长链非编码RNA H19、HOTTIP预测可切除局部进展期胃癌新辅助化疗疗效的研究

目的研究血浆中长链非编码RNA H19、HOTTIP预测可切除局部进展期胃癌新辅助化疗(NAC)疗效的价值。方法前瞻性纳入2020年8月至2021年5月期间于青岛大学附属烟台毓璜顶医院就诊的40例T3~4aN;M0期胃癌患者和40例胃良性疾病患者,在患者入院后未行任何治疗前检测患者血浆中H19和HOTTIP的表达。胃癌患者采用CAPEOX化疗方案,2个疗程后再次检测血浆中H19和HOTTIP的表达,同时检测其他临床项目并评估胃癌患者NAC治疗的效果,将完全缓解和部分缓解归为客观缓解,完全缓解、部分缓解和疾病稳定归为疾病控制。比较H19和HOTTIP在不同患者中的表达差异,采用受试者操作特征曲线(ROC)评估H19和HOTTIP诊断可切除局部进展期胃癌的价值。结果 40例进展期胃癌患者接受NAC治疗后,T降期13例,T未降期27例;客观缓解25例,疾病控制35例。NAC治疗前,胃癌患者血浆中H19和HOTTIP的中位相对表达量均高于胃良性疾病患者(H19:1.42比0.98,Z=–3.835,P<0.001;HOTTIP:2.15比1.04,Z=–5.062,P<0.001),且二者在T降期和疾病控制患者中均低于T未降期和疾病进展(5例)患者(T降期情况:H19:1.12比1.54,Z=–2.960,P=0.002;HOTTIP:1.49比2.30,Z=–2.310,P=0.019;疗效情况:H19:1.39比2.48,Z=–3.211,P<0.001;HOTTIP:1.96比3.25,Z=–2.393,P=0.014)。NAC治疗后,胃癌患者血浆中H19和HOTTIP的中位相对表达量均低于NAC治疗前(H19:1.12比1.42,Z=–3.965,P<0.001;HOTTIP:1.30比2.15,Z=–4.839,P<0.001),但二者NAC治疗前后的变化量值在T降期和T未降期以及客观控制和疾病进展患者中比较差异均无统计学意义(P>0.05)。H19和HOTTIP无论单独或联合区分胃癌与胃良性疾病的ROC曲线下面积值均>0.7。结论血浆中H19和HOTTIP有可能是胃癌的一种潜在肿瘤标志物,其对胃癌的诊断价值较高,血浆中H19和HOTTIP表达水平较低的胃癌患者可能对NAC治疗较敏感。     

沉默lncRNA HOTTIP通过上调miR-663a表达增加非小细胞肺癌细胞系放射敏感性

目的 探讨lncRNA HOTTIP通过调控miR-663a表达对4个非小细胞肺癌细胞系放射敏感性影响。方法 用0、2、4、6、8 Gy分别照射H838、H157、A549、H1299细胞系,采用克隆形成实验检测细胞存活情况。qRT-PCR检测细胞中HOTTIP和miR-663a表达水平。以A549、H1299细胞为研究对象,沉默HOTTIP表达、过表达miR-663a后用克隆形成实验检测细胞存活情况。流式细胞术检测细胞凋亡情况,Western blot检测Cleaved caspase-3、Cleaved PARP和γ-H2AX表达。双荧光素酶报告基因实验和qRT-PCR检测验证HOTTIP和miR-663a的靶向关系。结果 HOTTIP在放射耐受的H157、A549、H1299细胞中表达上调,miR-663a表达下调。沉默HOTTIP或过表达miR-663a均可抑制A549、H1299细胞存活(放射增敏比分别为1.562、1.507),促进Cleaved caspase-3、Cleaved PARP和γ-H2AX表达,促进放射照射诱导细胞凋亡。miR-663a是HOTTIP的靶基因,HOTTIP可负性调控miR-663a的表达。抑制miR-663a表达可逆转沉默HOTTIP对肺癌细胞系放射敏感性的影响。结论 沉默HOTTIP通过上调miR-663a表达,抑制肺癌细胞系存活,促进其凋亡,从而提高肺癌细胞系的放射敏感性。     

长链非编码RNA HOTTIP 通过调控细胞凋亡水平促进非小细胞肺癌增殖

背景与目的：探索非小细胞肺癌(non-small cell lung cancer,NSCLC)有效的早期诊断及预后标志物具有重要的科学意义和临床价值。长链非编码RNA(lncRNA)在肿瘤中的作用逐渐引起研究者的关注。本研究拟分析lncRNA HOTTIP在NSCLC中的表达情况及对NSCLC细胞增殖水平的影响和机制。方法：采用实时定量PCR(quantitative real-time PCR,qRT-PCR)检测方法检测54例NSCLC组织及其对应癌旁组织的HOTTIP表达情况,分析NSCLC组织HOTTIP表达与临床病理参数的相关性;MTT方法检测肺癌细胞增殖水平,流式细胞术检测凋亡率的改变;蛋白［质］印迹法(Western blot)检测目的基因的蛋白表达变化。结果：与癌旁组织比较,HOTTIP在肺癌组织中的表达率明显增加,差异有统计学意义(P<0.05)。HOTTIP表达与患者淋巴结转移和TNM分期相关。转染HOTTIP siRNA至A549肺癌细胞后,结果显示,与未转染组和阴性质粒转染组相比,转染siRNA-HOTTIP 的A549细胞HOTTIP 表达明显降低;HOTTIP表达下调后细胞增殖速度减慢,凋亡率增加,凋亡相关蛋白Bax的表达增加,Bcl-2表达减少。结论：HOTTIP是一种新的NSCLC生物标志物,可作为潜在的治疗新靶点。     

lncRNA HOTTIP通过miR⁃637/KLK4轴促进肺癌SPC⁃A⁃1细胞的恶性 生物学行为

目的：探讨 lncRNA HOTTIP 对肺癌细胞增殖、凋亡及 EMT 的影响及其作用机制。方法：利用 qPCR 检测 lncRNAHOTTIP、miR-637和KLK4在肺癌SPC-A-1、正常肺上皮BEAS-2B细胞中的表达量;siRNA干扰SPC-A-1细胞中lncRNA HOTTIP的表达后,分别通过CCK-8、Transwell、流式细胞术和WB法检测SPC-A-1细胞增殖、侵袭、凋亡和EMT能力的变化。miRanda软件和双荧光素酶报告基因实验分析 lncRNA HOTTIP 和 miR-637 之间的靶向关系,RNA pull-down 实验检测 lncRNA HOTTIP 和miR-637的吸附作用,检测lncRNA HOTTIP通过miR-637对SPC-A-1细胞增殖、侵袭、凋亡和EMT的调控。利用TargetScan软件分析 miR-637 与 KLK4 的相关性,双荧光素酶报告基因实验检测 miR-637 与 KLK4 mRNA 之间的相互作用;检测 miR-637 通过KLK4 mRNA对SPC-A-1细胞增殖、侵袭、凋亡和EMT的调控。下调lncRNA HOTTIP和miR-637表达后,利用qPCR和WB检测KLK4 mRNA 和蛋白表达水平的变化。结果：与 BEAS-2B 细胞比,在 SPC-A-1 细胞中 lncRNA HOTTIP 呈高表达（P<0.01）,miR-637呈低表达（P<0.01）,KLK4呈高表达（P<0.01）。下调lncRNA HOTTIP后,SPC-A-1细胞增殖、侵袭与EMT能力显著减弱,细胞凋亡率显著上升（P<0.01）;lncRNA HOTTIP与miR-637具有靶向关系;下调miR-637表达后,SPC-A-1细胞增殖、侵袭与EMT能力显著上升,细胞凋亡率显著降低（P<0.01）。miR-637与 KLK4 3''UTR特异性结合。miR-637通过 KLK4显著促进了 SPC-A-1细胞增殖、侵袭与 EMT,细胞凋亡率显著上升（P<0.01）。下调 lncRNA HOTTIP 使 KLK4 表达显著降低,而下调 miR-637 可促进KLK4表达（P<0.05）。结论：上调lncRNA HOTTIP可通过miR-637/KLK4轴促进肺癌SPC-A-1细胞的增殖、侵袭与EMT而抑制癌细胞凋亡。     

Long non-coding RNA HOTTIP is up-regulated and associated with poor prognosis in patients with osteosarcoma

Long non-coding RNAs (lncRNAs) have been shown to play key roles in cancer development and progression. In this study, we focused on lncRNA HOTTIP and investigated its expression pattern, clinical significance, and biological function in osteosarcoma (OS). In the present study, lncRNA HOTTIP expression in OS tissues was examined and its correlation with clinicopathological features and patient prognosis was analyzed. In vitro assays were performed to understand the biological roles of lncRNA HOTTIP in OS progression. In the study, we found that HOTTIP expression was up-regulated in OS tissues, and correlated with advanced clinical stage and distant metastasis. OS patients with high HOTTIP expression level had poorer overall survival than those with low HOTTIP expression. Multivariable Cox proportional hazards regression analysis suggested that increased HOTTIP expression was an independent prognostic factor of overall survival in OS patients. Moreover, the results of in vitro assays showed that the suppression of HOTTIP in OS cells significantly reduced cell proliferation, migration and invasion ability. Our study demonstrated that lncRNA HOTTIP play critical roles in OS progression and could represent a novel prognostic marker and potential therapeutic target in OS patients.     

Epigenetic modulators from “The Big Blue”: A treasure to fight against cancer

Cancer remains a major public health problem in our society. The development of potent novel anti-cancer drugs selective for tumor cells is therefore still required. Deregulation of the epigenetic machinery including DNA methylation, histone modifications and non-coding RNAs is a hallmark of cancer, which provides potential new therapeutic targets. Natural products or their derivatives represent a major class of anti-cancer drugs in the arsenal available to the clinician. However, regarding epigenetically active anti-cancer agents for clinics, the oceans represent a largely untapped resource. This review focuses on marine natural compounds with epigenetic activities and their synthetic derivatives displaying anti-cancer properties including largazole, psammaplins, trichostatins and azumamides.     

不同级别子宫颈病变与lncRNA HOTTIP和H19单核苷酸多态性的关系研究

背景与目的:HOTTIP和H19都是lncRNA(long non-coding RNA,lncRNA)中的一员,在肿瘤的发生、发展和迁移过程中发挥重要作用.本研究探讨HOTTIP rs2067087、H19 rs2839698和H19 rs2107425单核苷酸多态性(single nucleotide polymo...