Analysis of anti‐apoptotic PVT1 oncogene and apoptosis‐related proteins (p53, Bcl2, PD‐1, and PD‐L1) expression in thyroid carcinoma

BackgroundAn aberrant expression of long non‐coding RNA PVT1 has been associated with apoptosis in various cancer types. We aimed to explore the PVT1 and four apoptosis‐related proteins (p53, Bcl2, and PD‐1/PD‐L1) signature in thyroid cancer (TC).MethodsThe PVT1 expression level was measured in 64 FFPE TC paired samples by real‐time quantitative PCR. Overall and stratified analyses by different clinicopathological features were done. The apoptotic proteins were evaluated by immunohistochemistry staining.ResultsOverall analysis showed significant PVT1upregulation in TC tissues (p < 0.001). Similarly, subgroup analysis by BRAF ^V600E mutation showed consistent results. Lower expression of p53 was associated with mortality (p = 0.001). Bcl2 overexpression was associated with greater tumor size (p = 0.005). At the same time, HCV‐positive cases were associated with repressed Bcl2 expression levels (54.3% in HCV‐negative vs. 6.9% in HCV‐positive cases, p = 0.011). PD‐1 expression was associated with lymph node metastasis (p = 0.004). Enhanced PD‐L1 expression in the tumor was associated with a higher tumor stage, lymphovascular invasion, and mortality risk. Kaplan–Meier curves for overall survival showed that low p53 and high PD‐L1 expressions were associated with lower survival time. The p53‐positive staining is associated with a 90% decreased mortality risk (HR = 0.10, 95%CI = 0.02–0.47, p = 0.001), while patients with high PD‐L1 were five times more likely to die (HR = 4.74, 95%CI = 1.2–18.7, p = 0.027).ConclusionOur results confirm the upregulation of PVT1 in TC. The apoptosis‐related proteins (p53, Bcl2, and PD‐1/PD‐L1) showed different prognostic utility in TC patients; in particular, low p53 and high PD‐L1 expressions associated with low survival times. Further large‐scale and mechanistic studies are warranted.     

Quercetin regulates inflammation,oxidative stress,apoptosis, and mitochondrial structure and function in H9C2 cells by promoting PVT1 expression

ObjectiveTo investigate the effect and potential mechanism of quercetin on inflammation, oxidative stress, apoptosis, and mitochondrial structure and function in H9C2 cells.Materials and methodsH9C2 cells were obtained from the Shanghai Institutes for Biological Sciences, Chinese Academy of Science, and randomly divided into six groups: control, model, PVT1 overexpression (OV), quercetin, OV + quercetin, and NAC groups. The CCK-8 assay was performed to examine cell proliferation. Flow cytometry was used to examine cell apoptosis, cell membrane potential, and ROS levels. The expression of endothelial nitric oxide synthase (eNOS), malondialdehyde (MDA), and superoxide dismutase (SOD) was measured by ELISA and a Biochemical kit. Western blotting was used to determine the levels of p-DRP1 (s637), MFN2, NF-kB, p-NF-kB, IkB, and p-IkB. IL-6, IL-10, TNF-α, and IL-1β mRNA expression was examined by RT-PCR. Electron microscopy was used to observe the structure of mitochondria in H9C2 cells.ResultsMDA, p-NF-κB, p-IKB, IL-6, IL-1β, and TNF-α expression levels, and the cell apoptosis rate were significantly higher in the model group than in the control group (P < 0.05). In contrast, the cell proliferation rate and IL-10, SOD, eNOS, and ATP levels were significantly lower in the model group (P < 0.05). Moreover, MDA expression was significantly lower in the OV, quercetin, quercetin + OV, and NAC groups than in the model group (P < 0.05), while SOD, eNOS, and ATP levels were higher. The electron microscopy results showed that PVT1 overexpression or quercetin treatment could inhibit inflammation-induced mitochondrial fission and promote mitochondrial fusion.ConclusionQuercetin promotes the proliferation of H9C2 cells, while inhibiting inflammation, oxidative stress, and cell apoptosis, and alleviating the structural and functional dysfunction of mitochondria. These effects are achieved by promoting PVT1 expression.     

长链非编码RNA PVT1通过激活Wnt信号通路对肝癌干细胞自我更新能力的调控

摘 要 目的：探讨长链非编码RNA PVT1通过激活Wnt信号通路对肝癌干细胞自我更新能力的调控作用。 方法: 设HEPG2肝癌干细胞组、LNCRNA PVT1 mimics组、LNCRNA PVT1 inhibitor组,培养结束后,检测各组细胞活力、单克隆形成数目、成球率、凋亡率水平、G1期、PVT1 mRNA、WNT、SWI、SNF蛋白表达水平。 结果: LNCRNA PVT1 mimics组OD值、存活率水平、克隆形成数目、成球率、G1期、PVT1 mRNA、WNT、SWI、SNF蛋白表达水平高于HEPG2肝癌干细胞组（P<0.05）,细胞凋亡率低于HEPG2肝癌干细胞组（P<0.05）;LNCRNA PVT1 inhibitor组OD值、存活率、克隆形成数目、成球率、G1期、PVT1 mRNA、WNT、SWI、SNF蛋白表达水平低于HEPG2肝癌干细胞组、LNCRNA PVT1 mimics 组（P<0.05）,细胞凋亡率高于HEPG2肝癌干细胞组、LNCRNA PVT1 mimics 组(P<0.05)。LNCRNA PVT1 mRNA 与克隆形成数目、成球率、G1期、WNT、SWI、SNF正相关关系明显,与凋亡率负相关关系明显（P<0.05）。 结论: LNCRNA PVT1通过激活SWI/SNF复合物进而激活Wnt信号传导来促进肝癌干细胞的自我更新和肿瘤增殖。     

Increased expression of the lncRNA PVT1 promotes tumorigenesis in non-small cell lung cancer

Introduction: Non-small cell lung cancer (NSCLC) is the major cause of cancer death worldwide. Increasing evidence shows that long non coding RNAs (lncRNAs) are widely involved in the development and progression of NSCLC. lncRNA PVT1 in several cancers has been studied, its role in lung cancer remains unknown. Our studies were designed to investigate the expression, biological role and clinical significance of PVT1 in lung cancer. Methods: lncRNA PVT1 expression in 82 NSCLC tissues and 3 lung cancer cell lines was measured by quantitative Real-time PCR (qRT-PCR). Its association with overall survival of patients was analyzed by statistical analysis. RNA interference (RNAi) approaches were used to investigate the biological functions of PVT1. The effect of PVT1 on proliferation was evaluated by MTT, cell migration and invasion ability was evaluated by cell migration and invasion assays. Results: lncRNA PVT1 expression was significantly upregulated in NSCLC tissues and lung cancer cells when compared with corresponding adjacent normal tissues and normal bronchial epithelial cells. Increased PVT1 expression was significantly correlated with histological grade and lymph node metastasis. In addition, NSCLC patients with PVT1 higher expression have shown significantly poorer overall survival than those with lower PVT1 expression. And PVT1 expression was an independent prognostic marker of overall survival in a multivariate analysis. In vitro assays our results indicated that knockdown of PVT1 inhibited cell proliferation, migration, and invasion. Conclusions: Our data indicated that lncRNA PVT1 is significantly upregulated in NSCLC tissues and may represent a new biomarker and a potential therapeutic target for NSCLC intervention.     

Evaluating the potency of blood long noncoding RNA PVT1 as candidate biomarker reflecting inflammation,multiple organ dysfunction,and mortality risk in sepsis patients

ObjectiveLong noncoding RNA plasmacytoma variant translocation 1 (lnc‐PVT1) promotes septic inflammation and organ injuries via multiple ways, while its clinical engagement in sepsis management is indistinct. This study aimed to investigate its relationship with inflammation, multiple organ dysfunction, and mortality risk in sepsis patients.MethodsSepsis patients and age‐/gender‐matched healthy controls were enrolled; their lnc‐PVT1 expression in plasma were detected by RT‐qPCR. For sepsis patients only, the inflammatory cytokine levels (tumor necrosis factor (TNF)‐α, interleukin (IL)‐1β, and IL‐17A) in plasma were detected by ELISA. According to the survival data during 28‐day follow‐up, sepsis patients were divided into sepsis survivors and sepsis deaths.ResultsLnc‐PVT1 expression was increased in sepsis patients (N = 157) compared with healthy controls (N = 80) (p < 0.001). In sepsis patients, lnc‐PVT1 was linked with higher acute physiology and chronic health evaluation II (APACHEII) score (p = 0.001), total sequential organ failure assessment (SOFA) score, and its most subitems (SOFA‐respiratory system, SOFA‐coagulation, SOFA‐liver, SOFA‐cardiovascular system, and SOFA‐renal system scores) (all p < 0.01), but not SOFA‐nervous system score (p = 0.091); it did not relate to primary infection sites either (p = 0.204). Furthermore, lnc‐PVT1 correlated with increased C‐reactive protein, TNF‐α, IL‐1β, and IL‐17 in sepsis patients (all p < 0.01). Additionally, lnc‐PVT1 expression was higher in sepsis deaths than that in sepsis survivors (p < 0.001), following receiver‐operating characteristic curve disclosed that lnc‐PVT1 predicted 28‐day septic mortality risk (area under the curve: 0.789, 95% confidence interval: 0.702–0.875).ConclusionCirculating lnc‐PVT1 exhibits the potential as a biomarker in sepsis patients to inform inflammation, multiple organ dysfunction, and mortality risk.     

Oral Anticoagulation in Patients With Liver Disease

Patients with liver disease are at increased risks of both thrombotic and bleeding complications. Many have atrial fibrillation (AF) or venous thromboembolism (VTE) necessitating oral anticoagulant agents (OACs). Recent evidence has contradicted the assumption that patients with liver disease are “auto-anticoagulated” and thus protected from thrombotic events. Warfarin and non–vitamin K–antagonist OACs have been shown to reduce thrombotic events safely in patients with either AF or VTE. However, patients with liver disease have largely been excluded from trials of OACs. Because all currently approved OACs undergo metabolism in the liver, hepatic dysfunction may cause increased bleeding. Thus, the optimal anticoagulation strategy for patients with AF or VTE who have liver disease remains unclear. This review discusses pharmacokinetic and clinical studies evaluating the efficacy and safety of OACs in patients with liver disease and provides a practical, clinically oriented approach to the management of OAC therapy in this population.     

PVT1 induces NSCLC cell migration and invasion by regulating IL-6 via sponging miR-760

Non-small-cell lung carcinoma (NSCLC) accounts for approximately 80% of lung cancers with a high metastatic potential. Elucidating the mechanism of NSCLC metastasis will provide new promising targets for NSCLC therapy and benefit its prognosis. Plasmacytoma variant translocation 1 (PVT1) has been proven to be overexpressed in NSCLC. Although the oncogenic role of PVT1 in NSCLC has been reported, its mechanism remains unclear. Here, we verified that the knockdown of PVT1 inhibited NSCLC cell migration and invasion, and that its inhibitory role on A549 cells and H1299 cells was antagonized by interleukin-6 (IL-6) treatment. The results revealed that PVT1 regulates IL-6 by sponging miR-760 and identified the binding site of miR-760 in the 3′-UTR of IL-6. In conclusion, a new mechanism was revealed, wherein PVT1 regulates NSCLC cell migration and invasion via miR-760/IL-6, suggesting PVT1/miR-760/IL-6 as promising prognostic biomarkers and therapeutic targets for NSCLC metastasis.     

Neurobehavioral Dynamics Following Chronic Sleep Restriction: Dose-Response Effects of One Night for Recovery

Objective:

Establish the dose-response relationship between increasing sleep durations in a single night and recovery of neurobehavioral functions following chronic sleep restriction.

Design:

Intent-to-treat design in which subjects were randomized to 1 of 6 recovery sleep doses (0, 2, 4, 6, 8, or 10 h TIB) for 1 night following 5 nights of sleep restriction to 4 h TIB.

Setting:

Twelve consecutive days in a controlled laboratory environment.

Participants:

N = 159 healthy adults (aged 22-45 y), median = 29 y).

Interventions:

Following a week of home monitoring with actigraphy and 2 baseline nights of 10 h TIB, subjects were randomized to either sleep restriction to 4 h TIB per night for 5 nights followed by randomization to 1 of 6 nocturnal acute recovery sleep conditions (N = 142), or to a control condition involving 10 h TIB on all nights (N = 17).

Measurements and Results:

Primary neurobehavioral outcomes included lapses on the Psychomotor Vigilance Test (PVT), subjective sleepiness from the Karolinska Sleepiness Scale (KSS), and physiological sleepiness from a modified Maintenance of Wakefulness Test (MWT). Secondary outcomes included psychomotor and cognitive speed as measured by PVT fastest RTs and number correct on the Digit Symbol Substitution Task (DSST), respectively, and subjective fatigue from the Profile of Mood States (POMS). The dynamics of neurobehavioral outcomes following acute recovery sleep were statistically modeled across the 0 h-10 h recovery sleep doses. While TST, stage 2, REM sleep and NREM slow wave energy (SWE) increased linearly across recovery sleep doses, best-fitting neurobehavioral recovery functions were exponential across recovery sleep doses for PVT and KSS outcomes, and linear for the MWT. Analyses based on return to baseline and on estimated intersection with control condition means revealed recovery was incomplete at the 10 h TIB (8.96 h TST) for PVT performance, KSS sleepiness, and POMS fatigue. Both TST and SWE were elevated above baseline at the maximum recovery dose of 10 h TIB.

Conclusions:

Neurobehavioral deficits induced by 5 nights of sleep restricted to 4 h improved monotonically as acute recovery sleep dose increased, but some deficits remained after 10 h TIB for recovery. Complete recovery from such sleep restriction may require a longer sleep period during 1 night, and/or multiple nights of recovery sleep. It appears that acute recovery from chronic sleep restriction occurs as a result of elevated sleep pressure evident in both increased SWE and TST.

Citation:

Banks S; Van Dongen HPA; Maislin G; Dinges DF. Neurobehavioral dynamics following chronic sleep restriction: dose-response effects of one night for recovery. SLEEP 2010;33(8):1013–1026.     

长链非编码RNA PVT1调控miR-551通过Wnt信号通路对卵巢癌迁移和侵袭的影响

目的:探讨长链非编码RNA PVT1在卵巢癌组织中的表达情况及其在卵巢癌细胞迁移和侵袭过程中的作用及机制。方法:q PCR检测卵巢癌和正常卵巢组织及不同卵巢癌细胞中PVT1的表达情况;Transwell侵袭实验和细胞划痕实验分别检测沉默PVT1后卵巢癌细胞侵袭和迁移能力的变化;双萤光素酶报告基因检测PVT1与微小RNA(miR)-551的相互作用;Transwell侵袭实验和细胞划痕实验分别检测沉默PVT1后miR-551-inhibitor对卵巢癌细胞侵袭和迁移能力的影响;Western blot法检测沉默PVT1后Wnt信号通路相关蛋白的表达情况。裸鼠皮下成瘤实验检测沉默PVT1对卵巢癌成瘤重量及体积的影响。结果:与正常卵巢组织相比,卵巢瘤组织中PVT1表达明显增高(P0.05);卵巢癌细胞株ES-2中PVT1表达水平最高(P0.05);沉默PVT1可以抑制卵巢癌细胞侵袭和迁移能力;PVT1能与miR-551的位点特异性结合;沉默PVT1后,miR-551-inhibitor可以促进卵巢癌细胞侵袭和迁移能力;沉默PVT1后Wnt信号通路蛋白的表达相应下调;与阴性对照组相比,PVT1-siRNA组荷瘤小鼠肿瘤体积和重量都明显减小(P0.05)。结论:PVT1在卵巢癌发生发展过程中起重要作用,它可以靶向调节miR-551,通过Wnt信号通路调控卵巢癌细胞的侵袭和迁移能力。