Zhao Qi, Zhang Kexin, Li Zugui, Zhang Hao, Fu Fangmei, Fu Junjie, Zheng Minying, Zhang Shiwu
Department of Pathology, Tianjin Union Medical Center, Nankai University, Tianjin, China.
Tianjin Medical University, Tianjin, China.
Front Cell Dev Biol. 2021 Jul 16;9:696871. doi: 10.3389/fcell.2021.696871. eCollection 2021.
Our previous studies have confirmed that cobalt chloride (CoCl) or chemoradiotherapy could induce the formation of polyploid tumor giant cells (PGCCs). Polyploid giant cancer cells are a special subpopulation of cancer cells that contribute to solid tumor heterogeneity. The size of PGCC was at least three times larger than regular diploid cancer cells. PGCCs have the properties of cancer stem cells (CSCs) and can express CSC markers CD44 and CD133. Daughter cells derived from PGCCs have strong proliferation, infiltration and migration abilities. However, the detailed molecular mechanism of daughter cells expressing mesenchymal phenotype and displaying strong abilities of proliferation and migration is unclear. As a plasminogen receptor, S100A10 which is closely associated with the invasion and metastasis of malignant tumors, was highly expressed in PGCCs with their daughter cells. In this study, CoCl was used to induce the formation of PGCCs in LoVo and HCT116 CRC cells. Cell functional experiments, co-immunoprecipitation, MG132 and ginkgolic acid treatment, western blot, and ChIP-Seq were used to identify the mechanism of S100A10 nuclear location. The proliferation and migration abilities of PGCCs and their daughter cells decreased significantly after S100A10 knockdown. In the control cells, S100A10 was mainly ubiquitinated, while in PGCCs and daughter cells, S100A10 was mainly SUMOylated, which was associated with S100A10 nuclear location. After SUMO1 was inhibited, the nuclear S100A10 in PGCCs and daughter cells decreased, and their proliferation and migration abilities significantly decreased. ChIP-Seq combined with real-time fluorescent quantitative PCR showed that S100A10 regulated the expression of neutrophil defensin 3 (), receptor-type tyrosine-protein phosphatase N2 (), and rho guanine nucleotide exchange factor 18 (), which were associated with actin dynamics and cytoskeleton remodeling. The expression of S100A10 in the nuclei and cytoplasm of rectal cancer after neoadjuvant chemoradiation (nCRT) and liver metastases increased compared with that in rectal cancer without nCRT. Taken together, the expression and nuclear localization of S100A10 modified by SUMOylation were associated with the high proliferation and migration of PGCCs and their daughter cells, and the differentiation, metastases, and relapse of CRCs by regulating the expression of , , and .
我们之前的研究已证实，氯化钴（CoCl）或放化疗可诱导多倍体肿瘤巨细胞（PGCCs）形成。多倍体巨癌细胞是癌细胞的一个特殊亚群，它导致实体瘤异质性。PGCC的大小至少是正常二倍体癌细胞的三倍。PGCC具有癌症干细胞（CSCs）的特性，可表达CSC标志物CD44和CD133。源自PGCC的子代细胞具有很强的增殖、浸润和迁移能力。然而，子代细胞表达间充质表型并展现出强大增殖和迁移能力的详细分子机制尚不清楚。作为一种纤溶酶原受体，与恶性肿瘤侵袭和转移密切相关的S100A10在PGCC及其子代细胞中高表达。在本研究中，用CoCl诱导LoVo和HCT116结肠直肠癌（CRC）细胞形成PGCC。采用细胞功能实验、免疫共沉淀、MG132和银杏酸处理、蛋白质免疫印迹法以及染色质免疫沉淀测序（ChIP-Seq）来确定S100A10核定位的机制。S100A10敲低后，PGCC及其子代细胞的增殖和迁移能力显著下降。在对照细胞中，S100A10主要被泛素化，而在PGCC及其子代细胞中，S100A10主要被小泛素样修饰物（SUMO）化，这与S100A10核定位有关。SUMO1被抑制后，PGCC及其子代细胞中的核S100A10减少，其增殖和迁移能力显著下降。ChIP-Seq结合实时荧光定量PCR表明，S100A10调节中性粒细胞防御素3、受体型酪氨酸蛋白磷酸酶N2和rho鸟嘌呤核苷酸交换因子18的表达，这些因子与肌动蛋白动力学和细胞骨架重塑有关。新辅助放化疗（nCRT）及肝转移后直肠癌细胞核和细胞质中S100A10的表达相较于未进行nCRT的直肠癌有所增加。综上所述，经SUMO化修饰的S100A10的表达及核定位与PGCC及其子代细胞的高增殖和迁移有关，并通过调节相关基因的表达与CRC的分化、转移及复发有关。
