Department of Oncology, Shanxian Central Hospital, Heze, China.
Department of Obstetrics, Jining Medical College Affiliated Hospital, Jining, China.
J Biochem Mol Toxicol. 2021 Jun;35(6):1-11. doi: 10.1002/jbt.22749. Epub 2021 Feb 25.
This paper investigates the function of lncRNA DARS-AS1 in cervical cancer (CC) as well as its in-depth mechanism. The differential expression of DARS-AS1 and ATP1B2 were analyzed based on The Cancer Genome Atlas and the Genotype-Tissue Expression databases, and the survival rate was measured using Kaplan-Meier survival analysis. Biological function experiments were performed to detect cell proliferation, invasion, and migration. Quantitative real-time polymerase chain reaction was carried out to detect the expression of DARS-AS1 and ATP1B2. Western blot analysis was utilized to assess the protein levels of ATP1B2 and cGMP-PKG pathway-related proteins. DARS-AS1 was expressed at high levels in CC tissues and cell lines, and high expression of DARS-AS1 indicated a lower survival rate. CCK-8 and colony formation assays revealed that the overexpression of DARS-AS1 promoted the proliferation of CC cells. Furthermore, bioinformatics analysis suggested that the cGMP-PKG pathway ranks as the first pathway enriched by the differential genes that correlated with DARS-AS1 (|r| > 0.4). ATP1B2, as a cGMP-PKG pathway-related gene, was significantly correlated with the overall survival of CC patients. We further confirmed that ATP1B2 was lowly expressed in CC and negatively correlated with the DARS-AS1 expression. Then, biological function experiments exhibited that the promotion of cell proliferation, invasion, and migration resulted due to the upregulation of DARS-AS1 could be canceled by ATP1B2 overexpression. Finally, Western blot revealed that upregulation of DARS-AS1 could activate the cGMP-PKG pathway, while overexpression of ATP1B2 reversed this activation. Our study revealed that DARS-AS1/ATP1B2 contributes to regulating the progression of CC at least partially by modulating the cGMP-PKG pathway.
本文研究了长链非编码 RNA DARS-AS1 在宫颈癌 (CC) 中的功能及其深入的机制。基于癌症基因组图谱和基因型组织表达数据库分析了 DARS-AS1 和 ATP1B2 的差异表达，并通过 Kaplan-Meier 生存分析测量了生存率。进行了生物功能实验以检测细胞增殖、侵袭和迁移。通过实时定量聚合酶链反应检测 DARS-AS1 和 ATP1B2 的表达。利用 Western blot 分析评估 ATP1B2 和 cGMP-PKG 通路相关蛋白的蛋白水平。DARS-AS1 在 CC 组织和细胞系中表达水平较高，DARS-AS1 高表达提示生存率较低。CCK-8 和集落形成实验表明，DARS-AS1 的过表达促进了 CC 细胞的增殖。此外，生物信息学分析表明，cGMP-PKG 通路是与 DARS-AS1 相关的差异基因（|r|>0.4）富集的第一个通路。ATP1B2 作为 cGMP-PKG 通路相关基因，与 CC 患者的总生存率显著相关。我们进一步证实，ATP1B2 在 CC 中表达水平较低，与 DARS-AS1 的表达呈负相关。然后，生物学功能实验表明，由于 DARS-AS1 的上调导致的细胞增殖、侵袭和迁移的促进作用可以被 ATP1B2 的过表达所取消。最后，Western blot 显示，DARS-AS1 的上调可以激活 cGMP-PKG 通路，而过表达 ATP1B2 则逆转了这种激活。我们的研究表明，DARS-AS1/ATP1B2 通过调节 cGMP-PKG 通路至少部分参与调控 CC 的进展。
