ATP6L promotes metastasis of colorectal cancer by inducing epithelial‐mesenchymal transition

ATP6L, the C subunit of the V‐ATPase V0 domain, is involved in regulating the acidic tumor micro‐environment and may promote tumor progression. However, the expression and functional role of ATP6L in tumors have not yet been well explored. In this study, we found that ATP6L protein overexpression was related to colorectal cancer histological differentiation (P < 0.001), presence of metastasis (P < 0.001) and recurrence (P = 0.02). ATP6L expression in the liver metastatic foci was higher than in the primary foci (P = 0.04). ATP6L expression was notably concomitant with epithelial‐mesenchymal transition (EMT) immunohistochemical features, such as reduced expression of the epithelial marker E‐cadherin (P = 0.021) and increased expression of the mesenchymal marker vimentin (P = 0.004). Results of in vitro and in vivo experiments showed that ATP6L expression could alter cell morphology, regulate EMT‐associated protein expression, and enhance migration and invasion. The effect of ATP6L on metastasis was further demonstrated in a tail vein injection mice model. In addition, the mouse xenograft model showed that ATP6L‐overexpressing HCT116 cells grew into larger tumor masses, showed less necrosis and formed more micro‐vessels than the control cells. Taken together, our results suggest that ATP6L promotes metastasis of colorectal cancer by inducing EMT and angiogenesis, and is a potential target for tumor therapy.     

Kaposiform lymphangiomatosis treated with multimodal therapy improves coagulopathy and reduces blood angiopoietin‐2 levels

Kaposiform lymphangiomatosis (KLA) is a rare, life‐threatening congenital lymphatic malformation. Diagnosis is often delayed due to complex indistinct symptoms. Blood angiopoietin‐2 (ANG2) levels are elevated in KLA and may be useful as a biomarker to monitor disease status. We report a 7‐year‐old male child with easy bruising, inguinal swelling, and consumptive coagulopathy, diagnosed with KLA. A multimodal treatment regimen of prednisone, sirolimus, vincristine, and adjunctive zoledronate was used. Plasma ANG2 levels were highly elevated at diagnosis but decreased during treatment. The patient showed significant clinical improvement over a 38‐month period and normalization of ANG2 levels correlated with resolution of the coagulopathy.     

Vasogenic edema versus neuroplasticity as neural correlates of hippocampal volume increase following electroconvulsive therapy

BackgroundVolume increases of the hippocampus after electroconvulsive therapy (ECT) are a robust finding, pointing into the direction of neurogenesis. However, such volumetric increases could also be explained by edema and/or neuroplastic changes (such as angiogenesis).ObjectivesIf edema explains the volume increase of the hippocampus we hypothesize it would lead to increased mean diffusivity (MD). If neuroplastic would explain the volume increase, it would lead to decreased MD. To investigate angiogenesis as explanation we studied the perfusion fraction f and the pseudodiffusion component D1 obtained from intravoxel incoherent motion (IVIM) data, and relative perfusion changes obtained from arterial spin labelling (ASL) data.MethodsUsing ultra-high field (7 tesla) MRI we acquired IVIM and ASL data. We compared MD, f, D1 and ASL values for both hippocampi in 21 patients (before and after 10 ECT sessions) and 8 healthy controls (without ECT) in a linear mixed model adjusting for age and gender.ResultsWe found a significant decrease in MD (which was absent in the healthy controls) in the left and right hippocampus (t = -3.98, p < 0.001). In addition, a decrease in f (t = -4.61, p < 0.001, but not in controls) and no differences in D1 or ASL perfusion values (both p > 0.05) were found.ConclusionsThe decrease in MD in perfusion fraction f suggest that formation of edema nor angiogenesis are responsible for the ECT-induced volume increases in the hippocampus. Also, it supports the hypothesis that hippocampal volume increases might be due to neuroplastic changes.     

RPV‐modified epirubicin and dioscin co‐delivery liposomes suppress non‐small cell lung cancer growth by limiting nutrition supply

Chemotherapy for non‐small cell lung cancer (NSCLC) is far from satisfactory, mainly due to poor targeting of antitumor drugs and self‐adaptations of the tumors. Angiogenesis, vasculogenic mimicry (VM) channels, migration, and invasion are the main ways for tumors to obtain nutrition. Herein, RPV‐modified epirubicin and dioscin co‐delivery liposomes were successfully prepared. These liposomes showed ideal physicochemical properties, enhanced tumor targeting and accumulation in tumor sites, and inhibited VM channel formation, tumor angiogenesis, migration and invasion. The liposomes also downregulated VM‐related and angiogenesis‐related proteins in vitro. Furthermore, when tested in vivo, the targeted co‐delivery liposomes increased selective accumulation of drugs in tumor sites and showed extended stability in blood circulation. In conclusion, RPV‐modified epirubicin and dioscin co‐delivery liposomes showed strong antitumor efficacy in vivo and could thus be considered a promising strategy for NSCLC treatment.     

Protease‐activated receptor‐2 accelerates intestinal tumor formation through activation of nuclear factor‐κB signaling and tumor angiogenesis in ApcMin/+ mice

Hepatocyte growth factor activator inhibitor‐1 (HAI‐1), encoded by the SPINT1 gene, is a membrane‐bound protease inhibitor expressed on the surface of epithelial cells. Hepatocyte growth factor activator inhibitor‐1 regulates type II transmembrane serine proteases that activate protease‐activated receptor‐2 (PAR‐2). We previously reported that deletion of Spint1 in Apc^Min/+ mice resulted in accelerated formation of intestinal tumors, possibly through enhanced nuclear factor‐κB signaling. In this study, we examined the role of PAR‐2 in accelerating tumor formation in the Apc^Min/+ model in the presence or absence of Spint1. We observed that knockout of the F2rl1 gene, encoding PAR‐2, not only eliminated the enhanced formation of intestinal tumors caused by Spint1 deletion, but also reduced tumor formation in the presence of Spint1. Exacerbation of anemia and weight loss associated with HAI‐1 deficiency was also normalized by compound deficiency of PAR‐2. Mechanistically, signaling triggered by deregulated protease activities increased nuclear translocation of RelA/p65, vascular endothelial growth factor expression, and vascular density in Apc^Min/+‐induced intestinal tumors. These results suggest that serine proteases promote intestinal carcinogenesis through activation of PAR‐2, and that HAI‐1 plays a critical tumor suppressor role as an inhibitor of matriptase, kallikreins, and other PAR‐2 activating proteases.     

miR-200、 miR -155及血管新生因子与复发性流产的相关性分析

目的 研究miR-200、miR-155及血管新生因子与原因不明复发性流产(unexplained recurrent spontaneousabortion,URSA)的相关性分析。 方法 选择2015年3月—2018年1月在青岛市妇女儿童医院妇产科就诊的URSA患者作为URSA组、要求终止妊娠的正常早孕患者作为对照组,检测绒毛组织中微小RNA(microRNA, miR)miR-200、miR-155、血管内皮生长因子(vascular endothelial growth factor,VEGF)、可溶性FMS样酪氨酸激酶1(soluble FMS like tyrosine kinase-1,sFlt-1)的表达量及血清中VEGF、sFlt-1的含量,对miR-200、miR-155靶向结合VEGF、sFlt-1进行生物信息学分析。 结果 URSA组的绒毛组织中miR-200(1.78±0.32 vs. 0.91±0.15)、sFlt-1(1.87±0.35 vs. 1.06±0.21)的相对表达量及血清中sFlt-1的含量[(12.39±2.31)ng/ml vs. (6.51±0.95)ng/ml]均高于对照组,差异有统计学意义(均P<0.05)。绒毛组织中miR-155相对表达量(0.60±0.10 vs. 0.93±0.16)、VEGF mRNA相对表达量(0.59±0.09 vs. 1.02±0.16)及蛋白表达量(0.62±0.07 vs. 1.04±0.18)、血清中VEGF的含量[(601.25±94.39)ng/ml vs. (935.12±132.47)ng/ml]低于对照组,差异有统计学意义(均P<0.05);URSA组患者绒毛组织中miR-200的表达量与血清中VEGF的含量、绒毛组织中VEGF的表达量均呈负相关,绒毛组织中miR-155的表达量与血清中sFlt-1的含量和绒毛组织中sFlt-1的表达量均呈负相关;miR-200、miR-155分别靶向结合VEGF、sFlt-1基因的3’UTR。 结论 miR-200表达增多、miR-155表达减少与URSA发生有关,miR-200靶向VEGF、miR-155靶向sFlt-1是介导该过程的可能机制。     

通窍活血汤含药脑脊液对OGD/R损伤大鼠BMECs的保护作用

目的:探讨通窍活血汤含药脑脊液对氧糖剥夺/复糖复氧(OGD/R)损伤大鼠脑微血管内皮细胞(BMECs)的保护作用及潜在机制。方法:通过酶消化法提取原代BMECs,并将细胞随机分为6组,分别为正常组,OGD/R组,通窍活血汤(TQHXT)组(20%),尼莫地平(NMDP)组(10μmol·L~(-1)),卡博替尼(BMS)组(1μmol·L~(-1))和合用药组。除正常组外,其余各组细胞在氧糖剥夺2 h后迅速复糖复氧24 h进行OGD/R造模并分组给药。采用细胞免疫荧光染色法鉴定BMECs,观察OGD/R损伤大鼠BMECs的形态学和超微结构改变并检测细胞跨膜电阻(TEER)值变化。用试剂盒检测细胞内一氧化氮(NO)水平、乳酸脱氢酶(LDH)活性、活性氧(ROS)荧光强度和组织型纤溶酶原激活因子(tPA)含量。采用流式细胞术检测细胞内钙离子浓度及细胞凋亡,观察血管新生标记因子CD34的表达,用蛋白免疫印迹法(Western blot)检测细胞中紧密连接蛋白(ZO-1),血管内皮生长因子(VEGF),黏着斑激酶(FAK)和桩蛋白(Paxillin)蛋白的表达情况。结果:与正常组比较,OGD/R组细胞皱缩、变圆,细胞TEER值和细胞中ZO-1蛋白表达显著降低,细胞中NO,LDH及ROS水平显著升高,tPA含量显著降低,细胞中钙离子浓度和细胞凋亡显著增加,细胞中CD34有所表达,VEGF,FAK和Paxillin蛋白表达显著升高(P0. 01);与OGD/R组比较,TQHXT组细胞损伤明显改善,细胞TEER值和细胞中ZO-1蛋白表达显著升高,细胞中NO,LDH及ROS含量显著降低,tPA含量显著升高,细胞中钙离子浓度和细胞凋亡显著减少,细胞中CD34表达增加,VEGF,FAK和Paxillin蛋白表达显著升高(P0. 05,P0. 01)。结论:通窍活血汤含药脑脊液对OGD/R损伤大鼠BMECs具有保护作用,该保护作用可能是通过VEGF/VEGF受体2(R2)/FAK/Paxillin信号通路促进血管生成来发挥作用。     

中药复方肠复康抗结直肠癌血管生成作用靶基因网络药理学研究

目的运用网络药理学方法,研究中药复方肠复康(CFK)对结直肠癌(colorectal cancer,CRC)血管生成作用靶基因,为揭示其作用机制奠定理论基础。方法采用TTD、DrugBank数据库、OMIM数据库、GAD和PharmGKB等5个数据库分别检索CRC基因;采用TSMSP数据库及基于VBA工具的有效成分筛选方式检索CFK所含的5味中药,利用ADME参数进行有效成分筛选,通过TCMSP数据库检索各个有效成分的靶基因,利用cytoscape 3.2.1软件及其插件ClueGO、Bisogenet、CytoNCA对靶基因进行分析并构建CFK成分靶点网络图,并结合KEGG数据库进一步说明CFK与CRC血管生成靶基因的关系。结果 CRC靶基因339个,CFK靶基因182个;通过cytoscape构建并结合网络拓扑分析,ADRA1A、ADRA1B、ADRA1D、ADRB1、CHRM1、CHRM2、CHRM3、CHRM4、INSR、PIK3CG、RXRA、BAX、BCL2、CASP8、ICAM1、NFKBIA、CASP9、KDR、MAP2、PRKCA、PTGS2等靶点与血管生成相关;CFK治疗CRC过程中BRCA1、CDK2、CDKN1A、ITGA4、MDM2、YWHAG、CREBBP、CUL2、EP300、VHL、FLNA、SHC1、TRAF6、XPO1、EGFR、GRB2、IKBKG、NTRK1、TP53、 YWHAB、YWHAE与血管生成相关;主要通过调控PI3K-Akt、MAPK、HIF-1及VEGF信号通路抑制CRC患者新生血管生成。结论 CFK可能会通过VBRCA1、CDK2、CDKN1A、ITGA4、MDM2、YWHAG、CREBBP、CUL2、EP300、VHL、FLNA、SHC1、TRAF6、XPO1、EGFR、GRB2、IKBKG、NTRK1、TP53、 YWHAB、YWHAE等靶点调控PI3K-Akt、MAPK、HIF-1及VEGF等信号通路,进而发挥抑制CRC患者新生血管生成作用,具体机制有待进一步研究。