长链非编码RNA HOTAIR通过靶基因调控淋巴瘤细胞增殖、侵袭和迁移的分子机制
[The Molecular Mechanism of LncRNA HOTAIR Regulating the Proliferation, Invasion and Migration of Lymphoma Cells through Target Gene ].
作者信息
Lei A-Ming, Zhu Ping-An, Fu Li-Mei, Liao Xin, Azi Gu-Li, Zhu Ping
机构信息
Department of Hematology, The First People's Hospital of Chenzhou City, Chenzhou 423000, Hunan Province, China.
Department of Hematology, The First People's Hospital of Chenzhou City, Chenzhou 423000, Hunan Province, China.E-mail:
出版信息
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2023 Feb;31(1):89-95. doi: 10.19746/j.cnki.issn.1009-2137.2023.01.014.
OBJECTIVE
To investigate the effects of lncRNA on the proliferation, invasion and migration of lymphoma cells through target gene and its molecular mechanism.
METHODS
After synthesizing siRNA and siRNA NC plasmids, they were transfected into lymphoma Raji cells, respectively. The expression of 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 was predicted by miRcode software, and the relationship between and target gene was analyzed by dual luciferase assay. After synthesis of inhibitor and inhibitor NC, Raji cells were transiently transfected. The expression of was detected by RT-qPCR, and the effects of down-regulation of on the proliferation, invasion and migration of Raji cells were analyzed. The overexpression plasmid of lncRNA and mimics were transfected into Raji cells simultaneously to analyze the proliferation, invasion and migration ability of Raji cells. After overexpression or down-regulation of , the expression of JAK/STAT3 signaling pathway related proteins was analyzed.
RESULTS
expression in Raji cells was decreased after transfection of siRNA (<0.01), and expression was also decreased after transfection of inhibitor (<0.01). had a targeting and negative regulation relationship with (r=-0.826). Silencing promoted the expression of and inhibited the proliferation, invasion and migration of Raji cells. Down-regulation of expression promoted the proliferation, invasion and migration of Raji cells. Effect of overexpression on the proliferation, invasion and migration of Raji cells could be reversed by up-regulation of . Down-regulation of expression activated the intracellular JAK/STAT3 signaling pathway.
CONCLUSION
affects the proliferation, invasion and migration of lymphoma cells by targeting , and its mechanism may be related to the activation of JAK/STAT3 signaling pathway.
目的
通过靶基因研究长链非编码RNA（lncRNA）对淋巴瘤细胞增殖、侵袭和迁移的影响及其分子机制。
方法
合成小干扰RNA（siRNA）和siRNA阴性对照（NC）质粒后，分别转染至淋巴瘤Raji细胞。采用逆转录定量聚合酶链反应（RT-qPCR）检测mRNA表达。分别采用细胞计数试剂盒-8（CCK-8）法、Transwell法和细胞划痕愈合试验检测淋巴瘤Raji细胞的增殖、侵袭和迁移能力。用miRcode软件预测lncRNA的靶基因，通过双荧光素酶试验分析lncRNA与靶基因的关系。合成lncRNA抑制剂和抑制剂阴性对照后，对Raji细胞进行瞬时转染。采用RT-qPCR检测lncRNA表达，分析lncRNA下调对Raji细胞增殖、侵袭和迁移的影响。将lncRNA过表达质粒和lncRNA模拟物同时转染至Raji细胞，分析Raji细胞的增殖、侵袭和迁移能力。lncRNA过表达或下调后，分析Janus激酶/信号转导子和转录激活子3（JAK/STAT3）信号通路相关蛋白的表达。
结果
转染lncRNA siRNA后，Raji细胞中lncRNA表达降低（P<0.01），转染lncRNA抑制剂后lncRNA表达也降低（P<0.01）。lncRNA与靶基因存在靶向负调控关系（r=-0.826）。沉默lncRNA促进靶基因表达，抑制Raji细胞的增殖、侵袭和迁移。lncRNA表达下调促进Raji细胞的增殖、侵袭和迁移。lncRNA过表达对Raji细胞增殖、侵袭和迁移的影响可被靶基因上调所逆转。lncRNA表达下调激活细胞内JAK/STAT3信号通路。
结论
lncRNA通过靶向靶基因影响淋巴瘤细胞的增殖、侵袭和迁移，其机制可能与激活JAK/STAT3信号通路有关。
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