长链非编码RNA DRAIC通过抑制miR-223-3p促进肺癌细胞增殖和转移

lncRNA DRAIC promotes proliferation and metastasis of lung cancer cells by inhibiting miR-223-3p

目的探究长链非编码RNA(lncRNA)DRAIC对miR-223-3p的调节作用及对肺癌细胞增殖和转移的影响。方法选取2019年1月至2020年6月河北工程大学附属医院收治的90例非小细胞肺癌患者的癌组织和癌旁组织,分析癌症基因组图谱(TCGA)数据中肺癌组织lncRNA DRAIC的表达及其与患者预后生存期的关系,实时定量聚合酶链反应(RT-qPCR)检测肺癌组织lncRNA DRAIC和miR-223-3p的表达水平并进行Pearson相关性分析;Starbase数据库预测及双荧光素酶实验验证lncRNA DRAIC与miR-223-3p的靶向关系;将经慢病毒包装的lncRNA DRAIC和miR-223-3p慢病毒质粒分别转染至肺癌细胞A549和H520中,分为对照组、si-NC组、si-DRAIC组、pcDNA组、pcDNA-DRAIC组、DRAIC+miR-NC组、DRAIC+miR-223-3p组;MTT法和集落形成实验、划痕实验和Transwell小室实验分别检测细胞增殖能力、迁移能力和侵袭能力;RT-qPCR和蛋白印迹法检测JAK2和STAT3的mRNA和蛋白的表达;裸鼠成瘤实验检测移植瘤的发展。结果TCGA数据显示肺癌组织lncRNA DRAIC的表达水平高于癌旁组织,且预后生存期与lncRNA DRAIC的表达水平呈负相关;与癌旁组织相比,肺癌组织中lncRNA DRAIC水平升高,miR-223-3p水平降低,且lncRNA DRAIC与miR-223-3p呈负相关;lncRNA DRAIC水平与肿瘤大小、TNM分期、淋巴结转移、分化程度呈正相关(P<0.05)。lncRNA DRAIC与miR-223-3p存在结合位点,lncRNA DRAIC过表达抑制miR-223-3p的表达(P<0.05);lncRNA DRAIC过表达能够上调JAK2和STAT3的mRNA和蛋白水平,促进细胞增殖、划痕愈合和增加侵袭细胞数,促进移植瘤生长(P<0.05)。另外,miR-223-3p可部分逆转lncRNA DRAIC对肺癌细胞的恶性表型。结论lncRNA DRAIC通过抑制miR-223-3p促进肺癌细胞的增殖、迁移、侵袭及体内移植瘤的发展,且可能通过激活JAK2/STAT3信号通路发挥作用。

Objective To explore the regulatory effect of lncRNA DRAIC on mi R-223-3p and its effect on proliferation and metastasis of lung cancer cells. Method The expression of lncRNA DRAIC in lung cancer tissue and paracancer tissues, and its relationship with prognosis and survival time of 90 patients with non-small cell lung cancer were analyzed by TCGA data from January 2019 to June 2020 in Affiliated Hospital of Hebei University of Engineering. The expression of lncRNA DRAIC and mi R-223-3p in lung cancer tissue was detected by real-time quantitative polymerase chain reaction( RT-q PCR) and the correlation of Pearson was analyzed. Starbase prediction and double luciferase test were used to verify the targeting relationship of lncRNA DRAIC to mi R-223-3p. The lentivirus-packaged lncRNA DRAIC and mi R-223-3p lentivirus plasmids were transfected into lung cancer cells A549 and H520 respectively, and were divided into control group, si-NC group, si-DRAIC group, pc DNA group, pc DNA-DRAIC group, DRAIC+mi R-NC group and DRAIC+mi R-223-3p group. The ability of cell proliferation, migration and invasion were detected by MTT assay and colony formation test, scratch test and Transwell chamber test, respectively. RT-q PCR and western blot method were used to detect the expression of m RNA and protein of JAK2 and STAT3. The tumorigenesis experiment in nude mice was used to detect the development of transplanted tumors. Result TCGA data showed that the expression level of lncRNA DRAIC in lung cancer tissues was higher than that in paracancer tissues, and the prognostic survival time was negatively correlated with the expression level of lncRNA DRAIC. Compared with the paracancer tissues, the level of lncRNA DRAIC in lung cancer tissues increased, and the level of mi R-223-3p decreased, and the levels of lncRNA DRAIC and mi R-223-3p showed a negative correlation. The level of lncRNA DRAIC is positively correlated with tumor size, TNM stage, lymph node metastasis, and differentiation(P<0. 05). There is a binding site between lncRNA DRAIC and mi R-223-3p, and the overexpression of lncRNA DRAIC inhibits the expression of mi R-223-3p(P<0. 05) and can upregulate the levels of m RNA and protein of JAK2 and STAT3, promote cell proliferation, increase scratch healing and the number of invaded cells, and promote the growth of transplanted tumors(P<0. 05). And mi R-223-3p can partially reverse the malignant phenotype of lncRNA DRAIC on lung cancer cells. Conclusion lncRNA DRAIC promotes the proliferation, migration and invasion of lung cancer cells and the development of transplanted tumors in vivo by inhibiting mi R-223-3p, and it may play a role by activating the JAK2/STAT3 signaling pathway.