层粘连蛋白52链的过表达与喉鳞状细胞癌的肿瘤细胞增殖、侵袭及不良预后相关。

Overexpression of Laminin 52 Chain Correlates with Tumor Cell Proliferation, Invasion, and Poor Prognosis in Laryngeal Squamous Cell Carcinoma.

作者信息

Yin Chengyi, Ma Bingliang, Zhang Xilin, Lan Longjiang, Ren Gang, Xu Jue, Wang Jianqiu

机构信息

Department of Otolaryngology, The First Affiliated Hospital, Huzhou University, The First People's Hospital of Huzhou, Huzhou City, Zhejiang Province, China.

Research Department, The First Affiliated Hospital, Huzhou University, The First People's Hospital of Huzhou, Huzhou City, Zhejiang Province, China.

出版信息

J Oncol. 2022 Oct 15;2022:7248064. doi: 10.1155/2022/7248064. eCollection 2022.

DOI:10.1155/2022/7248064

PMID:36284634

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9588344/

Abstract

OBJECTIVE

Laryngeal squamous cell carcinoma (LSCC) is a common malignant tumor. Laminin 52 chain (LAMC2) was reported to be associated with tumorigenesis. This study explored the role of LAMC2 on LSCC progression by regulating the integrin1/FAK/Src/AKT pathway.

METHODS

The level of LAMC2 in 46 LSCC patients was detected by qRT-PCR and western blot. Then the relationship between LAMC2 expression and LSCC malignancy as well as prognosis was analyzed, and the effect of LAMC2 expression on LSCC patient survival was also analyzed using the Kaplan-Meier survival curves. Afterwards, the LSCC cells were transfected with LAMC2 overexpression and knockdown vectors, the effect of LAMC2 on LSCC cell viability, proliferation ability, cell cycle, cell migration, and invasion were detected by CCK-8, colony formation, flow cytometry, wound healing, and Transwell assays. The expression of EMT-related biomarkers and integrin 1/FAK/Src/AKT signaling-related proteins was detected by western blot. Moreover, the effect of LAMC2 on LSCC tumor growth was evaluated by xenograft experiments and western blot.

RESULTS

LAMC2 was expressed at high level in LSCC tissues and associated with poor prognosis. LAMC2 overexpression increased TU177 cell viability, proliferation ability, promoted cell cycle, cell migration, and invasion capacity. The expression of N-cadherin, vimentin, and integrin1/FAK/Src/AKT related proteins was increased, while the expression of E-cadherin protein was decreased. When the LAMC2 knockdown in AMC-HN-8 cells had opposite effects. Furthermore, shLAMC2 decreased tumor volume and the expression of LAMC2, Ki-67 and integrin1, but increased the expression of E-cadherin in LSCC tumor-bearing mice.

CONCLUSION

The findings suggested that LAMC2 was overexpressed in LSCC and correlated with poor prognosis. LAMC2 knockdown inhibited LSCC progression by regulating the integrin1/FAK/Src/AKT signaling pathway. Therefore, LAMC2 could be a target for LSCC therapy.

摘要

目的

喉鳞状细胞癌（LSCC）是一种常见的恶性肿瘤。据报道，层粘连蛋白52链（LAMC2）与肿瘤发生有关。本研究通过调节整合素1/黏着斑激酶（FAK）/Src/蛋白激酶B（AKT）信号通路，探讨LAMC2在LSCC进展中的作用。

方法

采用实时定量聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法检测46例LSCC患者的LAMC2水平。分析LAMC2表达与LSCC恶性程度及预后的关系，并采用Kaplan-Meier生存曲线分析LAMC2表达对LSCC患者生存的影响。随后，用LAMC2过表达和敲低载体转染LSCC细胞，通过细胞计数试剂盒-8（CCK-8）、集落形成、流式细胞术、伤口愈合和Transwell实验检测LAMC2对LSCC细胞活力、增殖能力、细胞周期、细胞迁移和侵袭的影响。采用蛋白质免疫印迹法检测上皮-间质转化（EMT）相关生物标志物和整合素1/FAK/Src/AKT信号相关蛋白的表达。此外，通过异种移植实验和蛋白质免疫印迹法评估LAMC2对LSCC肿瘤生长的影响。

结果

LAMC2在LSCC组织中高表达，且与预后不良相关。LAMC2过表达增加了TU177细胞活力、增殖能力，促进了细胞周期、细胞迁移和侵袭能力。N-钙黏蛋白、波形蛋白和整合素1/FAK/Src/AKT相关蛋白的表达增加，而E-钙黏蛋白的表达降低。当AMC-HN-8细胞中的LAMC2被敲低时，产生相反的效果。此外，在荷瘤小鼠中，短发夹RNA-LAMC2（shLAMC2）降低了肿瘤体积以及LAMC2、Ki-67和整合素1的表达，但增加了E-钙黏蛋白的表达。