证实了 circ-NT5C2 通过调节 OS 中的 miR-488-3p/FZD4 轴来调节细胞恶性行为。此外,circ-NT5C2 沉默抑制了体内异种移植物的生长。Circ-NT5C2 通过海绵化 miR-488-3p 上调 FZD4 表达,从而促进 OS 中的细胞恶性行为。OS肿瘤组织和细胞中circ-NT5C2和FZD4表达上调,miR-488-3p表达下调。Circ-NT5C2 过表达促进了 OS 细胞的细胞增殖和运动并诱导细胞凋亡,而 circ-NT5C2 敲低则具有相反的效果。此外,我们还发现并证实了 circ-NT5C2 通过调节 OS 中的 miR-488-3p/FZD4 轴来调节细胞恶性行为。此外,circ-NT5C2 沉默抑制了体内异种移植物的生长。Circ-NT5C2 通过海绵化 miR-488-3p 上调 FZD4 表达,从而促进 OS 中的细胞恶性行为。OS肿瘤组织和细胞中circ-NT5C2和FZD4表达上调,miR-488-3p表达下调。Circ-NT5C2 过表达促进了 OS 细胞的细胞增殖和运动并诱导细胞凋亡,而 circ-NT5C2 敲低则具有相反的效果。此外,我们还发现并证实了 circ-NT5C2 通过调节 OS 中的 miR-488-3p/FZD4 轴来调节细胞恶性行为。此外,circ-NT5C2 沉默抑制了体内异种移植物的生长。Circ-NT5C2 通过海绵化 miR-488-3p 上调 FZD4 表达,从而促进 OS 中的细胞恶性行为。而 circ-NT5C2 敲低具有相反的效果。此外,我们还发现并证实了 circ-NT5C2 通过调节 OS 中的 miR-488-3p/FZD4 轴来调节细胞恶性行为。此外,circ-NT5C2 沉默抑制了体内异种移植物的生长。Circ-NT5C2 通过海绵化 miR-488-3p 上调 FZD4 表达,从而促进 OS 中的细胞恶性行为。而 circ-NT5C2 敲低具有相反的效果。此外,我们还发现并证实了 circ-NT5C2 通过调节 OS 中的 miR-488-3p/FZD4 轴来调节细胞恶性行为。此外,circ-NT5C2 沉默抑制了体内异种移植物的生长。Circ-NT5C2 通过海绵化 miR-488-3p 上调 FZD4 表达,从而促进 OS 中的细胞恶性行为。
更新日期:2022-10-31
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