CRYAB基因对结直肠癌干细胞样特性的影响及其机制。

Effects of the CRYAB gene on stem cell-like properties of colorectal cancer and its mechanism.

作者信息

Dai Ang, Guo Xiaohong, Yang Xiaoqing, Li Min, Fu Yanxin, Sun Qing

机构信息

College of Basic Medicine, Weifang Medical University, Weifang, Shandong; Department of Pathology, The First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital, Shandong medicine and Health Key Laboratory of Clinical Pathology, Shandong Lung Cancer Institute, Shandong Institute of Nephrology, Jinan, Shandong, China.

Department of Pathology, The First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital, Shandong medicine and Health Key Laboratory of Clinical Pathology, Shandong Lung Cancer Institute, Shandong Institute of Nephrology, Jinan, Shandong, China.

出版信息

J Cancer Res Ther. 2022 Sep;18(5):1328-1337. doi: 10.4103/jcrt.jcrt_212_22.

DOI:10.4103/jcrt.jcrt_212_22

PMID:36204880

Abstract

AIMS

Alpha B-crystallin (CRYAB), a known molecular chaperone, is involved in the occurrence and development of various tumor types. However, the function of CRYAB in colorectal cancer stem cells (CSCs) remains unknown. This study aimed to elucidate the role and possible regulatory mechanisms of CRYAB in the cancer stem cell-like phenotype of colorectal cancer (CRC).

SUBJECTS AND METHODS

The expression of CRYAB in patients with CRC and lymph node metastasis at various stages and its relationship with overall survival were detected using the TCGA database. In this study, CRC-CSCs were enriched from HCT116 and Caco2 cells with serum-free suspension culture. The CRYAB gene, stemness-related genes, and mesenchymal markers were detected via quantitative real-time PCR (qRT-PCR) in CRC cells. Then, CRYAB-HCT116S and CRYAB-Caco2S cell lines were established by lentivirus-mediated overexpression of CRYAB. Self-renewal ability and stemness features were measured by the sphere formation assay and flow cytometry. The tumorigenesis capacity in vivo was inspected in nude mice. The functions of CRYAB on CSC proliferation, migration, and invasion were examined using colony formation and the transwell assay. Finally, the Wnt/β-catenin pathway-related mRNAs and proteins were detected via qRT-PCR and western blotting.

RESULTS

The expression of CRYAB in CRC is related to the clinical phase and prognosis, except with lymphoid metastasis. CRYAB expression was elevated in CSCs. Upregulation of CRYAB enhanced the expression of CSC-related genes and mesenchymal markers. The capacity to form colonospheres, tumorigenesis, cell proliferation, and metastasis were significantly advanced in CRYAB-overexpressed cells. Moreover, CRYAB dramatically suppressed β-catenin degradation and downregulated the expression of p-GSK-3β.

CONCLUSIONS

CRYAB maintains CSC formation via the Wnt/β-catenin pathway in CRCs, which may, therefore, function as vital molecular targets for CRC therapeutic strategies.

摘要

目的

αB晶状体蛋白（CRYAB）是一种已知的分子伴侣，参与多种肿瘤类型的发生和发展。然而，CRYAB在结直肠癌干细胞（CSCs）中的功能尚不清楚。本研究旨在阐明CRYAB在结直肠癌（CRC）癌干细胞样表型中的作用及可能的调控机制。

对象与方法

利用TCGA数据库检测CRYAB在不同阶段CRC患者及有淋巴结转移患者中的表达及其与总生存期的关系。在本研究中，采用无血清悬浮培养从HCT116和Caco2细胞中富集CRC-CSCs。通过定量实时PCR（qRT-PCR）检测CRC细胞中CRYAB基因、干性相关基因和间充质标志物。然后，通过慢病毒介导的CRYAB过表达建立CRYAB-HCT116S和CRYAB-Caco2S细胞系。通过成球试验和流式细胞术测量自我更新能力和干性特征。在裸鼠中检测体内成瘤能力。使用集落形成试验和Transwell试验检测CRYAB对CSC增殖、迁移和侵袭的作用。最后，通过qRT-PCR和蛋白质印迹法检测Wnt/β-连环蛋白通路相关的mRNA和蛋白质。

结果

CRYAB在CRC中的表达与临床分期和预后相关，但与淋巴转移无关。CRYAB在CSCs中的表达升高。CRYAB的上调增强了CSC相关基因和间充质标志物的表达。CRYAB过表达的细胞中形成结肠球的能力、成瘤能力、细胞增殖和转移能力均显著提高。此外，CRYAB显著抑制β-连环蛋白的降解并下调p-GSK-3β的表达。