PAX5 激活的长链非编码 RNA UASR1 通过调节 mTOR 通路促进结直肠癌的生长。
Promoting effect of PAX5-activated lncRNA UASR1 on growth of colorectal cancer by regulating the mTOR pathway.
机构信息
Department of Colorectal Surgery, Changhai Hospital, Shanghai, China.
出版信息
Eur Rev Med Pharmacol Sci. 2020 Mar;24(6):2986-2993. doi: 10.26355/eurrev_202003_20663.
OBJECTIVE
To study the expression of long non-coding ribonucleic acid (lncRNA) UNC5B antisense RNA 1 (UASR1) in colorectal cancer (CRC) and its biological functions, and to discuss the regulatory effect of the transcription factor on lncRNA UASR1.
PATIENTS AND METHODS
The expressions of lncRNA UASR1 in the CRC tissues and cells were detected via quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) assay. After the expression of lncRNA UASR1 was interfered, the change in the CRC cell proliferation ability was investigated through cell counting kit-8 (CCK-8) assay and colony formation assay, respectively. Changes in cell cycle distribution and apoptosis rate in CRC cells after transfection of small-interfering UASR1 (si-UASR1) were detected using flow cytometry. Potential transcription factors binding UASR1 promoter region were analyzed through bioinformatics. The change in the UASR1 expression was measured through the qRT-PCR assay after the paired box 5 (PAX5) expression was interfered. Following the interference in the expressions of PAX5 and UASR1, expression changes in the molecular markers of the mammalian target of rapamycin (mTOR) signaling pathway were detected via Western blotting assay.
RESULTS
The qRT-PCR results indicated that the UASR1 expression was up-regulated in 39/45 CRC tissues, and it identically presented the up-regulated expression level in the CRC cells. After the UASR1 expression was interfered, the CRC cell proliferation ability was degraded according to the CCK-8 assay and colony formation assay. Based on the flow cytometry results, compared with the small-interfering-negative control (si-NC) group, the cell cycle was arrested in the G1/G0 phase in the si-UASR1 group, and apoptosis rate increased. Bioinformatics and qRT-PCR results showed that the transcription factor PAX5 regulated the UASR1 expression. The Western blotting assay indicated that the expressions of the molecular markers of the mTOR signaling pathway were changed after the expressions of PAX5 and UASR1 were interfered.
CONCLUSIONS
The transcription factor PAX5 promotes the expression of lncRNA UASR1 in CRC. The highly expressed UASR1 facilitates the malignant proliferation of CRC via the mTOR signaling pathway.
目的
研究长链非编码核糖核酸（lncRNA）UNC5B 反义 RNA 1（UASR1）在结直肠癌（CRC）中的表达及其生物学功能，并探讨转录因子对 lncRNA UASR1 的调控作用。
患者和方法
采用实时定量逆转录聚合酶链反应（qRT-PCR）检测 CRC 组织和细胞中 lncRNA UASR1 的表达。干扰 lncRNA UASR1 的表达后，分别通过细胞计数试剂盒-8（CCK-8）检测和集落形成实验检测 CRC 细胞增殖能力的变化。用流式细胞术检测转染小干扰 UASR1（si-UASR1）后 CRC 细胞周期分布和凋亡率的变化。通过生物信息学分析预测 UASR1 启动子区域的潜在转录因子。干扰配对盒 5（PAX5）的表达后，通过 qRT-PCR 检测 UASR1 的表达变化。干扰 PAX5 和 UASR1 的表达后，通过 Western blot 检测哺乳动物雷帕霉素靶蛋白（mTOR）信号通路的分子标志物的表达变化。
结果
qRT-PCR 结果表明，39/45 例 CRC 组织中 UASR1 表达上调，CRC 细胞中同样呈现上调表达水平。干扰 UASR1 表达后，CCK-8 检测和集落形成实验均显示 CRC 细胞增殖能力下降。根据流式细胞术结果，与小干扰阴性对照（si-NC）组相比，si-UASR1 组细胞周期停滞在 G1/G0 期，凋亡率增加。生物信息学和 qRT-PCR 结果表明，转录因子 PAX5 调控 UASR1 的表达。Western blot 检测结果表明，干扰 PAX5 和 UASR1 的表达后，mTOR 信号通路的分子标志物表达发生变化。
结论
转录因子 PAX5 促进 CRC 中 lncRNA UASR1 的表达。高表达的 UASR1 通过 mTOR 信号通路促进 CRC 的恶性增殖。
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