Molecule interacting with CasL-like protein 2 (MICALL2) is believed to regulate cytoskeleton dynamics, tight junction formation, and neurite outgrowth. However, its biological role and the underlying mechanism in colorectal cancer (CRC) remain largely elusive. qRT-PCR, Western blotting and immunohistochemistry assays were used to detect the expression levels of different genes. Next, mass spectrometry, co-immunoprecipitation and immunofluorescence staining were used to detect the interactions of proteins. Furthermore, MTT assay, colony formation assay, wound-healing assays and xenograft tumor models were performed to demonstrate the functions of MICALL2 in CRC. In addition, transcriptome sequencing and Western blotting were conducted to verify the mechanism of MICALL2 in CRC. We found that both mRNA and protein levels of MICALL2 are up-regulated in colorectal cancer tissues compared with non-tumor tissues and that its overexpression is closely correlated with poor prognosis. Ubiquitin E3 ligase Tripartite motif-containing protein 21 (TRIM21) mediated MICALL2 ubiquitination and proteasome-dependent degradation, negatively correlated with MICALL2 levels, and reversely regulated the tumorigenic activity of MICALL2 in CRC. Functional studies confirmed that MICALL2 promoted colorectal cancer cell growth and migration via the Wnt/β-catenin signaling pathway. As a substrate of ubiquitinase TRIM21, MICALL2 enhances the growth and migration of colorectal cancer cells and activates the Wnt/β-catenin signaling pathway.

中文翻译：

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MICALL2 作为泛素化酶 TRIM21 的底物调节结直肠癌的肿瘤发生

与 CasL 样蛋白 2 (MICALL2) 相互作用的分子被认为可以调节细胞骨架动力学、紧密连接形成和神经突生长。然而，其在结直肠癌 (CRC) 中的生物学作用和潜在机制在很大程度上仍不清楚。qRT-PCR、Western blotting 和免疫组织化学分析用于检测不同基因的表达水平。接下来，使用质谱法、免疫共沉淀和免疫荧光染色来检测蛋白质的相互作用。此外，进行了 MTT 测定、集落形成测定、伤口愈合测定和异种移植肿瘤模型以证明 MICALL2 在 CRC 中的功能。此外，还进行了转录组测序和Western blotting以验证MICALL2在CRC中的作用机制。我们发现与非肿瘤组织相比，结直肠癌组织中 MICALL2 的 mRNA 和蛋白水平均上调，其过度表达与不良预后密切相关。泛素 E3 连接酶含三联基序的蛋白 21 (TRIM21) 介导 MICALL2 泛素化和蛋白酶体依赖性降解，与 MICALL2 水平呈负相关，并反向调节 MICALL2 在 CRC 中的致瘤活性。功能研究证实 MICALL2 通过 Wnt/β-catenin 信号通路促进结直肠癌细胞的生长和迁移。作为泛素化酶 TRIM21 的底物，MICALL2 促进结直肠癌细胞的生长和迁移，并激活 Wnt/β-catenin 信号通路。与 MICALL2 水平呈负相关，并反向调节 MICALL2 在 CRC 中的致瘤活性。功能研究证实 MICALL2 通过 Wnt/β-catenin 信号通路促进结直肠癌细胞的生长和迁移。作为泛素化酶 TRIM21 的底物，MICALL2 促进结直肠癌细胞的生长和迁移，并激活 Wnt/β-catenin 信号通路。与 MICALL2 水平呈负相关，并反向调节 MICALL2 在 CRC 中的致瘤活性。功能研究证实 MICALL2 通过 Wnt/β-catenin 信号通路促进结直肠癌细胞的生长和迁移。作为泛素化酶 TRIM21 的底物，MICALL2 促进结直肠癌细胞的生长和迁移，并激活 Wnt/β-catenin 信号通路。