Data_Sheet_1_LncRNA DHRS4-AS1 Inhibits the Stemness of NSCLC Cells by Sponging miR-224-3p and Upregulating TP53 and TET1.PDF

Non-small cell lung cancer (NSCLC) is the leading cause of cancer-related death. This study aimed to examine the roles of DHRS4-AS1/miR-224-3p signaling in the cancer cell stemness of NSCLC. Real-time PCR showed that DHRS4-AS1 was downregulated in cancerous tissues, and bioinformatics analysis revealed that high DHRS4-AS1 expression indicated a good prognosis for NSCLC patients. Sphere and colony formation assays showed that DHRS4-AS1 overexpression significantly suppressed NSCLC cell colony formation and stem cell-like properties. DHRS4-AS1 also abrogated the expression of OCT4, SOX2, CD34, and CD133, markedly inhibited the expression of epithelial-mesenchymal transition (EMT)-related factors, N-cadherin, ZEB1, and Vimentin, and increased E-cadherin expression in spheres. Furthermore, luciferase reporter assays and real-time PCR analysis demonstrated that DHRS4-AS1 and miR-224-3p were antagonistically repressed in NSCLC cells. RNA immunoprecipitation (RIP) analysis revealed that DHRS4-AS1 interacted with miR-224-3p. DHRS4-AS1 partially reversed the miR-224-3p-decreased TP53 and TET1, resulting in the inhibition of tumor growth in vivo. Finally, TP53 and TET1 were antagonistically regulated by DHRS4-AS1 and miR-224-3p in NSCLC cells. In conclusion, TP53- and TET1-associated DHRS4-AS1/miR-224-3p axis is an essential mechanism by which NSCLC modulates cancer cell stemness.

非小细胞肺癌（NSCLC）是导致癌症相关死亡的首要原因。本研究旨在探讨DHRS4-AS1/miR-224-3p信号通路在NSCLC癌细胞干性中的作用。实时PCR检测显示，DHRS4-AS1在癌组织中呈现下调表达，生物信息学分析揭示，高表达DHRS4-AS1预示NSCLC患者的良好预后。球体和集落形成实验表明，DHRS4-AS1过表达显著抑制NSCLC细胞的集落形成和干细胞样特性。DHRS4-AS1还消除了OCT4、SOX2、CD34和CD133的表达，显著抑制了上皮间质转化（EMT）相关因子N-钙粘蛋白、ZEB1和波形蛋白的表达，并增加了球体中E-钙粘蛋白的表达。此外，荧光素酶报告基因实验和实时PCR分析证实，DHRS4-AS1和miR-224-3p在NSCLC细胞中呈现拮抗性抑制。RNA免疫沉淀（RIP）分析发现，DHRS4-AS1与miR-224-3p相互作用。DHRS4-AS1部分逆转了miR-224-3p下调的TP53和TET1，从而在体内抑制肿瘤生长。最终，TP53和TET1在NSCLC细胞中受到DHRS4-AS1和miR-224-3p的拮抗性调控。综上所述，TP53-和TET1相关的DHRS4-AS1/miR-224-3p轴是NSCLC调节癌细胞干性的关键机制（Non-small cell lung cancer (NSCLC), DHRS4-AS1/miR-224-3p signaling, cancer cell stemness, real-time PCR, bioinformatics analysis, sphere and colony formation assays, OCT4, SOX2, CD34, CD133, epithelial-mesenchymal transition (EMT), N-cadherin, ZEB1, Vimentin, E-cadherin, luciferase reporter assays, RNA immunoprecipitation (RIP), TP53, TET1）。