抑制SPON2可抑制三阴性乳腺癌的生长。
Knockdown of SPON2 inhibits the growth of triple-negative breast cancer.
作者信息
Hu Xueyi, Su Caiwu, Wei Jian
机构信息
Sinopharm Dongfeng General Hospital, Hubei University of Medicine, Shiyan, Hubei, China.
出版信息
Front Oncol. 2023 Mar 6;13:1141417. doi: 10.3389/fonc.2023.1141417. eCollection 2023.
OBJECTIVE
Spondin-2 (SPON2) is highly expressed in a variety of tumors and has been associated with poor prognosis, but the relationship to triple-negative breast cancer (TNBC) is unclear. The aim of this study is to investigate the expression of SPON2 in TNBC and its function.
METHODS
Immunohistochemistry was used to detect the expression of the SPON2 protein in TNBC and in normal tissue adjacent to cancer and breast fibroadenoma. The GEO database GSE76275 dataset was used to study the expression of SPON2 mRNA in TNBC and non-TNBC. The expression of SPON2 mRNA was detected by qPCR in TNBC cells MDA-MB-231, non-TNBC breast cancer cells MCF-7, and normal breast cells MCF-10A. Kaplan Meier-Plotter database was used to analyze the relationship between SPON2 expression and TNBC prognosis. ShRNA lentivirus was used to knock down high expression of SPON2 in TNBC cells. The effects of knockdown of SPON2 expression on the proliferation, migration, invasion, apoptosis, and subcutaneous tumorigenic ability of TNBC cells in nude mice were analyzed using CCK8, clone formation assay, scratch assay, transwell migration assay, transwell invasion assay, Hoechst apoptosis assay, and tumorigenic ability in nude mice. Transcriptome sequencing of TNBC cells with knockdown SPON2 expression. In combination with the GEO database, GO and KEGG analyses were performed, and psychophysiological interaction Protein-Protein Interaction Networks (PPI) analysis was performed for transcriptome sequencing of the differentially expressed genes. The changes in the expression of PI3K-ATK pathway proteins after SPON2 knockdown were detected by Western blot.
RESULTS
Our study shows that upregulation of SPON2 in TNBC is associated with poorer patient outcomes. Knockdown of SPON2 inhibited TNBC cell proliferation, clone formation, migration, invasion, and tumorigenic ability and promoted apoptosis. Knockdown of SPON2 up-regulated TNBC cell adhesion and down-regulated PI3K-ATK pathway, and PPI results showed that CCL2 was the key protein.
CONCLUSIONS
SPON2 may be a valuable biomarker for the diagnosis and prognosis of TNBC and is a potential therapeutic target for TNBC.
目的
Spondin-2（SPON2）在多种肿瘤中高表达，且与预后不良相关，但与三阴性乳腺癌（TNBC）的关系尚不清楚。本研究旨在探讨SPON2在TNBC中的表达及其功能。
方法
采用免疫组织化学法检测SPON2蛋白在TNBC、癌旁正常组织及乳腺纤维腺瘤中的表达。利用GEO数据库GSE76275数据集研究SPON2 mRNA在TNBC和非TNBC中的表达。采用qPCR检测TNBC细胞MDA-MB-231、非TNBC乳腺癌细胞MCF-7和正常乳腺细胞MCF-10A中SPON2 mRNA的表达。利用Kaplan Meier-Plotter数据库分析SPON2表达与TNBC预后的关系。采用shRNA慢病毒敲低TNBC细胞中SPON2的高表达。运用CCK8、克隆形成实验、划痕实验、transwell迁移实验、transwell侵袭实验、Hoechst凋亡实验及裸鼠致瘤能力实验，分析敲低SPON2表达对TNBC细胞增殖、迁移、侵袭、凋亡及裸鼠皮下致瘤能力的影响。对敲低SPON2表达的TNBC细胞进行转录组测序。结合GEO数据库，进行GO和KEGG分析，并对差异表达基因的转录组测序进行心理生理相互作用蛋白质-蛋白质相互作用网络（PPI）分析。通过蛋白质印迹法检测SPON2敲低后PI3K-ATK通路蛋白表达的变化。
结果
我们的研究表明，TNBC中SPON2的上调与患者较差的预后相关。敲低SPON2可抑制TNBC细胞增殖、克隆形成、迁移、侵袭及致瘤能力，并促进凋亡。敲低SPON2可上调TNBC细胞黏附并下调PI3K-ATK通路，PPI结果显示CCL2是关键蛋白。
结论
SPON2可能是TNBC诊断和预后的有价值生物标志物，也是TNBC的潜在治疗靶点。
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