长链非编码 RNA LINC00520 通过下调 EGFR 使 PI3K/Akt 信号通路失活，从而阻止皮肤鳞状细胞癌的进展。
Long noncoding RNA LINC00520 prevents the progression of cutaneous squamous cell carcinoma through the inactivation of the PI3K/Akt signaling pathway by downregulating EGFR.
机构信息
Department of Dermatology, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China.
出版信息
Chin Med J (Engl). 2019 Feb;132(4):454-465. doi: 10.1097/CM9.0000000000000070.
BACKGROUND
Long noncoding RNAs (lncRNAs) play pivotal roles in various malignant tumors. Epidermal growth factor receptor (EGFR) signaling is associated with the pathogenesis of cutaneous squamous cell carcinoma (cSCC). This study aimed to explore the role of LINC00520 in the development of cSCC via EGFR and phosphoinositide 3-kinase-protein kinase B (PI3K/Akt) signaling pathways.
METHODS
A microarray analysis was applied to screen differentially expressed lncRNAs in cSCC samples. The A431 cSCC cell line was transfected and assigned different groups. The expression patterns of LINC00520, EGFR, and intermediates in the PI3K/Akt pathway were characterized using reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blotting analysis. Cell proliferation, migration, and invasion were detected using the MTT assay, scratch test, and Transwell assay, respectively. Cell-based experiments and a tumorigenicity assay were conducted to assess the effect of LINC00520 on cSCC progression. This study was ended in September 2017. Comparisons between two groups were analyzed with t-test and comparisons among multiple groups were analyzed using one-way analysis of variance. The nonparametric Wilcoxon rank sum test was used to analyze skewed data. The enumerated data were analyzed using the chi-square test or Fisher exact test.
RESULTS
Data from chip GSE66359 revealed depletion of LINC00520 in cSCC. Cells transfected with LINC00520 vector and LINC00520 vector + si-EGFR showed elevated LINC00520 level but decreased levels of the EGFR, PI3K, AKT, VEGF, MMP-2 and MMP-9 mRNAs and proteins, and inhibition of the growth, migration and adhesion of cSCC cells, while the si-LINC00520 group showed opposite trends (all P < 0.05). Compared with the LINC00520 vector group, the LINC00520 vector + si-EGFR group showed decreased levels of the EGFR, PI3K, AKT, VEGF, MMP-2 and MMP-9 mRNAs and proteins, and inhibition of the growth, migration and adhesion of cSCC cells, while the LINC00520 vector + EGFR vector group showed opposite results (all P < 0.05).
CONCLUSION
Based on our results, LINC00520-targeted EGFR inhibition might result in the inactivation of the PI3K/Akt pathway, thus inhibiting cSCC development.
背景
长链非编码 RNA（lncRNA）在各种恶性肿瘤中发挥着关键作用。表皮生长因子受体（EGFR）信号与皮肤鳞状细胞癌（cSCC）的发病机制有关。本研究旨在通过 EGFR 和磷脂酰肌醇 3-激酶-蛋白激酶 B（PI3K/Akt）信号通路探索 LINC00520 在 cSCC 发展中的作用。
方法
应用微阵列分析筛选 cSCC 样本中差异表达的 lncRNA。转染 A431 cSCC 细胞系并分配不同的组。采用逆转录定量聚合酶链反应（RT-qPCR）和 Western blot 分析检测 LINC00520、EGFR 及 PI3K/Akt 通路中间产物的表达模式。分别采用 MTT 测定、划痕试验和 Transwell 测定检测细胞增殖、迁移和侵袭。通过细胞实验和肿瘤发生实验评估 LINC00520 对 cSCC 进展的影响。本研究于 2017 年 9 月结束。采用 t 检验比较两组间差异，采用单因素方差分析比较多组间差异。采用非参数 Wilcoxon 秩和检验分析偏态数据。采用卡方检验或 Fisher 确切检验分析计数数据。
结果
芯片 GSE66359 数据显示 cSCC 中 LINC00520 耗竭。转染 LINC00520 载体和 LINC00520 载体+si-EGFR 的细胞 LINC00520 水平升高，但 EGFR、PI3K、AKT、VEGF、MMP-2 和 MMP-9 mRNA 和蛋白水平降低，cSCC 细胞的生长、迁移和黏附受到抑制，而 si-LINC00520 组则呈现相反趋势（均 P<0.05）。与 LINC00520 载体组相比，LINC00520 载体+si-EGFR 组 cSCC 细胞的 EGFR、PI3K、AKT、VEGF、MMP-2 和 MMP-9 mRNA 和蛋白水平降低，生长、迁移和黏附受到抑制，而 LINC00520 载体+EGFR 载体组则呈现相反结果（均 P<0.05）。
结论
根据我们的结果，LINC00520 靶向 EGFR 抑制可能导致 PI3K/Akt 通路失活，从而抑制 cSCC 发展。
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