Objective: To investigate the effects of lncRNA HOTAIR on the proliferation, invasion and migration of lymphoma cells through target gene miR-20a-5p and its molecular mechanism.
Methods: After synthesizing HOTAIR siRNA and siRNA NC plasmids, they were transfected into lymphoma Raji cells, respectively. The expression of HOTAIR mRNA was detected by RT-qPCR. The proliferation, invasion and migration of lymphoma Raji cells were detected by CCK-8 assay, Transwell assay and cell scratch healing assay, respectively. The target gene of lncRNA HOTAIR was predicted by miRcode software, and the relationship between HOTAIR and target gene was analyzed by dual luciferase assay. After synthesis of miR-20a-5p inhibitor and inhibitor NC, Raji cells were transiently transfected. The expression of miR-20a-5p was detected by RT-qPCR, and the effects of down-regulation of miR-20a-5p on the proliferation, invasion and migration of Raji cells were analyzed. The overexpression plasmid of lncRNA HOTAIR and miR-20a-5p mimics were transfected into Raji cells simultaneously to analyze the proliferation, invasion and migration ability of Raji cells. After overexpression or down-regulation of miR-20a-5p, the expression of JAK/STAT3 signaling pathway related proteins was analyzed.
Results: HOTAIR expression in Raji cells was decreased after transfection of HOTAIR siRNA (P<0.01), and miR-20a-5p expression was also decreased after transfection of miR-20a-5p inhibitor (P<0.01). HOTAIR had a targeting and negative regulation relationship with miR-20a-5p (r=-0.826). Silencing HOTAIR promoted the expression of miR-20a-5p and inhibited the proliferation, invasion and migration of Raji cells. Down-regulation of miR-20a-5p expression promoted the proliferation, invasion and migration of Raji cells. Effect of HOTAIR overexpression on the proliferation, invasion and migration of Raji cells could be reversed by up-regulation of miR-20a-5p. Down-regulation of miR-20a-5p expression activated the intracellular JAK/STAT3 signaling pathway.
Conclusion: HOTAIR affects the proliferation, invasion and migration of lymphoma cells by targeting miR-20a-5p, and its mechanism may be related to the activation of JAK/STAT3 signaling pathway.
题目: LncRNA HOTAIR通过靶基因miR-20a-5p调控淋巴瘤细胞 增殖、侵袭、迁移的分子机制研究.
目的: 探讨lncRNA HOTAIR通过靶基因miR-20a-5p对淋巴瘤细胞增殖、侵袭和迁移的影响及其分子机制研究.
方法: 合成HOTAIR siRNA和siRNA NC质粒后，分别转染淋巴瘤Raji细胞，RT-qPCR检测HOTAIR的mRNA表达，分别通过CCK-8实验、Transwell和细胞划痕愈合实验检测Raji细胞的增殖、侵袭和迁移情况。miRcode软件预测lncRNA HOTAIR的靶基因，并通过双荧光素酶实验分析HOTAIR与靶基因的关系。合成miR-20a-5p inhibitor和inhibitor NC后，瞬时转染Raji细胞，RT-qPCR检测miR-20a-5p表达，分析下调miR-20a-5p对Raji细胞的增殖、侵袭和迁移的影响。用lncRNA HOTAIR过表达质粒和miR-20a-5p模拟物同时转染Raji细胞，分析Raji细胞的增殖、侵袭和迁移能力。过表达或下调miR-20a-5p后，分析JAK/STAT3信号通路相关蛋白的表达.
结果: 转染HOTAIR siRNA后Raji细胞中HOTAIR表达降低（P<0.01），转染miR-20a-5p inhibitor后Raji细胞中miR-20a-5p表达降低（P< 0.01）；HOTAIR与miR-20a-5p存在靶向及负调控关系（r=-0.826）；沉默HOTAIR促进miR-20a-5p表达，抑制Raji细胞增殖、侵袭和迁移；下调miR-20a-5p表达促进Raji细胞增殖、侵袭和迁移。上调miR-20a-5p可逆转过表达HOTAIR 对Raji细胞增殖、侵袭和迁移的促进作用。下调miR-20a-5p表达激活了细胞内JAK/STAT3信号通路.
结论: HOTAIR 通过靶向miR-20a-5p影响淋巴瘤细胞的增殖、侵袭和迁移，其作用机制可能与JAK/STAT3信号通路激活有关.
Keywords: HOTAIR; invasion; lymphoma; miR-20a-5p; migration; proliferation.