长链非编码 RNA Linc00210 通过海绵吸附 miR-16-5p/PTK2 轴促进非小细胞肺癌进展。
Long noncoding RNA Linc00210 promotes non-small cell lung cancer progression via sponging miR-16-5p/PTK2 axis.
机构信息
Department of Respiration, Shangluo Central Hospital, Shangluo, Shanxi, China.
出版信息
Eur Rev Med Pharmacol Sci. 2020 Sep;24(18):9438-9452. doi: 10.26355/eurrev_202009_23029.
OBJECTIVE
Long noncoding RNAs (lncRNAs) are important regulators involved in a variety of cancer development. However, the role of Linc00210 in non-small cell lung cancer (NSCLC) remains unknown. This study aims to investigate the clinical value of Linc00210 in NSCLC patients and the biological functions of Linc00210 in NSCLC.
PATIENTS AND METHODS
Gene expression in NSCLC tissues and cell lines was detected using qRT-PCR or Western blot. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and colony formation assays were conducted to evaluate the effect of Linc00210 on NSCLC cell proliferation. Transwell assay and annexin V-Fluorescein 5-isothiocyanate (FITC)/Propidium Iodide (PI) were done to analyze the effect of Linc00210 on cancer cell invasion and apoptosis, respectively. Luciferase reporter assay and RIP assay were performed to determine the target of Linc00210 and miR-16-5p. Besides, these assays were used to determine reciprocally inhibition of each other-controlled NSCLC cell behaviors. In vivo tumorigenesis experiments were applied to exhibit subcutaneous tumor growth.
RESULTS
Linc0021 was highly expressed in NSCLC tissues and cell lines. Knockdown of Linc00210 inhibited cancer cell proliferation and invasion, and increased cell apoptosis, and regulated the expression of Cyclin A1, proliferating cell nuclear antigen (PCNA), E-cadherin, N-cadherin, Bax, and Bcl-2 in NSCLC cells. Further data showed Linc00210 bound to and directly modulated the miR-16-5p levels. Impressively, overexpression of miR-16-5p suppressed NSCLC cell proliferation and invasion, but increased cell apoptosis, and these behaviors could be overturned by overexpression of Linc00210 in vitro and in vivo. Finally, Linc00210 and miR-16-5p cooperatively controlled expression of protein tyrosine kinase 2 (PTK2), a miR-16-5p target.
CONCLUSIONS
Linc00210/miR-16-5p/PTK2 signaling suggests a promising novel strategy for anti-NSCLC therapy.
目的
长链非编码 RNA（lncRNAs）是参与多种癌症发展的重要调控因子。然而，Linc00210 在非小细胞肺癌（NSCLC）中的作用尚不清楚。本研究旨在探讨 Linc00210 在 NSCLC 患者中的临床价值以及 Linc00210 在 NSCLC 中的生物学功能。
方法
采用 qRT-PCR 或 Western blot 检测 NSCLC 组织和细胞系中的基因表达。通过 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四唑溴盐（MTT）和集落形成实验评估 Linc00210 对 NSCLC 细胞增殖的影响。Transwell 实验和 Annexin V-Fluorescein 5-异硫氰酸酯（FITC）/碘化丙啶（PI）分别用于分析 Linc00210 对癌细胞侵袭和凋亡的影响。荧光素酶报告基因实验和 RIP 实验用于确定 Linc00210 和 miR-16-5p 的靶标。此外，还进行了这些实验来确定彼此相互抑制的 NSCLC 细胞行为。体内肿瘤发生实验用于显示皮下肿瘤生长。
结果
Linc00210 在 NSCLC 组织和细胞系中高表达。敲低 Linc00210 抑制了癌症细胞的增殖和侵袭，增加了细胞凋亡，并调节了 NSCLC 细胞中细胞周期蛋白 A1、增殖细胞核抗原（PCNA）、E-钙黏蛋白、N-钙黏蛋白、Bax 和 Bcl-2 的表达。进一步的数据表明，Linc00210 与 miR-16-5p 结合并直接调节其水平。令人印象深刻的是，miR-16-5p 的过表达抑制了 NSCLC 细胞的增殖和侵袭，但增加了细胞凋亡，并且这些行为可以在体外和体内通过过表达 Linc00210 来逆转。最后，Linc00210 和 miR-16-5p 协同控制蛋白酪氨酸激酶 2（PTK2）的表达，PTK2 是 miR-16-5p 的靶标。
结论
Linc00210/miR-16-5p/PTK2 信号提示了一种有前途的 NSCLC 治疗新策略。
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