lncRNA作为竞争性内源RNA在非小细胞肺癌中的作用

长链非编码RNA(long non-coding RNA,lncRNA)参与肿瘤的多种生理、病理进程.研究表明,lncRNA可通过与微小RNA (microRNA, mi RNA)反应元件相互作用,并与其他RNA分子形成竞争性内源RNA (competing endogenous RNA,ceRNA)的调控网络,参与基因的表达调控.lncRNA以ceRNA方式参与非小细胞肺癌(non-small cell lung cancer,NSCLC)的发生发展过程,为揭示NSCLC的分子机理开拓了新的思路,也为NSCLC的治疗提供新的靶点.本文在课题组前期发现NSCLC相关ceRNA基础上,主要讨论lncRNA作为ceRNA在NSCLC中高表达、低表达及治疗相关方面的作用.     

Knockdown of lncRNA ZFAS1-suppressed non–small cell lung cancer progression via targeting the miR-150-5p/HMGA2 signaling

Non–small cell lung cancer (NSCLC) is the main type of lung malignancy. Early diagnosis and treatments for NSCLC are far from satisfactory due to the limited knowledge of the molecular mechanisms regarding NSCLC progression. Long noncoding RNA (lncRNA) ZNFX1 antisense RNA1 (ZFAS1) has been implicated for its functional role in the progression of malignant tumors. This study aimed to determine the ZFAS1 expression from lung cancer clinical samples and to explore the molecular mechanisms underlying ZFAS1-modulated NSCLC progression. Experimental assays revealed that clinical samples and cell lines of lung malignant tumors showed an upregulation of ZFSA1. ZFAS1 expression was markedly upregulated in the lung tissues from patients with advanced stage of this malignancy. The loss-of-function assays showed that knockdown of ZFAS1-suppressed NSCLC cell proliferative, as well as invasive potentials, increased NSCLC cell apoptotic rates in vitro and also attenuated tumor growth of NSCLC cells in the nude mice. Further experimental evidence showed that ZFAS1 inversely affected miR-150-5p expression and positively affected high-mobility group AT-hook 2 (HMGA2) expression in NSCLC cell lines. MiR-150-5p inhibition or HMGA2 overexpression counteracted the effects of ZFAS1 knockdown on NSCLC cell proliferative, invasive potentials and apoptotic rates. In light of examining the clinical lung cancer samples, miR-150-5p expression was downregulated and the HMGA2 expression was highly expressed in the lung cancer tissues compared with normal ones; the ZFAS1 expression showed a negative correlation with miR-150-5p expression but a positive correlation with HMGA2 expression in lung cancer tissues. To summarize, we, for the first time, demonstrated the inhibitory effects of ZFAS1 knockdown on NSCLC cell progression, and the results from mechanistic studies indicated that ZFAS1-mediated NSCLC progression cells via targeting miR-150-5p/HMGA2 signaling.     

lncRNA DGCR5在非小细胞肺癌组织中的表达及其与临床病理特征的相关性分析

摘要 目的：探究lncRNA DGCR5在非小细胞肺癌(NSCLC)组织中的表达及其与临床病理特征的相关性。方法：选取2020年1月至2021年12月在我院肿瘤科收治的进行手术治疗的NSCLC患者86例,在手术期间从患者获得肿瘤和非肿瘤的肺癌旁组织样本。采用qRT-PCR测定肿瘤组织及癌旁组织中lncRNA DGCR5表达水平。分析lncRNA DGCR5表达水平与NSCLC患者性别、年龄、临床分期、T分期、N分期等临床病理参数的关系,lncRNA DGCR5表达水平与患者预后总生存期（OS）和无进展生存期（PFS）的关系。结果：与癌旁组织相比,lncRNA DGCR5在NSCLC肿瘤组织中的表达水平相对较低,差异具有统计学意义（P<0.01）。lncRNA DGCR5表达与肿瘤分化程度、TNM分期、肿瘤体积、淋巴转移和远处转移之间存在明显相关性,差异具有统计学意义（P<0.05）。采用Kaplan-Meier法进行生存分析,研究发现lncRNA DGCR5高表达组中位OS及中位DFS分别显著高于lncRNA DGCR5低表达组（P<0.05）。低分化程度、II+ IIIa临床分期、N1-N3淋巴转移、远处转移、及lncRNA DGCR5 低表达均与NSCLC患者总生存率和无进展生存率相关。结论：LncRNA DGCR5在NSCLC患者肿瘤组织中的表达量降低,NSCLC患者血LncRNA DGCR5表达水平与分化程度、TNM分期、淋巴转移、远处转移及预后具有相关性。LncRNA DGCR5可作为早期诊断和治疗NSCLC的新型生物标志物。     

Long non-coding RNA ZFAS1 promotes pancreatic cancer proliferation and metastasis by sponging miR-497-5p to regulate HMGA2 expression

The lncRNA ZFAS1 plays a carcinogenic regulatory role in many human tumours, but it is rarely reported in pancreatic cancer. We identify the role and molecular mechanisms of ZFAS1 in pancreatic cancer. The expression of ZFAS1, miR-497-5p and HMGA2 in pancreatic cancer tissues was detected by qRT-PCR. Pancreatic cancer data in The Cancer Genome Atlas were also included in this study. CCK8, EdU, transwell and scratch wound assays were used to investigate the biological effects of ZFAS1 in pancreatic cancer cells. MS2-RIP, RNA pull-down, RNA-ChIP and luciferase reporter assays were used to clarify the molecular biological mechanisms of ZFAS1 in pancreatic cancer. The role of ZFAS1 in vivo was also confirmed via xenograft experiments. ZFAS1 was overexpressed in pancreatic cancer tissues. ZFAS1 promoted the growth and metastasis of pancreatic cancer cells, and miR-497-5p acted as a tumour suppressor gene in pancreatic cancer by targeting HMGA2. We also demonstrated that ZFAS1 exerts its effects by promoting HMGA2 expression through decoying miR-497-5p. We also found that ZFAS1 promoted the progression of pancreatic cancer in vivo by modulating the miR-497-5p/HMGA2 axis. In conclusion, this study revealed a new role for and the molecular mechanisms of ZFAS1 in pancreatic cancer, identifying ZFAS1 as a novel target for the diagnosis and treatment of pancreatic cancer.Subject terms: Oncogenes, Cell invasion, Long non-coding RNAs     

Long noncoding RNA lnc-sox5 modulates CRC tumorigenesis by unbalancing tumor microenvironment

Long non-coding RNAs (LncRNAs) have been recently regarded as systemic regulators in multiple biologic processes including tumorigenesis. In this study, we observed the expression of lncRNA lnc-sox5 was significantly increased in colorectal cancer (CRC). Despite the CRC cell growth, cell cycle and cell apoptosis was not affected by lnc-sox5 knock-down, lnc-sox5 knock-down suppressed CRC cell migration and invasion. In addition, xenograft animal model suggested that lnc-sox5 knock-down significantly suppressed the CRC tumorigenesis. Our results also showed that the expression of indoleamine 2,3-dioxygenase 1 (IDO1) was significantly reduced by lnc-sox5 knock-down and therefore modulated the infiltration and cytotoxicity of CD3⁺CD8⁺T cells. Taken together, these results suggested that lnc-sox5 unbalances tumor microenvironment to regulate colorectal cancer progression.     

LncRNA RP11-89 facilitates tumorigenesis and ferroptosis resistance through PROM2-activated iron export by sponging miR-129-5p in bladder cancer

Long non-coding RNAs (lncRNAs) act as important regulators of tumorigenesis and development in bladder cancer. However, the underlying molecular mechanisms remain elusive. We previously identified a novel lncRNA signature related to immunity and progression in bladder cancer. Here we further explored the function of RP11-89, a lncRNA discovered in the previous signature. Loss- and gain-of function experiments were performed using CCK-8 assay, flow cytometry, Transwell assays, scratch tests and subcutaneous nude mouse models. High-throughput RNA sequencing was conducted to identify dysregulated genes in bladder cancer cells with RP11-89 knockdown or overexpression. Regulation of RP11-89 on miR-129-5p and PROM2 was explored through luciferase reporter assay, RIP assay and RNA pull-down assay. RP11-89 promoted cell proliferation, migration and tumorigenesis and inhibited cell cycle arrest via the miR-129-5p/PROM2 axis. We found that RP11-89 “sponges” miR-129-5p and upregulates PROM2. Elevated PROM2 in cells was associated with attenuated ferroptosis through iron export, formation of multivesicular bodies and less mitochondrial abnormalities. We demonstrated that RP11-89 is a novel tumorigenic regulator that inhibits ferroptosis via PROM2-activated iron export. RP11-89 may serve as a potential biomarker for targeted therapy in bladder cancer.Subject terms: Tumour biomarkers, Tumour biomarkers     

Long non‐coding RNA 00312 regulated by HOXA5 inhibits tumour proliferation and promotes apoptosis in Non‐small cell lung cancer

Non‐small cell lung cancer (NSCLC) is the most prevalent type of lung cancer. The abnormal expression of many long non‐coding RNAs (lncRNAs) has been reported involved in the progression of various tumours, which can be used as diagnostic indicators or antitumour targets. Here, we found that the long non‐coding RNA 00312 was down‐regulated in paired NSCLC tissues and correlated with poor clinical outcome; decreased linc00312 expression in NSCLC was associated with larger and later stage tumours. Functional experiments showed that linc00312 could inhibit cell proliferation and promote apoptosis in vitro and in vivo. Furthermore, we found that HOXA5 could bind in the promoter of linc00312 and up‐regulated the expression of it. Moreover, linc00312 was down‐regulated in the plasma of NSCLC patients compared with that of healthy volunteers or other pulmonary diseases patients. Taken together, our findings indicated that linc00312 could be a novel diagnosis biomarker and a promising therapeutic target for NSCLC.     

血清外泌体lncRNA PCGEM1、miR-129-5p与非小细胞肺癌患者临床病理特征及预后的关系

摘要 目的：研究血清外泌体长链非编码核糖核酸（lncRNA）前列腺癌基因表达标记1（PCGEM1）、微小核糖核酸（miR）-129-5p与非小细胞肺癌（NSCLC）患者临床病理特征及预后的关系。方法：选取2016年2月-2018年1月南京脑科医院收治的125例NSCLC患者作为NSCLC组,同期选取体检的70例健康人群作为健康组。采集两组静脉血,提取血清外泌体;采用实时定量聚合酶链式反应（qRT-PCR）检测血清外泌体lncRNA PCGEM1、miR-129-5p表达情况;采用Pearson相关性分析lncRNA PCGEM1与miR-129-5p的关系。并分析血清外泌体lncRNA PCGEM1、miR-129-5p与NSCLC患者临床病理特征的关系。对NSCLC患者行5年随访,绘制Kaplan-Meier曲线分析预后情况,多因素Cox比例风险回归模型分析预后不良危险因素,受试者工作特征（ROC）曲线分析lncRNA PCGEM1、miR-129-5p对NSCLC预后的预测价值。结果：：NSCLC组lncRNA PCGEM1相对表达量高于健康组,miR-129-5p相对表达量低于健康组（P＜0.05）。血清外泌体lncRNA PCGEM1相对表达量与miR-129-5p表达呈负相关（r= -0.420,P＜0.05）。血清外泌体lncRNA PCGEM1、miR-129-5p表达与患者TNM分期、分化程度、淋巴结转移有关（P＜0.05）。Kplan-Meier生存曲线显示,lncRNA PCGEM1低表达组5年生存率69.05%高于lncRNA PCGEM1高表达组35.53%,miR-129-5p高表达组5年生存率68.09%高于miR-129-5p低表达组33.80%。多因素Cox比例风险回归显示,TNM分期III期、有淋巴结转移、lncRNA PCGEM1高表达、miR-129-5p低表达为NSCLC患者预后不良的独立危险因素（P＜0.05）。ROC曲线显示,lncRNA PCGEM1、miR-129-5p联合检测对NSCLC预后的预测曲线下面积（AUC）为0.865,预测价值高于两者单独预测。结论：NSCLC患者血清外泌体lncRNA PCGEM1表达上调、miR-129-5p表达下调,二者表达与NSCLC患者TNM分期、分化程度、淋巴结转移有关,且与患者预后密切相关,对NSCLC预后不良具有较好预测价值。     

AMPKα loss promotes KRAS-mediated lung tumorigenesis

AMP-activated protein kinase (AMPK) is a critical sensor of energy status that coordinates cell growth with energy balance. In non-small cell lung cancer (NSCLC) the role of AMPKα is controversial and its contribution to lung carcinogenesis is not well-defined. Furthermore, it remains largely unknown whether long non-coding RNAs (lncRNAs) are involved in the regulation of AMPK-mediated pathways. Here, we found that loss of AMPKα in combination with activation of mutant KRAS^G12D increased lung tumour burden and reduced survival in Kras^LSLG12D/+/AMPKα^fl/fl mice. In agreement, functional in vitro studies revealed that AMPKα silencing increased growth and migration of NSCLC cells. In addition, we identified an AMPKα-modulated lncRNA, KIMAT1 (ENSG00000228709), which in turn regulates AMPKα activation by stabilizing the lactate dehydrogenase B (LDHB). Collectively, our study indicates that AMPKα loss promotes KRAS-mediated lung tumorigenesis and proposes a novel KRAS/KIMAT1/LDHB/AMPKα axis that could be exploited for therapeutic purposes.Subject terms: Cancer models, Non-small-cell lung cancer     

Long non-coding RNA DANCR promotes colorectal tumor growth by binding to lysine acetyltransferase 6A

Recent studies have demonstrated that long non-coding RNAs (lncRNAs) play critical roles in cancer development and progression. However, the mechanism by which lncRNAs contribute to colorectal cancer remains unclear. In this study, we identified the lncRNA, DANCR, which was upregulated in colorectal cancer. The upregulation of DANCR expression was associated with shorter patient survival time. DANCR depletion decreased cell proliferation, cell cycle progression, and tumorigenesis in a subcutaneous mouse xenograft model system. We further demonstrated that DANCR bound with lysine acetyltransferase 6A. This binding was essential for KAT6A acetyltransferase activity and thus, it influenced the expression of KAT6A target genes. Our data indicated that DANCR functions as an oncogenic lncRNA that promotes tumor development and progression. Therefore, DANCR may be a target molecule for colorectal cancer treatment.     

CCAT2 enhances autophagy-related invasion and metastasis via regulating miR-4496 and ELAVL1 in hepatocellular carcinoma

Autophagy is thought to contribute to the pathogenesis of many diseases, including cancer. Long non-coding RNA (lncRNA) CCAT2 functions as an oncogene in a variety of tumours. However, it is still unknown whether CCAT2 is involved in autophagy and metastasis of hepatocellular carcinoma (HCC). In our study, we found that lncRNA CCAT2 expression was significantly increased in HCC tissue and was correlated with advanced stage and venous invasion. Further experiments revealed that CCAT2 induced autophagy and promoted migration and invasion in vitro and in vivo. Mechanistic investigations found that CCAT2 involved in HCC by regulating miR-4496/Atg5 in cytoplasm. In nucleus, CCAT2 bound with ELAVL1/HuR to facilitate HCC progression. Our findings suggest that CCAT2 is an oncogenic factor in the progression of HCC with different regulatory mechanisms and may serve as a target for HCC therapy.