非小细胞肺癌中p-mTOR、Beclin1及Atg1蛋白表达及其临床意义

目的检测自噬相关蛋白p-mTOR、Beclin1及Atg1在非小细胞肺癌及正常肺组织中表达水平,探讨细胞自噬在肺癌演变过程中的作用机制。 方法应用免疫组织化学法测定96例非小细胞肺癌、64例正常肺泡、60例正常支气管断端组织p-mTOR、Beclin1及Atg1蛋白表达水平,分析其表达差异与临床病理特征相关性。 结果①Beclin1、Atg1蛋白在非小细胞肺癌组织中表达显著下降,且二者表达水平正性相关,p-mTOR蛋白表达显著升高;②随肿瘤增大、分化程度下降及出现淋巴结转移,Beclin1蛋白表达显著下降;③随肿瘤增大及出现淋巴结转移,Atg1蛋白表达显著下降;④随肿瘤分化程度下降及出现淋巴结转移,p-mTOR蛋白表达显著升高;⑤p-mTOR与Beclin1及与Atg1表达均呈负性相关。 结论自噬在非小细胞肺癌发病中起重要作用,自噬相关蛋白Beclin1、Atg1起协同抑癌作用,p-mTOR蛋白促进非小细胞肺癌发生、发展。     

白藜芦醇保护脑缺血/再灌注损伤与自噬关系的研究

目的研究白藜芦醇(resveratrol,Res)预处理对大鼠局灶性脑缺血/再灌注损伤后自噬的影响,探讨Res保护局灶性脑缺血/再灌注损伤的作用机制。方法将64只SD♂大鼠随机分成4组,假手术组(S组)、模型组(M组)、Res低剂量(15 mg.kg-1)干预组(R1组)和Res高剂量(40 mg.kg-1)干预组(R2组),用线栓法制备大鼠大脑中动脉栓塞(MCAO)模型,缺血2 h,再灌注24 h后,观察大鼠神经功能评分;透射电镜法观察神经元的超微结构及其内自噬空泡数量的变化;免疫组化法检测LC3的蛋白表达;RT-PCR法分别检测LC3和Beclin 1的mRNA表达。结果与M组相比,Res可明显减轻大鼠脑缺血/再灌注损伤的神经功能评分(P<0.05);使自噬泡数量明显增加;上调缺血皮质区脑组织LC3和Beclin 1蛋白及mRNA的表达(P<0.05)。结论 Res对大鼠局灶性脑缺血/再灌注损伤后神经元有保护作用,其机制可能与其诱导自噬的生成,上调LC3和Beclin 1的表达有关。     

Beclin1、LC3和mTOR在肝癌中的表达及临床意义

目的 通过检测Beclin1、LC3和雷帕霉素靶蛋白(mTOR)在肝癌组织中的表达,并结合各相关临床病理指标,从而探讨Beclin1、LC3和mTOR的表达水平在肝癌发生发展中的作用及其相互之间的关系.方法 采用免疫组织化学技术,检测56例肝癌组织和40例正常肝脏组织中Beclin1、LC3和mTOR蛋白的表达情况,并分析二者与肝癌各相关临床病理特征之间的关系.结果 Beclin1、LC3和mTOR蛋白在肝癌组织中的阳性表达率分别为75％、70％和50％,均明显高于正常肝组织中的阳性表达率25％、30％、28％,差异有统计学意义(P<0.05).Beclin1与LC3在肝癌中表达呈正相关(r=0.643,P<0.01),与mTOR表达之间无明显相关性(r=-0.167,P>0.05).LC3与mTOR表达呈负相关(r=-0.386,P<0.01).Beclin1、LC3和mTOR的表达均与年龄、性别、肿瘤大小、HBSAg、AFP值等均无显著相关性(P>0.05),但与病理学分级和有无胆管癌栓有关(P<0.05).结论 Beclin1、LC3和mTOR的异常高表达可能与肝癌的发生关系密切.     

p53抑癌基因在腺样囊性癌中的作用

腺样囊性癌是头颈部较常见的恶性肿瘤之一,其特点为嗜神经性及远处转移,手术和放化疗效果及预后较差,为了提高患者的远期生存率,迫切需要寻找新的治疗方法,而p53基因是一种重要的抑癌基因,近年来在恶性肿瘤中的作用日益受到重视,因此可作腺样囊性癌治疗的重要途径来探讨。     

Curcumin inhibits pancreatic cancer cell proliferation by regulating Beclin1 expression and inhibiting the hypoxia-inducible factor-1α-mediated glycolytic pathway

BackgroundPancreatic cancer has a high degree of malignancy and high mortality. Understanding its biological status can provide more therapeutic targets for the future. The present study was to investigate whether curcumin can inhibit pancreatic cancer cell proliferation by regulating Beclin1 expression and inhibiting the hypoxia-inducible factor-1α (HIF-1α)-mediated glycolytic pathway.MethodsTwo pancreatic cancer cell lines, PANC-1 and SW1990, were treated with different concentrations of curcumin (0, 20, 40, and 60 µM). Cell viability was detected using the Cell Counting Kit-8 (CCK-8) assay and flow cytometry was performed to determine the apoptosis rate and cell cycle arrest of the pancreatic cancer cells. PANC-1 and SW1990 cells were treated with different concentrations of curcumin under hypoxic conditions for 48 hours to detect the relative expression of the Beclin1 protein. The co-immunoprecipitation (co-IP) method was used to determine whether curcumin could inhibit the interaction between Beclin1 and HIF-1α.ResultsThe proliferation inhibition rates of PANC-1 cells after exposure to 0, 20, 40, and 60 µM curcumin were 0%, 31.6%, 47.2%, and 63.9%, respectively, and that of SW1990 cells were 0%, 18.8%, 46.3%, and 63.5% respectively. Western blot analyses showed decreased expression of Beclin1 in cells treated with curcumin. The expression of Beclin1 in the nucleus and cytoplasm decreased with increasing concentrations of curcumin. Co-IP results demonstrated that curcumin inhibited the interaction between Beclin1 and HIF-1α. Treatment with the higher doses of curcumin (40 and 60 µM) significantly decreased the protein expression levels of HIF-1α. In addition, the expression levels of Kidney-Specific Cadherin (HSP70, HSP90, and von Hippel-Lindau protein (pVHL) were significantly decreased in pancreatic cancer cells while the expression of prolyl hydroxylase (PHD) and receptor of activated protein kinase C (RACK1) increased significantly. Furthermore, curcumin reduced cellular adenosine triphosphate (ATP) production in a dose-dependent manner. Compared with control pancreatic cancer cells, the expression levels of GLUT1, HK2, LDHA, and PDK1 gradually decreased with increasing curcumin concentrations.ConclusionsCurcumin can inhibit the expression of Beclin1 and HIF-1α in pancreatic cancer cells under anoxic conditions, thereby affecting the glycolysis pathway and inhibiting cell proliferation.     

慢加急性乙型肝炎肝衰竭患者血清自噬相关蛋白水平及其临床意义探讨

目的 探讨慢加急性乙型肝炎肝衰竭(HBV-ACLF)患者血清p62、微管相关蛋白1轻链3-Ⅱ(LC3-Ⅱ)和Beclin1水平变化及其临床意义.方法 2018年9月～2020年9月我院收治的HBV-ACLF患者60例和同期体检的健康人65例,计算终末期肝病模型评分(MELD),采用ELISA法测定血清p62、Becli...     

TAp73和Beclin1在人骨肉瘤中表达及其临床意义

目的检测抑癌基因TAp73与自噬相关因子Beclin1在人骨肉瘤中的表达,探讨TAp73与肿瘤自噬的关系。方法采用免疫组织化学SP法检测TAp73、Beclin1在48例骨肉瘤患者的瘤体组织中的表达,结合患者的临床资料进行相关性分析。结果 TAp73在人骨肉瘤组织中的阳性率54.17%,Beclin1的阳性率为62.50%,TAp73阳性表达,其Beclin1表达程度较高(Z=-2.146,P=0.032)。在人骨肉瘤中TAp73的表达程度与Beclin1的表达程度呈正相关(r=0.470,P=0.001)。TAp73、Beclin1阳性表达的患者总体生存率优于阴性表达(80.8%vs 50.0%,P=0.000;80.0%vs 44.4%,P=0.001)。TAp73、Beclin1不同表达程度与患者的生存时间呈正相关(r=0.696,P=0.00;r=0.372,P=0.009)。结论在人骨肉瘤中TAp73可能与骨肉瘤中Beclin1依赖的肿瘤自噬活性上调有关。检测TAp73和Beclin1在人骨肉瘤中的表达水平有助于患者临床预后判断。     

琥珀散对人MenSCs植入式子宫内膜异位症裸鼠异位病灶及Beclin1、ZEB1表达的影响

目的 探究琥珀散对人经血源间充质干细胞(MenSCs)植入式子宫内膜异位症(EMT)裸鼠的异位病灶及 Bcl-2 同源结构域蛋白抗体(Beclin1)、锌指E盒同源结合蛋白1(ZEB1)表达的影响。方法 将裸鼠随机分为对照组、模型组、地诺孕素(0.001 g·kg^-1)组和琥珀散(29 g·kg^-1)组,每组 10 只。从 EMT 患者及健康女性的月经血中提取 MenSCs,并借助成脂、成骨诱导分化鉴定其干细胞属性。将EMT MenSCs注射至裸鼠腹部皮下,制备EMT裸鼠模型,对照组植入正常人MenSCs。至第7天每组随机抽取1只裸鼠,开腹观察异位病灶生长情况,并借助HE染色和人类白细胞抗原A(HLAA)免疫荧光法评价模型。自造模成功连续给药21 d,末次给药后第2日取材,观测EMT异位病灶体积及血管分布;切取异位病灶组织,应用实时荧光定量PCR(qRT-PCR)、Western blotting、免疫组织化学及免疫荧光技术分别测定组织中Beclin1与ZEB1在mRNA及蛋白水平的表达。结果 EMT源及正常MenSCs为长梭形,呈辐射状向周边集落扩散,且经成脂、成骨诱导分化后均有脂滴、钙结节形成。造模7 d后,对照组未见异位病灶形成,其余各组均有囊泡样病灶形成,且周围血管形成丰富,病理切片均可见子宫内膜腺体和间质,HLAA 均阳性表达。与模型组比较,琥珀散能显著缩小EMT裸鼠异位病灶体积(P<0.001),改善周围血管分布。在 mRNA水平, 与模型组比较,地诺孕素组和琥珀散组ZEB1表达显著降低(P<0.001),Beclin1 表达显著升高(P<0.001)。在蛋白水平,与模型组比较,Western blotting结果显示,地诺孕素组和琥珀散组ZEB1 表达显著降低 (P<0.05),Beclin1表达显著升高(P<0.01、0.001);免疫组化结果显示,琥珀散组 Beclin1 大量分布在异位内膜、腺腔以及腺上皮细胞与间质细胞的胞核及胞浆中,Beclin1 表达显著增加(P<0.01);免疫荧光结果显示,琥珀散组 ZEB1 蛋白在异位内膜、腺腔、腺上皮细胞及周围间质细胞的胞核与胞浆中的分布均明显减少,地诺孕素组、琥珀散组 ZEB1 蛋白表达阳性率显著降低(P<0.001)。结论 琥珀散可能通过上调 Beclin1、下调 ZEB1 表达,缩小 EMT 裸鼠异位病灶,改善周围血管分布。     

The Suppressive Effects of the Petroleum Ether Fraction from Atractylodes lancea (Thunb.) DC. On a Collagen‐Induced Arthritis Model

In Chinese traditional medicine, the rhizome of Atractylodes lancea (Thunb.) DC. (A. lancea) is used extensively for the treatment of several diseases such as rheumatic diseases, but its actions on rheumatoid arthritis have not been clarified. The purpose of this article was to investigate the pharmacological effect of an A. lancea rhizome extract on collagen‐induced arthritis (CIA) in rats. The CIA model was induced by the injection of bovine type II collagen. The rats were orally administered the petroleum ether (PE) fraction of the A. lancea rhizome (0.82 and 1.64 mg/kg), methotrexate (0.3 mg/kg body weight), or a vehicle from day 7 to day 15 after the model was established. The histological examination and radiological observation showed that the PE fraction significantly reduced the inflammatory responses and collagen loss in the joints of the rats with CIA. The PE fraction inhibited the production of tumor necrosis factor‐α, interleukin (IL)‐1β, IL‐17, and IL‐6 in the sera. Moreover, the treatment with the PE fraction in vivo was able to reduce the level of Beclin 1 protein in the synovial tissue of the rats. These results highlight the antiarthritic potential of the PE fraction of the A. lancea rhizome and provide further evidence of the involvement of Beclin 1 inhibition in the effects of the PE fraction of the A. lancea rhizome. Copyright © 2016 John Wiley & Sons, Ltd.     

Novel pharmacological modulators of autophagy: an updated patent review (2012-2015)

Introduction: Autophagy is a lysosome-dependent degradation pathway that maintains cellular homeostasis in response to a variety of cellular stresses. Accumulating reports based on animal models have indicated the importance of this catabolic program in many human pathophysiological conditions, including diabetes, neurodegenerative diseases, aging, and cancers. Therefore, autophagy has been highlighted as a novel therapeutic target with a wide range of beneficial effects on human diseases. Here, we review the recent advances of our knowledge toward autophagy, as well as the efforts for developing autophagy modulators.

Areas covered: The relevant patents (published at 2012–2015) and the research literature claiming the pharmacological modulation of autophagy are reviewed. Also, their molecular mechanisms and potential therapeutic utilities are discussed.

Expert opinion: Considering the molecular machinery involved in autophagy induction, the targeting of autophagy-specific protein is very important to design the therapeutic interventions for specifically treating a variety of autophagy-associated disorders. Many patents and the research literature described in this review have shown promising applications of the relevant autophagy modulators for cancer or neurodegeneration treatments, a few of which are already being considered for clinical evaluation. However, most patents have claimed the modulators of autophagy with little information regarding their mechanisms of action. To design highly potent therapeutics, further work, such as developing compounds that specifically target the autophagy-specific machinery, are required.