LARS1 knockdown suppresses the biological activities of thyroid cancer cells by stimulating autophagy

IF 2.5 4区生物学 Q1 ANATOMY & MORPHOLOGY

Tissue & cell Pub Date : 2025-08-06 DOI:10.1016/j.tice.2025.103075

Congwen Jin, Chuanhong Li, Linlin Chen, Hao Liu, Yousheng Yu, Benxin Chen

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引用次数: 0

Abstract

Background

Given the elevated prevalence of thyroid cancer in China and its status as the leading malignant endocrine tumor, identifying molecular targets for intervention is critical. This study sought to define the functional significance of LARS1 in thyroid cancer pathogenesis and delineate the mechanistic pathways involved.

Materials and methods

We measured LARS1 protein expression in adjacent normal and cancer tissues using immunohistochemistry. First, we knocked down LARS1 in CAL-62 and 8305 C cell lines using si-LARS1 and then inhibited autophagy using an mTOR agonist. Second, we measured cell proliferation, apoptosis rate, invasion, migration and ultrastructure using CCK-8 assay, EdU assay, flow cytometry, TUNEL staining, Transwell assay, wound healing assay and transmission electron microscopy, LC3B protein levels were evaluated by IF staining. ATG7, beclin1, P62 and TIM23 gene expression levels were measured by RT-qPCR; and LC3, ATG7, beclin1, P62 and TIM23 protein expression levels were assessed by western blot.

Results

Elevated LARS1 protein expression was consistently observed in tumor specimens relative to normal counterparts (P < 0.001). Depletion of LARS1 expression resulted in a pronounced attenuation of cellular proliferation concurrent with a substantial elevation in apoptotic rates. Furthermore, the capacities of cancer cells for invasion and migration were markedly impaired following LARS1 knockdown (P < 0.001). This manipulation also led to significant enhancements in the expression levels of both the protein and mRNA encoding ATG7 and beclin1, while diminishing the levels of TIM23 and P62. A concomitant increase in the LC3-II to LC3-I ratio was also documented. Notably, the tumor-suppressive outcomes elicited by si-LARS1 were counteracted upon administration of an mTOR agonist.

Conclusion

LARS1 knockdown suppressed thyroid cancer cell abilities by regulating autophagy activation through mTOR inhibition in vitro.

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LARS1敲低通过刺激自噬抑制甲状腺癌细胞的生物活性

鉴于甲状腺癌在中国的高发病率及其作为主要恶性内分泌肿瘤的地位，确定干预的分子靶点至关重要。本研究旨在确定LARS1在甲状腺癌发病机制中的功能意义，并描述其机制途径。材料与方法采用免疫组化方法检测LARS1蛋白在癌旁和正常组织中的表达。首先，我们使用si-LARS1敲除CAL-62和8305 C细胞系中的LARS1，然后使用mTOR激动剂抑制自噬。其次，采用CCK-8法、EdU法、流式细胞术、TUNEL染色、Transwell染色、创面愈合法和透射电镜检测细胞增殖、凋亡率、侵袭、迁移和超微结构，IF染色检测LC3B蛋白水平。RT-qPCR检测ATG7、beclin1、P62、TIM23基因表达水平；western blot检测LC3、ATG7、beclin1、P62、TIM23蛋白表达水平。结果LARS1蛋白在肿瘤标本中的表达与正常标本一致升高（P <； 0.001）。LARS1表达的缺失导致细胞增殖的明显衰减，同时凋亡率显著升高。此外，LARS1敲低后，癌细胞的侵袭和迁移能力明显受损（P <； 0.001）。这种操作还导致编码ATG7和beclin1的蛋白和mRNA的表达水平显著增强，同时降低了TIM23和P62的表达水平。LC3-II与LC3-I比值的增加也有文献记载。值得注意的是，si-LARS1引起的肿瘤抑制结果在使用mTOR激动剂后被抵消。结论lars1基因敲低可通过mTOR抑制甲状腺癌细胞自噬激活，从而抑制甲状腺癌细胞的能力。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Tissue & cell 医学-解剖学与形态学

CiteScore

3.90

自引率

0.00%

发文量

234

期刊介绍： Tissue and Cell is devoted to original research on the organization of cells, subcellular and extracellular components at all levels, including the grouping and interrelations of cells in tissues and organs. The journal encourages submission of ultrastructural studies that provide novel insights into structure, function and physiology of cells and tissues, in health and disease. Bioengineering and stem cells studies focused on the description of morphological and/or histological data are also welcomed. Studies investigating the effect of compounds and/or substances on structure of cells and tissues are generally outside the scope of this journal. For consideration, studies should contain a clear rationale on the use of (a) given substance(s), have a compelling morphological and structural focus and present novel incremental findings from previous literature.