Giganteaside D induces ROS-mediated apoptosis in human hepatocellular carcinoma cells through the MAPK pathway

Purpose

Every year, almost one million individuals are diagnosed with hepatocellular carcinoma (HCC) worldwide and more than 690,000 patients die of it. At present, most therapeutic anti-HCC agents are not effective, which is due to the appearance of chemo-resistance and/or toxic side effects. Therefore, it is imperative to find novel more effective anti-HCC agents. Here, we evaluated the effect of giganteaside D (GD), an oleanolic acid saponin from P. scabiosaefolia, on the growth and apoptosis of HCC cells.

Methods and results

Using MTT and clonogenic assays, we found that GD exhibited a significant growth inhibitory effect on the HCC-derived cell lines HepG2 and Bel-7402. In addition, we found that GD induced mitochondria-mediated apoptosis in these HCC-derived cells, as indicated by a decreased mitochondrial potential, activation of Caspase-9 and Caspase-3, cleavage of PARP and release of Cytochrome C from the mitochondria. Besides, we found that GD stimulated the generation of reactive oxygen species (ROS) and that blockage of ROS attenuated the GD-induced mitochondria-mediated apoptosis. Additionally, we found that GD treatment led to a decrease in phosphorylated Erk (p-Erk) and triggered the generation of p-JNK, both components of the mitogen-activated protein kinase (MAPK) signaling pathway. Inhibition of Erk or JNK by specific inhibitors or siRNAs augmented or attenuated the cytotoxic and apoptotic effects of GD.

Conclusions

From our results we conclude that GD can induce ROS-mediated apoptosis in HCC-derived cells through the MAPK pathway. This observation may open up avenues to explore the future use of GD as a HCC chemotherapeutic agent.

     

Detection and clinical significance of thrombomodulin in both plasma and tissue extracts of cancer patients

Thrombomodulin?is the endothelial cell surface glycoprotein that converts thrombin from a procoagulant protein into the activator for protein C. Thrombin plays an important role in the adhesion of cancer cells to endothelial cells through fibrin formation and platelet activation, and thrombin stimulates tumor cell bindings to endothelial cells[1]. In this article, the changes of TM were studied in both plasma and tissue extracts of cancer patients for evaluating its clinical significance. P…     

贲门癌术前大剂量放疗与单纯手术的比较(附60例报告)

我院胸外科在1987年3月至1989年3月随机性对Ⅲ期贲门癌30例行术前10MeVX线的直线加速器、10cm×10cm的前后两野快速、大剂量（25Gy／5次／5天）对穿照射，共和30例单纯手术的结果进行比较。组织学的有效率64％。放疗组肿瘤切除率为83．3％（25／30），单纯手术组为80％（24／30）。5年生存率放疗组30％（9／30），单纯手术组23.3％（7／30）。两组比较无显著性差异（p＞0．05）。术后并发症前者未增加。     

重组人肿瘤坏死因子β(HuTNFβ)的纯化及抗肿瘤效应的初步研究

本文利用能表达HuTNFβ的工程菌株(P20KLT)按常规培养及扩增．即采用LB培养液(内含Amp100．μg／ml Tet 5μg／ml)；1：40扩增．当A550达0．1时，加入IAA(20μg／ml)，当A550为1．0时，中止培养，离心(5000rpm，4℃)收集菌体。在菌体中按1：5加入缓冲液，冰浴中超声破碎，13，000rpm高速离心20分钟，     

Targeting an RNA-Binding Protein Network in Acute Myeloid Leukemia

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TLR3和TLR4基因多态性与EBV相关胃癌易感性的关系

目的 探讨TLR3c.1377和TLR4Asp299Gly基因多态性与EBV感染在EBV相关胃癌（EBVassociated gastric carcinoma, EBVaGC）发生中的相关性。方法 选用41例EBVaGC组织、62例EBV阴性胃癌（EBV-negative gastric carcinoma, EBVnGC）组织以及64例健康人群血标本作为研究对象,采用PCR结合限制性长度多态性分析（reaction-restriction fragment length polymorphism, RFLP)技术检测TLR3c.1377 和TLR4 Asp299Gly基因多态性,并对实验结果进行统计分析。结果 （1）胃癌组与对照组比较TLR3c.1377基因型频率差异有统计学意义（P=0.025）,胃癌组T等位基因频率明显高于对照组（45.6% vs. 29.7%,P=0.004),T等位基因携带者的患病风险明显高于非携带者(OR=2.435,P=0.008）;（2）EBVaGC、EBVnGC以及对照组间TLR3c.1377基因型、等位基因频率差异无统计学意义（P>0.05）。（3）EBVaGC组,EBVnGC组以及正常对照组所有个体TLR4 Asp299Gly基因型均为Asp/Asp纯合子（P>0.05）。结论 TLR3c.1377基因多态性与胃癌易感性有关,T等位基因为胃癌的危险因子,而C等位基因为一保护基因;TLR3c.1377基因多态性与EBVaGC易感性无明显相关。     

基于DWI-ADC值动态监测宫颈癌放化疗疗效的临床研究

目的:通过DWI-ADC值测定动态监测宫颈癌放化疗疗效。方法:对2014年4月至2016年9月我院病理确诊的37例中晚期（FIGO分期Ⅱb-Ⅲb）宫颈鳞癌患者分别于同步放化疗前（A节点）、外照射治疗结束时（B节点）、后装内照射治疗3次时（C节点）、同步放化疗治疗结束时（D节点)行4次常规MRI及DWI检查。在ADC图上绘制ROI测量病灶、正常子宫肌层的ADC值;计算不同时间节点病灶的体积。所有数据经过统计学处理。结果:A、B、C、D时间节点病灶ADC值组间比较有明显差异(P<0.01),从A→D节点病灶ADC值逐步上升,其中ADC值上升最明显的为B节点,增幅32.55%;病灶体积缩小最明显为B-C节点,缩小率为65.64%。D节点病灶与A节点正常子宫肌层的ADC值比较无统计学差异。结论:基于ADC值测定能够更早、更精确、定量地动态评估中晚期宫颈鳞癌同步放化疗效果,并对预后疗效给予准确预测。     

胞苷脱氨酶基因对小鼠大剂量化疗的保护作用

Objective:To explore the feasibility of transfecting cytidine deaminase(CD)gene into mouse bone marrow cells in order to observe the drug resistance of high dose Ara-C and improve the tolerance of myelosuppression following combination chemotherapy.Methods:Human cytidine deaminase gene was transfected into mice bone marrow cells by retroviral vector.Resistant colony-forming unit granulocyte-macrophage(CFU-GM)assay was performed after the transfected mice bone marrow cells treated by the Ara-C.DNA was extracted from mice bone marrow cells.The drug resistant gene in mice bone marrow cells after transfection was detected by PCR.Results:Bone marrow cells of lhe donor mice cultured with lhe retroviral producer cells showed the drug resistant colonies and resistance to Ara-C,so did accept mice transplanted with the CD gene(CFU-GM of donor mice was 52%,X2=124.62,P＜0.01:accept mice was 54%,X2=126.26.P＜0.01,both compared with the contrast group).The animal survival rate was significantly higher in gene transfected group than that of the control(X2=7.42.P＜0.01).CD gene of transfected bone marrow cells was confirmed by PCR.Conclusion:CD gene can be transfected into bone marrow cells of mice efficiently and increase the drug resistance to Ara-C.     

儿童完全性重复肾伴输尿管囊肿的手术方式选择及疗效分析

目的：探讨儿童完全性重复肾伴输尿管囊肿的手术治疗方式,并分析其手术疗效。方法：回顾分析2012年1月至2018年7月行手术治疗的60例完全性重复肾伴输尿管囊肿患儿临床资料。其中,因上肾段发育不良行重复肾及输尿管切除术34例;上肾段发育良好者中,合并重度膀胱输尿管反流（vesicoureteral reflux,VUR）或非囊肿所在输尿管末端狭窄5例,行输尿管再植术,余21例行经尿道膀胱镜下输尿管囊肿开窗引流术。通过临床表现、肾积水程度、输尿管扩张程度、输尿管囊肿大小、VUR、再手术率等指标评估手术疗效。结果：术后60例患儿均获随访,随访时间1~55个月,平均11.6个月。行重复肾及输尿管切除术34例中,术后尿路感染5例;再行输尿管囊肿开窗引流2例,余32例中,B超仍可见输尿管囊肿6例,囊肿直径与术前相比有显著性差异[（13.2±4.4） mm vs. （26.3±3.9） mm,P=0.004]。行经尿道膀胱镜下输尿管囊肿开窗引流术21例中,术后尿路感染6例;再行重复肾及输尿管切除3例,再行输尿管膀胱再植术1例,余17例术后上肾段积水较术前减轻,差异有统计学意义[（9.9±8.2） mm vs. （24.9±10.8） mm,P=0.000],其中8例仍有输尿管扩张,较术前有统计学差异[（7.0±2.5） mm vs. （10.0±3.3） mm,P=0.007]。行输尿管膀胱再植术5例中,术后尿路感染1例,无再手术者,上肾段积水较术前减轻,差异有统计学意义[（10.0±2.9） mm vs. （24.2±6.9） mm,P=0.004]。结论：膀胱镜下输尿管囊肿开窗、重复肾及输尿管切除和输尿管再植术均可作为完全性重复肾伴输尿管囊肿的治疗方式。对于上肾段发育不良者,建议行上肾段及输尿管切除术。若上肾段发育良好,且合并重度膀胱输尿管反流或其他输尿管末端畸形者,建议行输尿管再植术;而无其他合并畸形者,可选择创伤小的经尿道膀胱镜下输尿管囊肿开窗术。     

靶向EGR1对泌乳素瘤的潜在治疗作用

目的：从基因分子水平出发,探究泌乳素瘤（prolactinoma,PRL）患者差异表达基因,从而探究疾病发展的核心驱动基因。旨在为泌乳素瘤患者的临床诊断、治疗及预后提供潜在的靶标。同时针对核心驱动基因,预测治疗泌乳素瘤患者的分子靶向药物。方法：从GEO（gene expression omnibus）数据库获取泌乳素瘤患者基因表达谱数据,进行泌乳素瘤组织和正常组织差异表达基因（differential expressed genes,DEG）筛选。同时进行加权基因共表达网络分析（weighted gene co-expression network analysis,WGCNA）、蛋白质相互作用网络分析（protein-protein interation,PPI）,确定泌乳素瘤发生发展的核心驱动基因。同时进行GO（gene ontology）分析、KEGG（kyoto encyclopedia of genes and genomes）分析及GSEA（gene set enrichment analysis）分析,探究泌乳素瘤患者发生改变的生物功能和信号通路,探究肿瘤发生发展的分子机制。此外,进行体外划痕实验、克隆形成测定、CCK-8实验及流式细胞术实验,验证分子靶向药物的潜在治疗效果。结果：共鉴定出1 201个差异表达基因,其中EGR1、MAPK1、MYC、BCL2和CALM1居于重要地位;GO和KEGG分析结果表明,泌乳素瘤的形成主要与纺锤极发生改变、卵母细胞减数分裂受到影响相关;泌乳素瘤很可能与帕金森病存在一定的同源性。同时进行CCK-8测定、克隆形成测定和体外划痕测定,验证了EGR1激动剂Genipin具有潜在抗泌乳素瘤的作用。流式细胞术分析结果显示,随着Genipin药物剂量增加,肿瘤细胞的凋亡百分比增加。结论：EGR1、MAPK1、MYC、BCL2和CALM1是泌乳素瘤的核心驱动基因,对泌乳素瘤的发生发展具有重要影响。EGR1的激动剂Genipin能够在体外抑制泌乳素瘤细胞的增殖和迁移,Genipin对泌乳素瘤具有潜在治疗作用。同时,本研究筛选出了泌乳素瘤发生的高风险信号通路,为其临床诊治提供了新的靶标,具有重要意义。