Role of telomeric RAP1 in radiation sensitivity modulation and its interaction with CSC marker KLF4 in colorectal cancer
Role of telomeric RAP1 in radiation sensitivity modulation and its interaction with CSC marker KLF4 in colorectal cancer
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DOI：
10.1080/09553002.2020.1721609
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发表时间：
2020-02-11
影响因子：
2.6
通讯作者：
Banerjee, Birendranath
中科院分区：
文献类型：
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作者：
Anuja, Kumari;Kar, Madhabananda;Banerjee, Birendranath
目的：放射治疗是治疗结直肠癌局部肿瘤的主要手段之一。肿瘤干细胞（cancer stem cells，CSCs）的放射耐受性是肿瘤治疗的主要障碍。了解CSC的抗辐射特性可能有助于实现靶向放射治疗和提高无病生存率。端粒RAP 1参与调节多种转录因子，导致异常细胞增殖和肿瘤细胞迁移。因此，我们研究了RAP 1在维持辐射抗性细胞的抗性表型和获得性干细胞中的作用。主要方法：通过细胞存活率和DNA损伤谱分析对HCT 116来源的放射抗性细胞（HCT 116 RR）进行表征。照射后通过Western印迹和Real-time PCR研究RAP 1沉默细胞的DNA损伤和CSC标志物的表达。采用分子对接和免疫共沉淀技术研究RAP 1与KLF 4的相互作用，并分析RAP 1在结直肠癌患者中的表达情况。关键发现：我们建立了放射抗性细胞，其表现出对放射治疗的耐受性和CSC标志物沿着RAP 1的表达升高。RAP 1沉默显示辐射后增强的DNA损伤和减少的CSC标记物的表达。我们观察到RAP 1和KLF 4蛋白之间的强物理相互作用。此外，在CRC患者的肿瘤中观察到较高的RAP 1表达。数据集分析还显示RAP 1高表达与不良预后相关。
Aims: Radiotherapy is predominantly used as one of the treatment modalities to treat local tumor in colorectal cancer (CRC). Hindrance in disease treatment can be attributed to radio-tolerance of cancer stem cells (CSCs) subsistence in the tumor. Understanding the radio-resistant property of CSCs might help in the accomplishment of targeted radiotherapy treatment and increased disease-free survival. Telomeric RAP1 contributes in modulation of various transcription factors leading to aberrant cell proliferation and tumor cell migration. Therefore, we investigated the role of RAP1 in maintaining resistance phenotype and acquired stemness in radio-resistant cells. Main methods: Characterization of HCT116 derived radio-resistant cell (HCT116RR) was performed by cell survival and DNA damage profiling. RAP1 silenced cells were investigated for DNA damage and expression of CSC markers through western blotting and Real-time PCR post-irradiation. Molecular docking and co-immunoprecipitation study were performed to investigate RAP1 and KLF4 interaction followed by RAP1 protein status profiling in CRC patient. Key findings: We established radio-resistant cells, which showed tolerance to radiotherapy and elevated expression of CSC markers along with RAP1. RAP1 silencing showed enhanced DNA damage and reduced expression of CSC markers post-irradiation. We observed strong physical interaction between RAP1 and KLF4 protein. Furthermore, higher RAP1 expression was observed in the tumor of CRC patients. Dataset analysis also revealed that high expression of RAP1 expression is associated with poor prognosis.