热应激作用下人肺腺癌A549细胞中HSP70B’的动态表达

[目的]探讨热休克蛋白70B’（HSP70B’）的产生条件，分析HSP70B’在人肺腺癌A549细胞中的表达特征。[方法]采用Western-blot方法检测正常培养细胞（37℃）和不同温度（38、39、40、41、42、43、44、45℃）下热应激1h；选择HSP70B’表达最高的温度42℃，应激1h后恢复培养（37℃，5％CO2）不同时间（0、3、6、9、12、15、18、24h）；在42℃应激不同时间（0、20、40、60、80、100、120、140、180min）恢复培养6h的肺腺癌A549细胞中HSP70B’蛋白的表达水平。[结果]HSP70B’在正常状态的A549细胞中不表达；在不同应激温度作用下，HSP70B’在39℃开始表达，在42℃时表达量达到最高水平；细胞维持42℃应激后恢复培养不同时间作用下，HSP70B’表达在恢复培养6h后达最高，恢复培养24h后降到最低水平；在42℃应激不同时间后均恢复培养6h作用下，热应激60min时HSP70B’表达最高。[结论]肺腺癌细胞株A549细胞中HSP70B’产生的最优条件是42℃应激60min后37℃恢复培养6h。HSP70B’在热应激后的A549细胞中迅速表达，其在细胞热应激中的作用及意义有待进一步研究。     

绿原酸诱导肺癌细胞凋亡及其机制研究

目的研究绿原酸对人肺癌A549细胞凋亡的作用及其机制,为临床应用提供依据。方法四氮唑盐(MTT)法检测不同浓度的绿原酸对A549细胞增殖的影响;流式细胞术检测细胞凋亡率;采用间接荧光标记法测定细胞内活性氧水平的变化;分光光度法检测绿原酸对表皮生长因子受体酪氨酸激酶(epidermal growth factor receptor tyrosine kinase,EGFR-TPK)、基质金属蛋白酶(matrix metalloproteinase 2,MMP-2)和氨肽酶N(aminopeptidase,APN)活性的变化。结果MTT法检测显示,绿原酸在24 h、48 h和72 h时均可抑制A549细胞增殖,且呈浓度和时间依赖性关系趋势;绿原酸作用48 h后,细胞凋亡率呈现明显增高的趋势,与对照组比较差异有显著性(P0.01);在100μg/ml质量浓度的绿原酸作用下使细胞内活性氧(ROS)百分率达到了(42.32±3.3)%,与对照组相比比较差异有显著性(P0.01);绿原酸对APN、EGFR-TPK和MMP-2半数抑制浓度(IC_(50))分别为(8.12±0.78)、(29.43±2.76)和(1.54±0.13)μg/ml,显示出了较好的抑制效果。结论绿原酸可以抑制A549的增殖和转移,促进细胞凋亡,其凋亡机制可能与细胞内活性氧组分的增多及APN、EGFR-TPK和MMP-2活性降低有关。     

Nec-1对铝作用下神经细胞活力及凋亡、自吞噬基因的影响

[目的]研究坏死性抑制剂Necrostatin-1（Nec-1）对铝（Al）作用下的神经细胞活力及凋亡、自吞噬基因的影响。[方法]体外原代培养小鼠神经细胞,通过2mmol／LAl3＋作用于神经细胞制造铝损伤模型,加入不同剂量的Nec-1,用细胞活力检测（CCK-8）和逆转录．聚合酶链反应（RT．PCR）的方法观察Nec-1对染铝神经细胞的作用。[结果]细胞活力检测：以2mmol／LAl3＋染毒组作为对照,30μmol／LNec-1组的细胞活力与对照组比较,差别有统计学意义（P〈0．05）,60、90μmol／LNec-1组与对照组比较,差异均有统计学意义（P〈0．01）.RT-PCR结果：2mmol／LAl3＋染毒组caspase-3基因的表达为0．98±0．120,而Nec-160μmol／L组和Nec-190μmol／L组caspase-3的表达分别为0．50±0．077和0．44±0．050,同对照组相比,两组caspase-3的表达均下降（P〈0．01）。2mmol/LAl3＋染毒组的LC3-Ⅱ基因的表达为1,82±0．047,而60μmol／LNec-1组和90μmol／LNec-1组的LC3．口表达分别为1．00±0．022和0．97±0．035,同对照组相比,两组LC3-Ⅱ的表达均呈下降（P〈0．01）。[结论]Nec-1可使铝作用下的神经细胞活力上升,并使神经细胞的自吞噬和凋亡基因表达下降。     

放化疗同步与序贯治疗局部晚期非小细胞肺癌临床疗效比较

目的比较紫杉醇联合顺铂同步放疗及序贯放化疗治疗局部晚期非小细胞肺癌的生存期、疗效和不良反应。方法 78例NSCLC病例随机分为同步放化疗组39例（治疗组）和序贯放化疗组39例（对照组）。比较两组疗效和不良反应。结果治疗组的近期疗效和生存率均优于对照组,差异有统计学意义（P〈0.05）。两组均无明显心、肝肾功能损害,无治疗相关性死亡发生。两组不良反应主要表现为放射性食管炎、恶心呕吐、骨髓抑制、放射性肺炎,不良反应差异均无统计学意义（P〉0.05）,经对症治疗后均耐受。结论同步放化疗联合治疗局部晚期NSCLC可提高局部控制率延长生存期,改善生活质量,可耐受。     

临床路径在肺癌放化疗中的应用及成效

介绍了肺癌放化疗临床路径的建立及实施。结果显示,应用临床路径可以有效缩短肺癌放化疗的平均住院日,降低平均费用,提高病人满意度,减少医疗纠纷。     

JNK信号通路在二甲基肿酸所致人胚肺成纤维细胞DNA损伤与凋亡中的作用

[目的]探讨c-Jun氨基末端激酶（JNK）信号通路在二甲基胂酸（DMA）所致人胚肺成纤维（HELF）细胞DNA损伤与凋亡中的作用。[方法]以0、2．5、5、10和20μmol／LDMA处理HELF细胞48h后，检测HELF细胞生长、DNA损伤、细胞凋亡、JNK磷酸化水平；并用20μmol／LJNK抑制剂SP600125预处理30min后，再用DMA处理HELF细胞48h，观察上述指标变化情况。[结果]10和20μmol／LDMA对HELF细胞生长产生明显抑制作用（P〈0．05）；2．5、5、10和20μmol／LDMA引起HELF细胞γ-H2AX表达水平明显增强（P〈0．05），并具有一定剂量-效应关系（经直线相关分析，r=-0．982，P〈0．05）；5、10和20μmol／LDMA引起HELF细胞断裂，caspase3表达水平明显增强（P〈0．05），呈一定剂量-效应关系（经直线相关分析，r=0．945，P〈0．05）；5、10和20μmol／LDMA处理HELF细胞48h后，JNK磷酸化的表达水平明显增加（P〈0．05），呈现一定剂量-效应关系（经直线相关分析，r=0．988，P〈0．05）；JNK抑制剂SP600125能明显降低10、20μmol／LDMA的生长抑制作用（P〈0．05）；JNK抑制剂SP600125可明显阻滞DMA所致HELF细胞对DNA损伤和诱导的凋亡作用（P〈0．05）。[结论]DMA可引起HELF细胞DNA损伤和凋亡；在此过程中JNK信号通路激活起到正调控作用。     

青石棉诱导A549细胞ERK1／2及E1k1激活的研究

目的　研究细胞外信号调节蛋白在青石棉致肺部疾病的作用。方法　用Western免疫印迹、免疫沉淀等对石棉刺激A5 4 9细胞后细胞外信号调节蛋白及其下游Elk1蛋白磷酸化等进行了研究。结果　石棉刺激细胞后 ,磷酸化ERK1 2、Elk1等高表达 ,差异有显著性 (P <0 0 5 )。结论　ERK1 2、Elk1磷酸化可能参与青石棉的致病过程     

青石棉诱导A549细胞ERK1/2及Elk1激活的研究

目的　研究细胞外信号调节蛋白在青石棉致肺部疾病的作用。方法　用Western免疫印迹、免疫沉淀等对石棉刺激A5 4 9细胞后细胞外信号调节蛋白及其下游Elk1蛋白磷酸化等进行了研究。结果　石棉刺激细胞后 ,磷酸化ERK1 2、Elk1等高表达 ,差异有显著性 (P <0 0 5 )。结论　ERK1 2、Elk1磷酸化可能参与青石棉的致病过程     

组织蛋白酶B与紫杉醇诱导胃癌BGC-823细胞凋亡的体外实验研究

目的 通过体外实验研究半胱氨酸蛋白酶B(Cathepsin B)与紫杉醇诱导胃癌BGC-823细胞凋亡的关系.方法 将BGC-823细胞(密度为1×105/孔)分别暴露于紫杉醇溶液(终浓度为0～0.5μmol/L)及紫杉醇(终浓度为0.3 μmol/L)+Cathepsin B特异性抑制剂[CA-074(Me),终浓度...     

青蒿琥酯联合热疗诱导肺腺癌A549细胞凋亡作用

目的 探讨青蒿琥酯联合热疗诱导肺腺癌A549细胞凋亡的线粒体途径作用机制。方法 150μmol/L青蒿琥酯、青蒿琥酯联合热疗(43℃,1h)作用A549细胞24h,流式细胞技术检测细胞周期和凋亡率情况。荧光显微镜观察线粒体膜电位变化。免疫组化、western blot检测细胞色素c、Bcl-2和caspase-3表达,CaspACETM检测试剂盒检测活性caspase-3表达。结果 青蒿琥酯、青蒿琥酯联合热疗作用A549细胞24h后,G0/G1期细胞数增多,S期和G2/M期细胞数减少(P<0.01);凋亡率分别是16.77%和28.90%(P<0.01)。线粒体膜电位下降率分别为18.05%,50.98%(P<0.01)。细胞色素c蛋白、caspase-3表达增加而Bcl-2蛋白表达减少(P<0.01)。活性caspase-3蛋白水平增加A值分别为0.2641±0.0027,0.3613±0.0019,与对照组相比,差异有统计学意义(P<0.01)。结论 青蒿琥酯、青蒿琥酯联合热疗可诱导肺腺癌A549细胞凋亡,激活线粒体途径可能是诱导凋亡重要作用机制。     

环境雌激素对乳腺癌细胞株MCF-7凋亡作用的影响

[目的] 通过观察对-壬基酚(nonylphenol,NP)、双酚A(bisphenol A,BisA)和邻苯二甲酸二丁酯(dibutylphthalate,DBP)对乳腺癌细胞株MCF-7凋亡的影响,探讨环境雌激素促进MCF-7细胞增殖的生物学机制.[方法] 将在DMEM培养液(含10%小牛血清)中的MCF-7细胞采用开放式单层贴壁培养,加受试物前5 d将培养细胞用PBS洗涤后改为在无酚红DMEM(含5%经活性碳-葡聚糖苷处理的胎牛血清,CDT-FBS)中培养连续5 d,其目的是耗尽细胞内储存的雌激素.实验设溶剂对照组、雌激素对照组、抗雌激素(它莫西芬)对照组及三个实验组,采用流式细胞术、DNA片段凝胶电泳和细胞苏木素染色(HE),观察环境雌激素对雌激素耗尽所诱导的细胞凋亡作用的影响.[结果] 3.2×10-6 mol/L NP和3.2×10-5 mol/L BisA对MCF-7细胞处理72 h,与对照组相比,流式细胞仪分析结果表明,二倍体细胞(G1期细胞)明显增加,亚二倍体细胞峰(即细胞凋亡率)消失,DNA凋亡片段凝胶电泳不显示典型的凋亡DNA片段梯带,细胞苏木素染色结果也显示凋亡细胞明显减少.[结论] 与溶剂对照组相比,NP和BisA均能够抑制MCF-7细胞的凋亡作用,而3.2×10-4 mol/L DBP对细胞凋亡的影响作用不明显.这一结果提示,该类物质有可能是通过抑制细胞凋亡而发挥其促进MCF-7细胞增殖作用的.     

沙利度胺联合顺铂对A549肺腺癌细胞株的体外实验

目的观察沙利度胺与顺铂（DDP）联合应用对A549人肺腺癌细胞凋亡的影响,观察沙利度胺对肺腺癌A549细胞VEGF、BFGF蛋白表达的影响。方法采用PI单染法流式细胞检测细胞凋亡率;采用免疫细胞化学方法检测沙利度胺处理后肺腺癌A549细胞VEGF、BFGF蛋白表达的变化。结果在一定浓度范围内,沙利度胺联合DDP处理组细胞凋亡率明显高于单药组;沙利度胺（7.5mg/L）作用于人肺腺癌A549细胞后,处理组的VEGF、BFGF蛋白表达与未加药组比较差异显著。结论沙利度胺DDP联合,可增加A549人肺腺癌细胞的凋亡率;沙利度胺可抑制A549人肺腺癌细胞VEGF、BFGF的蛋白表达。     

Impact of Genetic Polymorphisms on the Metabolic Pathway of Vitamin D and Survival in Non-Small Cell Lung Cancer

Vitamin D has been associated with risk, development, and progression of cancer. However, the genes involved in its metabolism are highly polymorphic, compromising its activity. The aim of this study is to evaluate the association between the gene polymorphisms involved in the metabolic pathway of vitamin D and survival in patients with non-small-cell lung cancer (NSCLC). The study was designed as an observational cohort which included 194 Caucasians patients from southern Spain with NSCLC. Real-time polymerase chain reaction was used to analyze the following polymorphisms: CYP27B1 rs4646536, rs3782130, and rs10877012; CYP24A1 rs6068816 and rs4809957; GC rs7041; CYP2R1 rs10741657; VDR rs1544410 (BsmI), rs11568820 (Cdx-2), rs2228570 (FokI), rs7975232 (ApaI), and rs731236 (TaqI). Progression-free survival (PFS) and overall survival were assessed. Cox regression showed that rs4646536 was associated with PFS in the general population (p = 0.0233) and in the non-resected NSCLC subgroup (p = 0.0233). In the resected NSCLC subgroup, rs11568820 was associated with OS (p = 0.0129) and rs7041 with PFS (p = 0.0447). In the non-resected NSCLC subgroup, rs6068816 was associated with PFS (p = 0.0048) and OS (p = 0.0089) and rs731236 and rs7975232 were associated with OS (p = 0.0005) and PFS (p = 0.0002), respectively. The other polymorphisms showed no effect on the results. The rs4646536, rs6068816, rs7041, rs11568820, rs731236, and rs7975232 polymorphisms are associated with survival in NSCLC and may have a substantial role as prognostic markers of the disease.     

Resveratrol against Cervical Cancer: Evidence from In Vitro and In Vivo Studies

Cervical cancer affects many women worldwide, with more than 500,000 cases diagnosed and approximately 300,000 deaths each year. Resveratrol is a natural substance of the class of phytoalexins with a basic structure of stilbenes and has recently drawn scientific attention due to its anticancer properties. The purpose of this review is to examine the effectiveness of resveratrol against cervical cancer. All available in vitro and in vivo studies on cervical cancer were critically reviewed. Many studies utilizing cervical cancer cells in culture reported a reduction in proliferation, cell cycle arrest, and induction of apoptosis. Apart from apoptosis, induction of autophagy was seen in some studies. Importantly, many studies have shown a reduction in the HPV oncoproteins E6 and E7 and increased levels of the tumor suppressor p53 with resveratrol treatment. A few studies examined the effects of resveratrol administration in mice ectopic-xenografted with cervical cancer cells showing reduced tumor volume and weight. Overall, the scientific data show that resveratrol has the ability to target/inhibit certain signaling molecules (EGFR, VEGFR, PKC, JNK, ERK, NF-kB, and STAT3) involved in cervical cancer cell proliferation and survival. Further in vivo experiments and clinical studies are required to better understand the potential of resveratrol against cervical cancer.     

Hesperetin Induces Apoptosis in Human Glioblastoma Cells via p38 MAPK Activation

Abstract

Glioblastoma (GBM) is the most common and aggressive form of malignant brain tumor, with poor prognosis and a lack of effective treatment. Hesperetin, a natural product found in citrus fruits, displayed bioactivities including antioxidant, anti-inflammatory, and anticancer, while its effects on GBM cells were largely unknown. Here, we explored the anticancer effect of hesperetin on human GBM cells in vitro, as well as the underlying signaling mechanisms. By CCK-8 assay and live/dead assay, hesperetin presented significant inhibitory effect on human GBM U-251 and U-87 cell viability. By DAPI staining and Annexin V-FITC/PI assay, apoptotic death was proved to contribute to the cell viability reduction, and it was verified by the increased Bax/Bcl-2 ratio in western blotting results. Furthermore, by cell cycle analysis and western blotting for cyclin B1, CDK1, and p21, hesperetin was found to induce cell-cycle arrest at G2/M phase. For signaling mechanism, the western blotting results showed elevated p38 MAPK activation, and the reduced Bcl-2 and enhanced Bax upon hesperetin treatment were partly reversed by p38 MAPK inhibitor SB203580. In summary, we have discovered hesperetin as a natural product candidate for the treatment of GBM, and that it could induce GBM cell apoptosis via p38 MAPK activation.     

双酚A对人胚肝细胞凋亡的影响

[目的]研究双酚A(BPA)对人胚肝L-02细胞的凋亡的影响。[方法]以MTT来分析不同浓度BPA对人胚肝L-02细胞的细胞存活率的影响;用流式细胞术分析不同浓度BPA对细胞凋亡的影响;通过RT-PCR分析BPA引起的生长抑制、DNA损伤基因GADD45、GADD153、GADD34表达水平的变化。[结果]BPA处理后,L-02细胞的生存率随着BPA浓度的增加而下降,在≥50μmol/L,细胞的存活率显著降低(P<0.01)。随着BPA浓度升高,细胞凋亡率增加,≥50μmol/L可显著升高L-02细胞的凋亡率(P<0.01)。生长抑制DNA损伤基因GADD45、GADD153.GADD34表达水平随着BPA浓度增加表达量也增加,100μmol/LBPA组显著高于正常对照组,但在100μmol/L,BPA+VitC组三种GADD基因表达量均稍降低。[结论]BPA对L-02细胞具有细胞毒性作用,可导致细胞凋亡率升高,与细胞生长调控和DNA损伤相关的GADD45、GADD153、GADD34基因表达升高,VitC可减缓BPA对DNA的损伤作用。     

DC-CIK联合化疗治疗晚期非小细胞肺癌的近期临床疗效

目的：观察DC-CIK联合化疗治疗晚期非小细胞肺癌的近期疗效及不良反应。方法：纳入62例Ⅲ～Ⅳ晚期非小细胞肺癌患者,根据随机数字表将患者随机纳入治疗组和对照组。治疗组采用DC-CIK联合NP化疗方案;对照组采用单纯NP化疗。治疗两个疗程,比较两组患者治疗前后的瘤体大小变化、免疫功能和副反应。结果：治疗组瘤体变化有效率45.16%,与对照组的32.26%比较差异无统计学意义（P〉0.05）,治疗组瘤体稳定率83.87%,明显优于对照组的61.29%（P〈0.05）;治疗组治疗后外周血T细胞亚群无明显变化（P〉0.05）,对照组患者治疗后外周血CD3＋、CD4＋细胞比例明显下降（P〈0.05）;治疗组的不良反应与单纯化疗组比较差异无统计学意义（P〉0.05）。结论：与单纯化疗相比,应用DC-CIK联合化疗治疗中晚期非小细胞肺癌近期疗效较好,可有效控制瘤体,保护和提高患者细胞免疫功能。     

非小细胞肺癌中S100A4和上皮钙粘蛋白的表达及意义

目的 探讨非小细胞肺癌中S100A4和上皮钙黏蛋白(E-cad)的表达及意义.方法 采用免疫组化SP法检测76例乳腺癌中S100A4,E-cad表达情况,分析二者的表达与临床病理特征之间的关系.结果 S100A4的表达与组织学分级、淋巴结转移呈正相关(P＜0.01).E-cad的表达与组织学分级、淋巴结转移呈负相关(P＜0.01).结论 非小细胞肺癌中S100A4的表达和E-cad的表达缺失与肿瘤的进展密切相关,是判断非小细胞肺癌生物学行为、预测转移趋势的有价值的指标.     

Coffeeberry Activates the CaMKII/CREB/BDNF Pathway,Normalizes Autophagy and Apoptosis Signaling in Nonalcoholic Fatty Liver Rodent Model

Nonalcoholic fatty liver disease (NAFLD) shows extensive liver cell destruction with lipid accumulation, which is frequently accompanied by metabolic comorbidities and increases mortality. This study aimed to investigate the effects of coffeeberry (CB) on regulating the redox status, the CaMKII/CREB/BDNF pathway, autophagy, and apoptosis signaling by a NAFLD rodent model senescence-accelerated mice prone 8 (SAMP8). Three-month-old male SAMP8 mice were divided into a control group and three CB groups (50, 100, and 200 mg/kg BW), and fed for 12 weeks. The results show that CB reduced hepatic malondialdehyde and carbonyl protein levels. CB significantly enhanced Ca²⁺/calmodulin-dependent protein kinase II (CaMKII) and brain-derived neurotrophic factor (BDNF) and reduced the phospho-cAMP response element-binding protein (p-CREB)/CREB ratio. In addition, CB increased the silent information regulator T1 level, promoted Beclin 1 and microtubule-associated protein light chain 3 II expressions, and reduced phosphorylated mammalian target of rapamycin and its downstream p-p70s6k levels. CB also inhibited the expressions of apoptosis-related factors poly (ADP-ribose) polymerase-1 and the apoptosis-inducing factor. In conclusion, CB might protect the liver by reducing oxidative stress, activating the CaMKII/CREB/BDNF pathway, and improving autophagic and apoptotic expressions in a dose-dependent manner.     

顺铂对A549细胞DNA修复酶MGMT和DNA-PKcs表达的影响

[目的]探讨在DNA损伤剂顺铂作用下，A549细胞内DNA修复基因六氧甲基鸟嘌呤DNA甲基转移酶(MGMT)和DNA依赖的蛋白激酶复合物催化亚单位(DNA-PKcs)的表达变化。[方法]MTT法分析顺铂对A549细胞生存力的影响；彗星实验研究顺铂对DNA损伤的程度；RT-PCR分析不同顺铂作用时问修复酶mRNA水平的变化。[结果]顺铂对A549细胞的生长有明显的抑制作用，且为剂量依赖性。低浓度(IC20剂量)顺铂作用下，DNA损伤程度的变化与作用时间成正比，且伴随MGMT和DNA-PKcs的表达上升。停止作用后，DNA损伤程度减轻，修复基因的表达也逐渐恢复至正常水平。[结论]短期顺铂染毒所导致的急性DNA损伤对MGMT和DNA-PKcs的表达有明显的正性诱导作用。