Bai Yue, Lang Lili, Zhao Wentao, Niu Rong
Department One of Thoracic Surgery, Gansu Provincial Cancer Hospital, Gansu, People's Republic of China.
Department of Radiology, Gansu Provincial Cancer Hospital, Gansu, People's Republic of China.
Onco Targets Ther. 2019 Dec 17;12:11195-11206. doi: 10.2147/OTT.S198367. eCollection 2019.
Our present study aimed to further investigate the molecular basis of long non-coding RNA homeobox A11 antisense (HOXA11-AS) in the tumorigenesis of non-small cell lung cancer (NSCLC).
HOXA11-AS, microRNA-148a-3p (miR-148a-3p), and DNA methyltransferase 1 (DNMT1) mRNA levels were measured by RT-qPCR assay. DNMT1 protein level was determined by Western blot assay. Cell proliferative capacity and apoptotic rate were determined by CCK-8 assay and flow cytometry analysis, respectively. The relationships of HOXA11-AS, miR-148a-3p, and DNMT1 were tested through bioinformatics analysis, luciferase assay, and RNA pull down assay. Mouse xenograft models of NSCLC were established to examine the biological function of HOXA11-AS in vivo.
HOXA11-AS expression was notably upregulated and miR-148a-3p expression was conspicuously downregulated in NSCLC tissues and cells. HOXA11-AS knockdown curbed NSCLC cell proliferation and promoted cell apoptosis through directly increasing miR-148a-3p expression. Moreover, miR-148a-3p overexpression suppressed NSCLC cell proliferation and induced cell apoptosis. HOXA11-AS functioned as a competing endogenous RNA (ceRNA) of miR-148a-3p to increase DNMT1 expression in NSCLC cells. And, DNMT1 upregulation weakened the influence of HOXA11-AS1 loss on NSCLC cell proliferation and apoptosis. Additionally, HOXA11-AS knockdown suppressed NSCLC xenograft growth by upregulating miR-148a-3p and downregulating DNMT1 in vivo.
HOXA11-AS facilitated NSCLC tumorigenesis through miR-148a-3p/DNMT1 axis in vitro and in vivo, deepening our understanding of the molecular basis of HOXA11-AS in the development of NSCLC.
本研究旨在进一步探究长链非编码RNA同源盒A11反义链（HOXA11-AS）在非小细胞肺癌（NSCLC）肿瘤发生中的分子基础。
采用RT-qPCR法检测HOXA11-AS、微小RNA-148a-3p（miR-148a-3p）和DNA甲基转移酶1（DNMT1）的mRNA水平。通过蛋白质印迹法测定DNMT1蛋白水平。分别采用CCK-8法和流式细胞术分析检测细胞增殖能力和凋亡率。通过生物信息学分析、荧光素酶报告基因检测和RNA下拉实验检测HOXA11-AS、miR-148a-3p和DNMT1之间的关系。建立NSCLC小鼠异种移植模型以在体内检测HOXA11-AS的生物学功能。
HOXA11-AS在NSCLC组织和细胞中的表达显著上调，而miR-148a-3p的表达明显下调。敲低HOXA11-AS可抑制NSCLC细胞增殖并通过直接增加miR-148a-3p的表达促进细胞凋亡。此外，miR-148a-3p过表达可抑制NSCLC细胞增殖并诱导细胞凋亡。HOXA11-AS作为miR-148a-3p的竞争性内源RNA（ceRNA）增加NSCLC细胞中DNMT1的表达。并且，DNMT1上调减弱了HOXA11-AS1缺失对NSCLC细胞增殖和凋亡的影响。此外，敲低HOXA11-AS可通过上调miR-148a-3p和下调DNMT1抑制NSCLC异种移植瘤在体内的生长。
HOXA11-AS在体外和体内通过miR-148a-3p/DNMT1轴促进NSCLC的肿瘤发生，加深了我们对HOXA11-AS在NSCLC发生发展中分子基础的理解。
