摘要
目的探讨壳多糖酶3样蛋白1(CHI3L1)在大肠癌细胞辐射抵抗中的作用和调控机制。方法从TCGA数据库下载获得380例大肠癌患者的CHI3L1 mRNA表达数据,包括380例大肠癌组织样本和51例癌旁组织样本,其中31例为配对组织样本。此外,保存于宁波大学实验室的40例配对大肠癌及其癌旁组织样本亦用于本研究。运用TCGA数据库和免疫组化法分析CHI3L1在大肠癌组织与癌旁组织中的差异表达情况。在不同剂量辐射后,检测CHI3L1过表达对大肠癌细胞克隆形成能力的影响。通过免疫荧光实验、细胞凋亡实验、细胞周期实验检测辐射后CHI3L1表达对大肠癌细胞增殖及凋亡的影响。通过蛋白质印迹实验探索CHI3L1调控大肠癌细胞辐射抵抗的分子机制。结果CHI3L1在大肠癌组织中呈高表达。不同剂量辐射后,相对于野生型大肠癌细胞,CHI3L1过表达组大肠癌细胞克隆形成比例增加,细胞坏死比例减少;CHI3L1过表达组大肠癌细胞增殖增加,G2/M期细胞占比减少。大肠癌细胞中过表达CHI3L1伴随着Wnt4表达的显著上调辐射可诱导CHI3L1和Wnt4表达上调,且CHI3L1过表达细胞Wnt4表达上调的幅度更显著;回复实验表明,在相同剂量辐射下,Wnt4敲除组细胞克隆形成比例明显减少。结论CHI3L1过表达可以减少辐射引起的大肠癌细胞坏死,减弱辐射导致的增殖抑制和G2/M期阻滞,并且通过上调Wnt4表达引起大肠癌细胞辐射抵抗。
Objective To investigate the role and regulatory mechanism of Chitinase-3-like protein 1(CHI3L1)in radiation resistance of colorectal cancer(CRC)cells.Methods CHI3L1 differential expression in CRC samples and adjacent normal samples was analyzed based on the TCGA database.A colony formation assay was performed to evaluate the colony forming ability after CHI3L1 overexpression in CRC cells.Immunofluorescence,cell apoptosis and cell cycle assays were used to assess the difference in radiation effects between normal and overexpression CRC cells.Western blot assays were conducted to explore the molecular mechanism of CHI3L1 in the radiation resistance of CRC cells.Results CHI3L1 was highly expressed in CRC tissues.The colony numbers of the CHI3L1-overexpressing CRC cell group were significantly higher than those of the control group after radiation.CHI3L1-overexpressing cell lines showed substantially less nuclear necrosis after radiation.Furthermore,the CHI3L1-overexpressing group had a higher proportion of proliferative cells and a lower proportion of G2/M phase cells.Moreover,overexpression of CHI3L1 was accompanied by Wnt4 upregulation in CRC cells.Radiation could induce upregulation of CHI3L1 and Wnt4,which was more pronounced in the CHI3L1-overexpressing groups.The rescue experiment showed that the colony numbers significantly decreased in the Wnt4 knockdown groups.Conclusions Overexpression of CHI3L1 reduces radiation-induced necrosis and inhibited proliferation of CRC cells,as well as radiation-induced G2/M phase arrest.CHI3L1 induces radiation resistance by upregulating Wnt4 expression in CRC cells.
作者
金铭
叶爽
郑真
刘开泰
JIN Ming;YE Shuang;ZHENG Zhen;LIU Kaitai(Ningbo Medical Center Lihuili Hospital Ningbo 315040,Zhejiang,China)
出处
《现代实用医学》
2023年第4期441-446,共6页
Modern Practical Medicine
基金
浙江省基础公益研究计划项目(LY21H160013)
宁波市自然科学基金(202003N4203)
浙江省医药卫生科技计划项目(2020KY861)。
