Department of General Surgery, Caoxian People's Hospital, Heze, China.
Eur Rev Med Pharmacol Sci. 2020 Sep;24(17):8911-8917. doi: 10.26355/eurrev_202009_22832.
The purpose of this study was to detect the relative expression of long non-coding ribonucleic acid (lncRNA) in non-homologous end joining pathway 1 (LINP1) in papillary thyroid cancer (PTC) tissues and cells, and to investigate the molecular mechanisms of abnormal expression and biological function of LINP1.
The relative expression of LINP1 in PTC tissues and cells was detected via quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR), and the impact of small interfering (si)-LINP1 on the proliferative capacity of PTC cells was studied using Cell Counting Kit-8 (CCK-8) and colony formation assays. After the expression of LINP1 in PTC cells was interfered, flow cytometry was applied to determine the changes in cell cycle distribution and apoptosis rate. The transcription factors binding to the promoter region of LINP1 were predicted by bioinformatics. Next, qRT-PCR assay was adopted to measure the changes in LINP1 expression after interference in the expression of signal transducer and activator of transcription 1 (STAT1). Finally, the changes in the expressions of molecular markers of the adenosine 5'-monophosphate-activated protein kinase (AMPK) signaling pathway were examined via Western blotting assay after the expressions of STAT1 and LINP1 were interfered.
It was shown in qRT-PCR results that LINP1 expression was upregulated in 42 out of 53 cases of PTC tissues and in all PTC cells. After interference in the expression of LINP1 in PTC cells, the results of CCK-8 and colony formation assays indicated that the proliferative capacity of the cells was repressed. According to the results of flow cytometry, the cell cycle was arrested at the G1/G0 phase, and the apoptosis rate was increased. In addition, the bioinformatics predicted that STAT1 could bind to the promoter region of LINP1, and the results of qRT-PCR indicated that the expression of LINP1 declined after STAT1 expression was interfered. Moreover, it was indicated in the Western blotting assay after interference in the expressions of STAT1 and LINP1 that the expression of molecular marker (Phosphorylation AMPK, p-AMPK) of the AMPK signaling pathway was altered but the expression of total AMPK did not change.
The transcription factor STAT1 promotes the expression of LINP1 in PTC, and highly expressed LINP1 facilitates the proliferation and inhibits the apoptosis of PTC by suppressing the AMPK signaling pathway.
本研究旨在检测非同源末端连接通路 1（LINP1）在甲状腺乳头状癌（PTC）组织和细胞中的长非编码 RNA（lncRNA）的相对表达，并探讨 LINP1 异常表达的分子机制及其生物学功能。
采用实时荧光定量逆转录聚合酶链反应（qRT-PCR）检测 PTC 组织和细胞中 LINP1 的相对表达，采用细胞计数试剂盒-8（CCK-8）和集落形成实验研究小干扰（si）-LINP1 对 PTC 细胞增殖能力的影响。干扰 PTC 细胞中 LINP1 的表达后，采用流式细胞术检测细胞周期分布和凋亡率的变化。通过生物信息学预测 LINP1 启动子区域结合的转录因子。然后，采用 qRT-PCR 检测干扰信号转导和转录激活因子 1（STAT1）表达后 LINP1 表达的变化。最后，干扰 STAT1 和 LINP1 的表达后，采用 Western blot 检测腺苷酸活化蛋白激酶（AMPK）信号通路分子标记物的表达变化。
qRT-PCR 结果显示，53 例 PTC 组织中有 42 例 LINP1 表达上调，所有 PTC 细胞均表达 LINP1。干扰 PTC 细胞中 LINP1 的表达后，CCK-8 和集落形成实验结果表明，细胞增殖能力受到抑制。根据流式细胞术的结果，细胞周期被阻滞在 G1/G0 期，凋亡率增加。此外，生物信息学预测 STAT1 可与 LINP1 启动子区域结合，qRT-PCR 结果表明 STAT1 表达被干扰后 LINP1 的表达下降。此外，干扰 STAT1 和 LINP1 的表达后，Western blot 检测结果显示 AMPK 信号通路的分子标记物（磷酸化 AMPK，p-AMPK）表达发生改变，但总 AMPK 的表达没有变化。
转录因子 STAT1 促进 PTC 中 LINP1 的表达，高表达的 LINP1 通过抑制 AMPK 信号通路促进 PTC 的增殖并抑制其凋亡。
