miR-618在非小细胞肺癌中的表达及临床意义

目的探讨微小RNA-618(miR-618)在非小细胞肺癌(NSCLC)中的表达水平及临床意义。方法收集2012年1月-2014年1月我院NSCLC患者手术标本85例,采用实时荧光定量PCR(qRT-PCR)检测miR-618在85例NSCLC组织和其配对的癌旁组织中的表达水平,Kaplan-Meier法绘制生存曲线,Log Rank检验分析高低表达水平组的生存率差异。分析miRNA-618的表达水平与NSCLC临床病理参数的关系;多因素Logistic回归分析miRNA-618表达水平的影响因素。结果miRNA-618在NSCLC组织中的表达为(0.78±0.48),显著低于癌旁组织中的表达水平(0.99±0.57)(P<0.05)。miR-618低表达的NSCLC患者的术后5年总生存率为15.22%(7/46),显著低于miR-618高表达的NSCLC患者的33.33%(13/39)。miR-618低表达组中位OS(<28个月),低于miR-618高表达组中位OS(>36个月)(P<0.05)。miR-618在NSCLC组织中的表达水平与患者的年龄、性别、组织类型无关(P>0.05),与肿瘤大小、分化程度、TNM分期和淋巴结转移有关(P<0.05)。Logistic回归显示:肿瘤大小、分化程度、TNM分期和淋巴结转移是miR-618表达的显著影响因素(P<0.05)。结论miRNA-618在NSCLC中表达下调,其表达与NSCLC发生发展、预后有关,miR-618可作为NSCLC诊断和预后预测新的靶点。     

血清CA125、miRNA-139-5p、miRNA-15a水平检测在卵巢癌患者中的临床意义

目的:探讨血清糖类抗原125(CA125)、微小RNA-139-5p(miRNA-139-5p)、微小RNA-15a(miRNA-15a)水平检测在卵巢癌患者中的临床意义。方法:选取74例卵巢癌患者作为卵巢癌组,选取同期74名该院健康体检者作为对照组,比较卵巢癌组与对照组、不同分化程度卵巢癌患者、不同病理分期卵巢癌患者血清CA125、miRNA-139-5p、miRNA-15a水平,分析血清CA125、miRNA-139-5p、miRNA-15a水平与卵巢癌分化程度和病理分期的相关性。结果:卵巢癌组CA125、miRNA-139-5p水平均高于对照组,miRNA-15a水平低于对照组,差异有统计学意义(P<0.05);卵巢癌低分化患者血清CA125、miRNA-139-5p水平均高于中分化和高分化患者,且中分化患者高于高分化患者;卵巢癌低分化患者miRNA-15a水平低于中分化和高分化患者,且中分化患者低于高分化患者,差异有统计学意义(P<0.05);卵巢癌患者血清CA125、miRNA-139-5p水平随着病理分期升高而升高,即Ⅰ期<Ⅱ期<Ⅲ期<Ⅳ期,卵巢癌患者miRNA-15a水平随着病理分期升高而降低,即Ⅰ期>Ⅱ期>Ⅲ期>Ⅳ期,差异有统计学意义(P<0.05);Pearson相关性分析结果显示,CA125、miRNA-139-5p水平与卵巢癌分化程度呈负相关,与病理分期呈正相关,miRNA-15a水平与卵巢癌分化程度呈正相关,与病理分期呈负相关(P<0.05)。结论:卵巢癌患者血清CA125、miRNA-139-5p水平升高,miRNA-15a水平降低,且与卵巢癌分化程度、病理分期存在相关性,血清CA125、miRNA-139-5p、miRNA-15a水平可为临床卵巢癌患者诊治提供参考依据。     

miRNA-30 b纳米粒转染对甲状腺未分化癌细胞系TA -K 细胞生物学特性的影响

目的：研究miRNA-30b纳米粒对甲状腺未分化癌细胞株TA-K细胞的生物学特性的影响。方法转染不同浓度的miRNA-30b纳米粒进入TA-K细胞中,通过流式法检测TA-K细胞凋亡情况,后通过生物信息学预测miR-NA-30b的生物学靶点,并通过RT-PCR及Western Blot方法验证。结果发现转染了miRNA-30b纳米粒后,TA-K细胞的凋亡显著增加,而生物信息学显示miRNA-30b可靶向作用于Survivin,后经RT-PCR及Western blot 法证实Sur-vivin确实是miRNA-30b的生物学靶点。结论 miRNA-30b可以通过作用Survivin促进TA-K细胞的凋亡,这一现象也为将来甲状腺未分化癌的临床治疗提供了新的思路。     

Liver biomarker and in vitro assessment confirm the hepatic origin of aminotransferase elevations lacking histopathological correlate in beagle dogs treated with GABAA receptor antagonist NP260

NP260 was designed as a first-in-class selective antagonist of α4-subtype GABA_A receptors that had promising efficacy in animal models of pain, epilepsy, psychosis, and anxiety. However, development of NP260 was complicated following a 28-day safety study in dogs in which pronounced elevations of serum aminotransferase levels were observed, although there was no accompanying histopathological indication of hepatocellular injury. To further investigate the liver effects of NP260, we assayed stored serum samples from the 28-day dog study for liver specific miRNA (miR-122) as well as enzymatic biomarkers glutamate dehydrogenase and sorbitol dehydrogenase, which indicate liver necrosis. Cytotoxicity assessments were conducted in hepatocytes derived from dog, rat, and human liver samples to address the species specificity of the liver response to NP260. All biomarkers, except ALT, returned toward baseline by Day 29 despite continued drug treatment, suggesting adaptation to the initial injury. In vitro analysis of the toxicity potential of NP260 to primary hepatocytes indicated a relative sensitivity of dog > human > rat, which may explain, in part, why the liver effects were not evident in the rodent safety studies. Taken together, the data indicate that a diagnostic biomarker approach, coupled with sensitive in vitro screening strategies, may facilitate interpretation of toxicity potential when an adaptive event masks the underlying toxicity.     

Circulating microRNA 122 in the methionine and choline‐deficient mouse model of non‐alcoholic steatohepatitis

Non‐alcoholic steatohepatitis (NASH) is a progressive form of non‐alcoholic fatty liver disease (NAFLD) and is a major cause of liver cirrhosis and hepatic failure. The methionine choline‐deficient diet (MCD) is a frequently used hepatotoxicity animal model of NASH that induces hepatic transaminase (ALT, AST) elevations and hepatobiliary histological changes similar to those observed in human NASH. Liver‐specific microRNA‐122 (miR‐122) has been shown as a key regulator of cholesterol and fatty acid metabolism in adult liver, and has recently been proposed as a sensitive and specific circulating biomarker of hepatic injury. The purpose of this study was to assess miR‐122 serum levels in mice receiving an MCD diet for 0, 3, 7, 14, 28 and 56 days and compare the performance vs. routine clinical chemistry when benchmarked against the histopathological liver findings. MiR‐122 levels were quantified in serum using RT‐qPCR. Both miR‐122 and ALT/AST levels were significantly elevated in serum at all timepoints. MiR‐122 levels increased on average by 40‐fold after 3 days of initiating the MCD diet, whereas ALT and AST changes were 4.8‐ and 3.3‐fold, respectively. In general, miR‐122 levels remained elevated across all time points, whereas the ALT/AST increases were less robust but correlated with the progressive severity of NASH as assessed by histopathology. In conclusion, serum levels of miR‐122 can potentially be used as a sensitive biomarker for the early detection of hepatotoxicity and can aid in monitoring the extent of NAFLD‐associated liver injury in mouse efficacy models. Copyright © 2013 John Wiley & Sons, Ltd.     

非瓣膜病心房颤动患者循环miRNA-21水平及应用价值初探

目的：探索非瓣膜病心房颤动患者循环miRNA-21的水平及可能的应用价值。 方法：前瞻性的筛选连续收治的非瓣膜病心房颤动患者,收集患者的人口学特征,病史等资料。将CHA2DS2-VASc评分≥2分（男）或≥3分（女）定义为高卒中风险。采集患者空腹外周静脉血,ELISA法检测NT-proBNP;qRT-PCR法检测循环miRNA-21水平。评估miRNA-21水平与CHA2DS2-VASc评分和NT-proBNP之间的相关性,ROC曲线验证miRNA-21对高卒中风险的预测价值。 结果：共入选患者89例,高卒中风险患者41例（46.1%）,中位CHA2DS2-VASc评分4（2.5-5.5）分。,高卒中风险组与低卒中风险组血常规、肝肾功能、NT-proBNP等无显著差异;高卒中风险组循环miRNA-21表达量显著升高。Pearson相关分析显示循环miRNA-21表达量与CHA2DS2-VASc评分(r=0.461,p=0.002)和NT-proBNP（r=0.386, p=0.013）均显著相关。ROC分析提示miRNA-21表达量对高卒中风险具有较高预测价值（AUC=0.843, p＜0.001）。 结论：循环miRNA-21可能用于预测非瓣膜病心房颤动患者高卒中风险。     

Phospholipase D1 inhibition sensitizes glioblastoma to temozolomide and suppresses its tumorigenicity

Resistance of glioblastoma to the chemotherapeutic compound temozolomide is associated with the presence of glioblastoma stem cells in glioblastoma and is a key obstacle for the poor prognosis of glioblastoma. Here, we show that phospholipase D1 is elevated in CD44^High glioblastoma stem cells and in glioblastoma, especially recurring glioblastoma. Phospholipase D1 elevation positively correlated with the level of CD44 and poor prognosis in glioblastoma patients. Temozolomide significantly upregulated the expression of phospholipase D1 in the low and moderate CD44 populations of glioblastoma stem cells, but not in the CD44^High population in which phospholipase D1 is highly expressed. Phospholipase D1 conferred resistance to temozolomide in CD44^High glioblastoma stem cells and increased their self-renewal capacity and maintenance. Phospholipase D1 expression significantly correlated with levels of temozolomide resistance factors, which were suppressed by microRNA-320a and -4496 induced by phospholipase D1 inhibition. Genetic and pharmacological targeting of phospholipase D1 attenuated glioblastoma stem cell-derived intracranial tumors of glioblastoma using the microRNAs, and improved survival. Treatment solely with temozolomide produced no benefits on the glioblastoma, whereas in combination, phospholipase D1 inhibition sensitized glioblastoma stem cells to temozolomide and reduced glioblastoma tumorigenesis. Together, these findings indicate that phospholipase D1 inhibition might overcome resistance to temozolomide and represents a potential treatment strategy for glioblastoma. © 2020 The Authors. The Journal of Pathology published by John Wiley & Sons, Ltd. on behalf of The Pathological Society of Great Britain and Ireland.