SPINK5 inhibits esophageal squamous cell carcinoma metastasis via immune activity

IF 3.2 4区医学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Journal of Gene Medicine Pub Date : 2024-03-05 DOI:10.1002/jgm.3667

Jie Chen, Juncheng Lu, Zhiqiang Chen, Zihao Liu, Yuejun Sun, Shuyan He, Yedong Mi, Yi Gao, Dong Shen, Qingfeng Lin

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Abstract

Background

Esophageal squamous cell carcinoma (ESCC) is a predominant subtype of esophageal cancer with relatively high mortality worldwide. Serine peptidase inhibitor Kazal-type 5 (SPINK5) is reported to be downregulated in ESCC. However, its explicit role in ESCC remains further investigation.

Methods

The tumor tissues and adjacent non-cancerous tissues were obtained from 196 patients with ESCC for the determination of SPINK5 mRNA levels. Additionally, the relationship between SPINK5 mRNA levels and clinicopathological features of ESCC patients was explored. The effects of SPINK5 on the invasion and migration of ESCC cells were assessed using Transwell assays. Furthermore, SPINK5 mRNA and LEKTI protein were measured in ESCC cell lines after treatment with poly (I:C), lipopolysaccharide (LPS) or unmethylated CpG DNA. Moreover, the correlation between expression of SPINK5 and nuclear factor-kappa B (NF-κB) signaling pathway-related genes was analyzed in the TCGA-ESCC cohort, and the effects of SPINK5 on NF-κB transcription was analyzed using a luciferase reporter gene assay. Finally, the correlations between SPINK5 and infiltration of immune cells, immune scores, stromal scores and ESTIMATE (i.e., Estimation of STromal and Immune cells in MAlignant Tumor tissues using Expression data) scores were explored.

Results

SPINK5 mRNA levels were downregulated in tumor tissues, which was significantly correlated with higher lymph node metastases. Overexpressed SPINK5 inhibited cell invasion and migration in ESCC cell lines. Mechanistically, LPS-induced activation of Toll-like receptor 4 (TLR4) decreased SPINK5 mRNA and LEKTI in KYSE150 and KYSE70 cells. Spearman correlation analysis revealed that SPINK5 mRNA was significantly negatively correlated with a total of seven NF-κB signaling pathway-related genes in TCGA-ESCC patients. Moreover, downregulation of SPINK5 increased and upregulation of SPINK5 decreased the activity of the NF-κB promoter in HEK293T cells. Finally, immune cells infiltration analysis revealed that SPINK5 was significantly correlated with the infiltration of various immune cells, stromal scores, immune scores and ESTIMATE scores.

Conclusions

SPINK5 plays critical roles in the TLR4/NF-κB pathway and immune cells infiltration, which might contribute to the ESCC metastasis. The findings of the present study may provide a promising biomarker for the diagnosis and treatment of esophageal squamous cell carcinoma.

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SPINK5通过免疫活性抑制食管鳞状细胞癌转移

背景：食管鳞状细胞癌（ESCC）是食管癌的主要亚型，在全球范围内死亡率相对较高。据报道，丝氨酸肽酶抑制剂卡扎尔 5 型（SPINK5）在 ESCC 中被下调。然而，其在 ESCC 中的明确作用仍有待进一步研究：方法：从 196 例 ESCC 患者的肿瘤组织和邻近的非癌组织中获取 SPINK5 mRNA 水平。此外，还探讨了 SPINK5 mRNA 水平与 ESCC 患者临床病理特征之间的关系。使用Transwell试验评估了SPINK5对ESCC细胞侵袭和迁移的影响。此外，还测定了ESCC细胞系经多聚（I：C）、脂多糖（LPS）或未甲基化CpG DNA处理后的SPINK5 mRNA和LEKTI蛋白。此外，在TCGA-ESCC队列中分析了SPINK5与核因子-kappa B（NF-κB）信号通路相关基因表达的相关性，并使用荧光素酶报告基因实验分析了SPINK5对NF-κB转录的影响。最后，还探讨了SPINK5与免疫细胞浸润、免疫评分、基质评分和ESTIMATE（即利用表达数据估算恶性肿瘤组织中的基质和免疫细胞）评分之间的相关性：结果：SPINK5 mRNA水平在肿瘤组织中下调，这与淋巴结转移率的升高显著相关。过表达的 SPINK5 可抑制 ESCC 细胞系的细胞侵袭和迁移。从机理上讲，LPS诱导的Toll样受体4（TLR4）激活降低了KYSE150和KYSE70细胞中的SPINK5 mRNA和LEKTI。斯皮尔曼相关分析表明，在TCGA-ESCC患者中，SPINK5 mRNA与7个NF-κB信号通路相关基因呈显著负相关。此外，在 HEK293T 细胞中，SPINK5 的下调会增加 NF-κB 启动子的活性，而 SPINK5 的上调则会降低 NF-κB 启动子的活性。最后，免疫细胞浸润分析表明，SPINK5与各种免疫细胞的浸润、基质评分、免疫评分和ESTIMATE评分显著相关：结论：SPINK5在TLR4/NF-κB通路和免疫细胞浸润中起着关键作用，可能会导致ESCC转移。本研究的结果可能为食管鳞状细胞癌的诊断和治疗提供了一种有前景的生物标记物。

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

Journal of Gene Medicine 医学-生物工程与应用微生物

CiteScore

6.40

自引率

0.00%

发文量

审稿时长

6-12 weeks

期刊介绍： The aims and scope of The Journal of Gene Medicine include cutting-edge science of gene transfer and its applications in gene and cell therapy, genome editing with precision nucleases, epigenetic modifications of host genome by small molecules, siRNA, microRNA and other noncoding RNAs as therapeutic gene-modulating agents or targets, biomarkers for precision medicine, and gene-based prognostic/diagnostic studies. Key areas of interest are the design of novel synthetic and viral vectors, novel therapeutic nucleic acids such as mRNA, modified microRNAs and siRNAs, antagomirs, aptamers, antisense and exon-skipping agents, refined genome editing tools using nucleic acid /protein combinations, physically or biologically targeted delivery and gene modulation, ex vivo or in vivo pharmacological studies including animal models, and human clinical trials. Papers presenting research into the mechanisms underlying transfer and action of gene medicines, the application of the new technologies for stem cell modification or nucleic acid based vaccines, the identification of new genetic or epigenetic variations as biomarkers to direct precision medicine, and the preclinical/clinical development of gene/expression signatures indicative of diagnosis or predictive of prognosis are also encouraged.