Silencing AREG Enhances Sensitivity to Irradiation by Suppressing the PI3K/AKT Signaling Pathway in Colorectal Cancer Cells.

IF 5.3 Q1 MEDICINE, RESEARCH & EXPERIMENTAL Biologics : Targets & Therapy Pub Date : 2024-09-28 eCollection Date: 2024-01-01 DOI:10.2147/BTT.S480361
Wenbing Zhang, Wenjuan Zhang, Chenling Tang, Yan Hu, Ke Yi, Xiaohui Xu, Zhihua Chen
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Abstract

Background: It has been established that Spalt-Like Transcription Factor 4 (SALL4) promotes Colorectal Cancer (CRC) cell proliferation. Furthermore, Amphiregulin (AREG) is crucially involved in cancer cell proliferation and therapeutic resistance regulation. In this regard, this study aimed to establish whether SALL4 affects the radiosensitization of CRC cells via AREG expression regulation.

Methods: Transcriptome sequencing and the Human Transcription Factor Database (HumanTFDB) were used to identify the potential SALL4 targets. The dual-luciferase reporter analysis was used to confirm the SALL4-induced AREG activation. Western Blot (WB) and Reverse Transcription quantitative Polymerase Chain Reaction (RT-qPCR) assays were used to examine the effect of X-ray irradiation on SALL4 and AREG expression. The AREG-KD (Knockdown) stable cell lines were created through lentiviral infection. Cell proliferation was tracked using Cell Counting Kit 8 (CCK-8) and 5-Ethynyl-2'-deoxyuridine (EdU)-incorporation assays. Cell cycle and apoptosis were examined through flow cytometry. The cells were exposed to a controlled X-ray radiation dose (6 Gy) for imaging purposes.

Results: SALL4 could bound to the AREG promoter, enhancing AREG expression. Furthermore, irradiation upregulated SALL4 and AREG in CRC cells. Additionally, AREG knockdown in CRC cells led to reduced DNA replication efficiency, suppressed cell proliferation, increased DNA damage, and enhanced G1 phase arrest and apoptosis following irradiation. On the other hand, AREG overexpression reversed the inhibitory effects of SALL4 downregulation on AREG expression.

Conclusion: In CRC cells, SALL4 downregulation suppressed AREG expression, regulating CRC cell radiosensitivity via the PI3K-AKT pathway, thus presenting a potential therapeutic pathway for CRC treatment using Radiotherapy (RT).

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抑制 AREG 可抑制结直肠癌细胞的 PI3K/AKT 信号通路,从而提高其对辐照的敏感性
背景:Spalt-Like 转录因子 4(SALL4)可促进结直肠癌(CRC)细胞增殖。此外,安非他酮(Amphiregulin,AREG)在癌细胞增殖和抗药性调控中起着至关重要的作用。因此,本研究旨在确定SALL4是否通过AREG表达调控影响CRC细胞的放射增敏:方法:利用转录组测序和人类转录因子数据库(HumanTFDB)确定SALL4的潜在靶点。双荧光素酶报告分析用于证实 SALL4 诱导的 AREG 激活。Western Blot(WB)和反转录定量聚合酶链反应(RT-qPCR)检测了X射线照射对SALL4和AREG表达的影响。通过慢病毒感染建立了 AREG-KD (敲除)稳定细胞系。使用细胞计数试剂盒8(CCK-8)和5-乙炔基-2'-脱氧尿苷(EdU)-incorporation检测法跟踪细胞增殖。通过流式细胞仪检测细胞周期和细胞凋亡。为成像目的,细胞暴露于可控的 X 射线辐射剂量(6 Gy):结果:SALL4能与AREG启动子结合,从而增强AREG的表达。此外,辐照可上调 SALL4 和 AREG 在 CRC 细胞中的表达。此外,在 CRC 细胞中敲除 AREG 会导致 DNA 复制效率降低、细胞增殖受抑制、DNA 损伤增加以及照射后 G1 期停滞和细胞凋亡增强。另一方面,AREG的过表达逆转了SALL4下调对AREG表达的抑制作用:结论:在CRC细胞中,SALL4下调抑制了AREG的表达,通过PI3K-AKT途径调节CRC细胞的放射敏感性,从而为利用放射治疗(RT)治疗CRC提供了一条潜在的治疗途径。
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来源期刊
Biologics : Targets & Therapy
Biologics : Targets & Therapy MEDICINE, RESEARCH & EXPERIMENTAL-
CiteScore
8.30
自引率
0.00%
发文量
22
审稿时长
16 weeks
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