BCAT1 Overexpression Promotes Proliferation, Invasion, and Wnt Signaling in Non-Small Cell Lung Cancers
BCAT1 Overexpression Promotes Proliferation, Invasion, and Wnt Signaling in Non-Small Cell Lung Cancers
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BCAT1 过表达促进非小细胞肺癌的增殖、侵袭和 Wnt 信号转导
DOI：
10.2147/ott.s237306
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发表时间：
2020-01-01
影响因子：
4
通讯作者：
Dong, Qianze
中科院分区：
文献类型：
--
作者：
Lin, Xiumin;Tan, Shutao;Dong, Qianze
简介 BCAT1 失调与癌发生有关。然而，其在人类非小细胞肺癌（NSCLC）中的临床意义和生物学作用尚不清楚。方法采用免疫组化方法检测107例肺癌组织中BCAT1蛋白的表达。使用 MTT、集落形成测定、Matrigel 侵袭测定、Western blot、RNA 测序和荧光素酶报告基因测定检查 BCAT1 的生物学作用和潜在机制。结果我们发现 BCAT1 在 107 个肺癌组织中的 60 个中表达上调，并与淋巴结转移、晚期和较短的总生存期相关。癌症基因组图谱 (TCGA) 和 ONCOMINE 数据分析也表明 BCAT1 在人类 NSCLC 组织中升高。 BCAT1蛋白在肺癌细胞系中高于正常支气管上皮细胞系。 BCAT1 过表达增加了 BEAS-2B 和 H1299 细胞系的细胞生长速率、集落数量和侵袭能力，而 BCAT1 siRNA 降低了细胞增殖速率、集落数量并抑制了侵袭。 RNA 测序 (RNA-seq) 和基因集富集分析 (GSEA) 分析表明 BCAT1 过表达激活了 Wnt/Myc 信号传导。蛋白质印迹显示，BCAT1 增加了 BEAS-2B 和 H1299 细胞系中 MMP7、细胞周期蛋白 D1、c-Myc 的蛋白表达，并减少了 E-钙粘蛋白和 p27 的表达。进一步的实验表明，BCAT1 过表达会提高 Wnt 报告荧光素酶活性并增加激活 β-catenin 蛋白，同时下调 p-β-catenin 蛋白表达。 BCAT1 敲除显示出相反的效果。 TCGA 数据分析表明 BCAT1 与 c-Myc、细胞周期蛋白 D1 和 MMP7 mRNA 之间呈正相关。使用抑制剂 (ICG-001) 阻断 Wnt 信号传导可下调 c-Myc、细胞周期蛋白 D1、MMP7 的表达，并消除 BCAT1 对这些蛋白质的上调作用。结论 总之，我们的数据表明 BCAT1 在人类 NSCLC 中过度表达。 BCAT1 可能通过调节 Wnt 信号通路促进细胞增殖和侵袭。
Introduction Dysregulation of BCAT1 has been implicated in carcinogenesis. However, its clinical significance and biological roles in human non-small cell lung cancer (NSCLC) are not clear. Methods Immunohistochemistry was used to examine the protein expression of BCAT1 in 107 cases of lung cancer tissues. Biological roles and potential mechanisms of BCAT1 were examined using MTT, colony formation assay, Matrigel invasion assay, Western blot, RNA-sequencing, and luciferase reporter assay. Results We found BCAT1 was upregulated in 60 of 107 lung cancer tissues and correlated with nodal metastasis, advanced stages and short overall survival. The Cancer Genome Atlas (TCGA) and ONCOMINE data analyses also indicated that BCAT1 was elevated in human NSCLC tissues. BCAT1 protein was higher in lung cancer cell lines than in normal bronchial epithelial cell line. BCAT1 overexpression increased the cell growth rate, colony numbers and invasion abilities in both BEAS-2B and H1299 cell lines, while BCAT1 siRNA decreased the cell proliferation rate, colony numbers, and inhibited invasion. RNA-sequencing (RNA-seq) and Gene Set Enrichment Analysis (GSEA) analyses indicated that BCAT1 overexpression activated Wnt/Myc signaling. Western blot revealed that BCAT1 increased protein expression of MMP7, cyclin D1, c-Myc, and decreased E-cadherin and p27 in the BEAS-2B and H1299 cell lines. Further experiments showed that BCAT1 overexpression elevated Wnt reporter luciferase activity and increased activate β-catenin protein while downregulating p-β-catenin protein expression. BCAT1 knockdown showed the opposite effects. TCGA data analysis suggested positive correlations between BCAT1 and c-Myc, cyclin D1, and MMP7 mRNA. Blockage of Wnt signaling using an inhibitor (ICG-001) downregulated c-Myc, cyclin D1, MMP7 expressions and abolished the upregulating effects of BCAT1 on these proteins. Conclusion In summary, our data showed that BCAT1 was overexpressed in human NSCLCs. BCAT1 facilitated cell proliferation and invasion possibly through regulation of the Wnt signaling pathway.