鱼藤素通过调控miR-520a-3p影响卵巢癌SKOV3细胞的增殖和凋亡

目的：探讨鱼藤素通过调控miR-520a-3p表达对卵巢癌SKOV3细胞增殖和凋亡的影响。方法：将SKOV3细胞分为对照组（鱼藤素0μmol/L）、鱼藤素低剂量（5μmol/L）、中剂量（10μmol/L）、高剂量（20μmol/L）组,miR-NC组、过表达miR-520a-3p组,鱼藤素+anti-miR-NC组、鱼藤素+anti-miR-520a-3p组。CCK-8法、细胞集落形成实验、FCM以及qPCR法分别检测SKOV3细胞的增殖抑制率、细胞克隆形成数、凋亡率以及miR-520a-3p表达水平。结果：与对照组比较,鱼藤素（低、中、高剂量）组SKOV3细胞增殖抑制率、凋亡率、miR-520a-3p表达水平均显著升高（均P<0.05）,细胞克隆形成数显著减少（P<0.05）。与miR-NC组比较,过表达miR-520a-3p组SKOV3细胞的增殖抑制率、凋亡率均显著升高（均P<0.05）,细胞克隆形成数显著减少（P<0.05）。与鱼藤素+anti-miR-NC组比较,鱼藤素+anti-miR-520a-3p组SKOV3细胞的增殖抑制率、凋亡率均显著降低（均...     

lncRNA NEAT1调控miR-4262表达影响甲状腺癌细胞增殖、迁移和侵袭的机制研究

目的:研究长链非编码RNA NEAT1（lncRNA NEAT1）对甲状腺癌细胞增殖、迁移和侵袭的影响及其作用机制。方法:qPCR检测NEAT1和miR-4262在人正常甲状腺滤泡上皮细胞株Nthy-ori 3-1与甲状腺癌细胞株FTC-133、ML-1、SW579中的表达。用si-NEAT1或miR-4262 mimic转染SW579细胞,考察其在细胞增殖、迁移和侵袭中的作用。生物学信息预测结合双荧光素酶报告实验分析NEAT1和miR-4262之间的靶向关系。MTT法检测细胞增殖抑制率;Western blot检测细胞周期蛋白D1（CyclinD1）、p21、基质金属蛋白酶-2（MMP-2）和基质金属蛋白酶-9（MMP-9）表达;Transwell小室法检测细胞迁移和侵袭。共转染si-NEAT1和anti-miR-4262,观察抑制miR-4262表达对抑制NEAT1表达诱导的SW579细胞增殖、迁移及侵袭的影响。结果:NEAT1在甲状腺癌细胞中的表达显著上调（P＜0.05）,miR-4262表达下调（P＜0.05）。抑制NEAT1表达或过表达miR-4262显著增加SW579细胞增殖抑制率和p21蛋白表达量（P＜0.05）,显著降低迁移细胞数、侵袭细胞数及CyclinD1、MMP-2和MMP-9蛋白表达量（P＜0.05）。miR-4262是NEAT1的靶基因。上调或下调NEAT1表达明显调控miR-4262表达（P＜0.05）。抑制miR-4262表达能逆转抑制NEAT1表达对SW579细胞增殖抑制率和p21蛋白表达的促进作用,以及对细胞迁移、侵袭及CyclinD1、MMP-2和MMP-9蛋白表达的抑制作用。结论:lncRNA NEAT1通过靶向miR-4262影响甲状腺癌细胞增殖、迁移和侵袭。     

lncRNA LUADT1靶向miR-138-5p调控胶质瘤细胞的增殖、迁移和侵袭

目的:探讨肺腺癌相关转录本1（LUADT1）对胶质瘤细胞的增殖、迁移和侵袭的影响及分子机制。方法:将si-NC、si-LUADT1、miR-NC、miR-138-5p、pcDNA、pcDNA-LUADT1分别转染至U251细胞中,记为si-NC组、si-LUADT1组、miR-NC组、miR-138-5p组、pcDNA组、pcDNA-LUADT1组;将si-LUADT1分别与anti-miR-NC、anti-miR-138-5p共转染至U251细胞中,记为si-LUADT1+anti-miR-NC组、si-LUADT1+anti-miR-138-5p组。实时荧光定量PCR（RT-qPCR）检测LUADT1和miR-138-5p表达水平;细胞计数试剂盒8（CCK-8）检测细胞增殖活性;Transwell检测细胞迁移和侵袭;蛋白质印迹（Western blot）法检测细胞周期蛋白D1（CyclinD1）、细胞周期蛋白依赖性激酶抑制剂1A（p21）、基质金属蛋白酶2（MMP-2）、基质金属蛋白酶9（MMP-9）的表达;双荧光素酶报告实验检测lncRNA LUADT1和miR-138-5p的靶向关系。结果:胶质瘤组织中lncRNA LUADT1高表达,miR-138-5p低表达。抑制lncRNA LUADT1表达或过表达miR-138-5p,细胞活性降低,细胞迁移、侵袭数降低,CyclinD1、MMP-2、MMP-9蛋白表达水平降低,p21蛋白表达水平升高。lncRNA LUADT1靶向调控miR-138-5p,干扰miR-138-5p表达逆转了抑制lncRNA LUADT1表达对胶质瘤U251细胞增殖、迁移、侵袭的抑制作用。结论:抑制lncRNA LUADT1表达通过靶向miR-138-5p抑制胶质瘤细胞的增殖、迁移和侵袭。     

lncRNAXIST/miR-34a-5p/SIRT6 分子轴调控口腔鳞癌细胞的增殖及转移

[摘要] 目的：探究lncRNA XIST/miR-34a-5p/SIRT6 分子轴调控口腔鳞癌细胞增殖和转移及其分子机制。方法：收集2013年3 月至2018 年3 月在青岛市口腔医院就诊的OSCC患者47 例癌组织和癌旁组织标本,采用qPCR检测OSCC患者组织及细胞系中lncRNA XIST、miR-34a-5p、SIRT6 mRNA 的表达,WB检测OSCC 患者组织及细胞系中SIRT6、Ki67、pcDNA、cleaved-caspase3、cleaved-caspase8、E-cadherin、Vimentin 蛋白的表达,采用CCK-8 实验检测敲降lncRNA XIST对Cal-27 及Tca-8113 细胞增殖的影响,Transwell 小室法检测Cal-27 及Tca-8113 细胞迁移及侵袭;流式细胞术检测Cal-27 及Tca-8113 细胞凋亡情况,双荧光素酶报告基因检测lncRNA XIST与miR-34a-5p、miR-34a-5p 与SIRT6 靶向结合关系。结果：lncRNA XIST和SIRT6 在OSCC患者癌组织及细胞系中高表达（均P<0.05）,miR-34a-5p 则呈低表达（P<0.01）;敲降lncRNA XIST抑制OSCC细胞的增殖、迁移及侵袭并促进细胞凋亡（均P<0.01）,同时转染miR-34a-5p 抑制剂或pcDNA-SIRT6 载体作用则相反;敲降lncRNA XIST 促进OSCC细胞中增殖及转移相关蛋白表达（均P<0.01）,同时转染miR-34a-5p 抑制剂或pcDNA-SIRT6 载体作用则相反;lncRNA XIST 与miR-34a 靶向结合,miR-34a 与SIRT6 靶向结合;lncRNA XIST 通过靶向miR-34a-5p 上调SIRT6 表达（P<0.01）。结论：lncRNA XIST/miR-34a-5p/SIRT6 分子轴能够调控OSCC细胞增殖及转移,为OSCC治疗提供潜在靶点。     

miR -450a -5p 对人卵巢癌 SKOV3细胞生物学行为的影响

目的:研究miR-450a-5p对体外培养的人浆液性卵巢癌SKOV3细胞增殖、迁移、侵袭、周期、凋亡机制的影响,分析miR-450a-5p对人卵巢癌SKOV3细胞肿瘤生物学行为的影响。方法:运用Hiperfect转染试剂介导的转染法将miR-450a-5p mimics转染人卵巢癌SKOV3细胞,设阴性对照组(NC组)及空白对照组(blank组)。采用MTT法、Transwell迁移及侵袭实验、划痕实验、流式细胞术,分别检测细胞增殖、迁移及侵袭能力、周期及凋亡。结果:MTT实验:miR-450a-5p mimics组细胞增殖活性与NC组、blank组无明显差异(P>0.05)。Transwell迁移及侵袭实验均显示转染miR-450a-5p mimics后,细胞的迁移及侵袭能力明显减弱(P<0.05)。划痕后miR-450a-5p mimics组细胞迁移能力较NC组、blank组降低(P<0.05)。流式细胞术:转染miR-450a-5p mimics后,mimics组细胞周期明显被阻滞于G1期(P<0.05)。miR-450a-5p mimics组、NC组、blank组凋亡结果无明显差异(P>0.05)。结论:过表达miR-450a-5p可能对人卵巢癌细胞系SKOV3的侵袭及迁移能力存在负性调控,可能将SKOV3细胞周期阻滞于G1期。     

lncRNA BLACAT1靶向结合miR-29a-3p调控甲状腺癌细胞TPC-1的生物学行为

目的:探讨长链非编码RNA BLACAT1(lncRNA BLACAT1)调控microRNA-29a-3p（miR-29a-3p）对甲状腺癌细胞恶性生物学行为的作用及其可能机制。方法:收集2018年06月至2019年03月在我院行甲状腺癌切除术的31例患者的肿瘤组织和相应的癌旁组织。采用qRT-PCR检测lncRNA BLACAT1、miR-29a-3p在甲状腺癌组织、癌旁组织、5种甲状腺癌细胞系（SW579、 PDTC-1、 HMGA1、 TPC-1、 KAT-5）及正常甲状腺细胞Nthy-ori 3-1中的表达水平;调控TPC-1甲状腺癌细胞系中lncRNA BLACAT1、miR-29a-3p的表达,采用CCK-8法检测细胞增殖情况,Transwell法检测细胞的迁移和侵袭能力。用生物信息分析和双荧光素酶报告基因法预测和验证lncRNA BLACAT1与miR-29a-3p的靶向关系。结果:与癌旁组织和正常甲状腺细胞相比,甲状腺癌组织和细胞系中lncRNA BLACAT1的表达水平显著上调,miR-29a-3p的表达水平显著下调（P＜0.05）;过表达lncRNA BLACAT1促进TPC-1细胞的增殖、迁移和侵袭;在甲状腺癌组织中lncRNA BLACAT1的表达水平与miR-29a-3p的表达水平呈负相关（r2 =0.492,P＜0.001）;双荧光素酶分析证实lncRNA BLACAT1能特异性结合miR-29a-3p,并能降低其表达;过表达miR-29a-3p抑制TPC-1细胞的增殖、迁移和侵袭,且miR-29a-3p可以抑制由lncRNA BLACAT1过表达引起的TPC-1细胞增殖、迁移和侵袭能力的增强。结论:lncRNA BLACAT1通过靶向调控miR-29a-3p表达影响甲状腺癌细胞的增殖、迁移和侵袭,从而促进甲状腺癌的发展。     

lncRNASNHG11通过吸附miR-193a-5p促进NSCLC细胞A549的恶性生物学行为

目的： 探讨lncRNA SNHG11对非小细胞肺癌（NSCLC）A549细胞增殖、侵袭和迁移的影响及其可能机制。 方法:qPCR检测人胚肺细胞HEL-1和NSCLC细胞A549、H1299、HCC827中lncRNASNHG11和miR-193a-5p的表达水平,向A549细胞中转染SNHG11小干扰RNA（si-SNHG11）、miR-193a模拟物（miR-193a mimic）或miR-193a抑制剂（miR-193a inhibitor）后,CCK-8法检测其对细胞增殖的影响,Transwell小室和细胞划痕实验检测对细胞侵袭和迁移的影响,WB法检测对细胞增殖抗原Ki67、细胞周期蛋白D1 （cyclin D1）表达的影响,双荧光素酶报告实验验证lncRNASNHG11与miR-193a-5p的靶向关系。 结果： 与人胚肺细胞HEL-1相比,NSCLC细胞A549、H1299、HCC827中lncRNA SNHG11均呈高表达、miR-193a-5p呈低表达（均P<0.05）;沉默lncRNA SNHG11可抑制A549细胞的增殖、侵袭和迁移,降低细胞中Ki67和Cyclin D1蛋白的表达水平（均P<0.05）;过表达miR-193a-5p可抑制A549细胞增殖和侵袭迁移（均P<0.05）。lncRNASNHG11可靶向吸附miR-193a-5p,抑制miR-139a-5p可部分逆转沉默lncRNA SNHG11对A549细胞增殖、侵袭和迁移的作用（均P<0.05）。 结论：lncRNA SNHG11通过吸附miR-193a-5p促进NSCLCA549细胞的增殖、侵袭和迁移。     

利多卡因通过调控lncRNA TTN-AS1/miR-524-5p表达对骨肉瘤细胞增殖、迁移和侵袭的影响

目的:研究利多卡因对骨肉瘤细胞MG-63增殖、迁移和侵袭的影响,并探索其作用机制。方法:使用1 mmol/L、5 mmol/L、10 mmol/L浓度利多卡因处理MG-63细胞,MTT法检测细胞增殖,Transwell小室法检测细胞迁移和侵袭,Western blot检测细胞周期蛋白D1（CyclinD1）、p21、基质金属蛋白酶-2（MMP-2）、基质金属蛋白酶-9（MMP-9）蛋白表达,qRT-PCR检测长链非编码RNA TTN-AS1（TTN-AS1）和微小RNA-524-5p（miR-524-5p）表达,starBase软件结合双荧光素酶报告实验分析TTN-AS1与miR-524-5p的靶向关系。MG-63细胞中转染si-TTN-AS1、miR-524-5p或pcDNA-TTN-AS1（并进行10 mmol/L利多卡因处理）,观察细胞的增殖、迁移、侵袭。结果:不同浓度利多卡因明显提高细胞增殖抑制率和p21蛋白表达量（P＜0.05）,显著减少迁移细胞数、侵袭细胞数和CyclinD1、MMP-2、MMP-9蛋白表达量（P＜0.05）,均呈剂量依赖性。TTN-AS1可靶向调控miR-524-5p表达。抑制TTN-AS1表达与miR-524-5p过表达显著增加MG-63细胞的增殖抑制率和p21蛋白表达量（P＜0.05）,明显降低迁移细胞数、侵袭细胞数和CyclinD1、MMP-2、MMP-9蛋白表达量（P＜0.05）。TTN-AS1过表达逆转了利多卡因对MG-63细胞增殖、迁移、侵袭和CyclinD1、MMP-2、MMP-9蛋白表达的抑制作用,以及对miR-524-5p、p21蛋白表达的促进作用。结论:利多卡因通过调控lncRNA TTN-AS1/miR-524-5p表达抑制骨肉瘤MG-63细胞增殖、迁移和侵袭。     

沉默circZFR通过调控miR-497-5p表达抑制脊索瘤细胞的增殖、迁移及侵袭

目的:研究环状RNA（circular RNAs,circRNAs）circZFR是否通过调控微小RNA（microRNA,miRNA/miR）-497-5p的表达影响脊索瘤细胞增殖、迁移及侵袭。方法:实时荧光定量PCR（quantitative real-time polymerase chain reaction,qRT-PCR）检测脊索瘤组织中circZFR和miR-497-5p的表达。在脊索瘤U-CH1细胞中转染si-circZFR或miR-497-5p。噻唑蓝（methyl thiazolyl tetrazolium,MTT）检测细胞增殖,Transwell小室法检测细胞迁移及侵袭情况,蛋白质印迹法（Western blot）检测细胞周期蛋白D1（CyclinD1）、p21、基质金属蛋白酶-2（matrix metalloprotease-2,MMP-2）、基质金属蛋白酶-9（matrix metalloprotease-9,MMP-9）蛋白表达,StarBase预测工具和双荧光素酶报告实验分析circZFR与miR-497-5p的靶向结合。si-circZFR和anti-miR-497-5p共转染,观察下调miR-497-5p表达对沉默circZFR表达诱导的U-CH1细胞增殖、迁移及侵袭的作用。结果:与髓核组织比较,脊索瘤组织中的circZFR表达量明显增加,miR-497-5p表达量显著减少（P＜0.05）。沉默circZFR表达或过表达miR-497-5p明显降低U-CH1细胞48 h、72 h的细胞活性、迁移细胞数、侵袭细胞数、CyclinD1、MMP-2、MMP-9蛋白表达量,显著提高p21蛋白水平（P＜0.05）。circZFR靶向调控miR-497-5p的表达。下调miR-497-5p表达逆转了沉默circZFR表达对脊索瘤U-CH1细胞增殖、迁移、侵袭、CyclinD1、MMP-2及MMP-9蛋白表达的抑制作用,和对p21蛋白表达的促进作用。结论:沉默circZFR表达可以抑制脊索瘤细胞的增殖、迁移及侵袭,其作用机制与靶向调控miR-497-5p的表达有关。     

地锦草乙醇提取物通过circRHOT1/miR-29a-3p轴抑制结直肠癌SW480 细胞的恶性生物学行为

目的：探讨地锦草乙醇提取物（EEEH）对人结直肠癌SW480细胞生物学行为的影响及其分子机制。方法：体外培养SW480 细胞,实验分为 Con 组、EEEH-L 组、EEEH-M 组、EEEH-H 组、si-NC 组、si-circRHOT1 组、EEEH-H+pcDNA 组、EEEH-H+pcDNA-circRHOT1组,分别以si-NC、si-circRHOT1、pcDNA、pcDNA-circRHOT1转染SW480细胞,采用CCK-8法、细胞克隆形成实验、Transwell实验分别检测转染后各组细胞的增殖、迁移及侵袭能力,qPCR 法检测转染后各组SW480细胞circRHOT1和miR-29a-3p的表达,WB法检测各组细胞中MMP-2、MMP-9蛋白的表达。双荧光素酶报告基因实验检测circRHOT1与miR-29a-3p之间的靶向关系。结果：与Con组比较,EEEH-L组、EEEH-M组、EEEH-H组SW480细胞中MMP-2和MMP-9蛋白表达均明显降低（均P<0.05）,circRHOT1的表达均降低（均P<0.05）而miR-29a-3p的表达均升高（均P<0.05）且呈剂量依赖性;细胞的存活率、细胞克隆形成数、迁移及侵袭细胞数均减少（均P<0.05）。circRHOT1可靶向负调控miR-29a-3p的表达。敲减circRHOT1可抑制SW480细胞的增殖、迁移及侵袭能力 ,而过表达 circRHOT1 则可减弱 EEEH对SW480细胞增殖、迁移及侵袭的抑制作用。结论：EEEH可通过调控circRHOT1/miR-29a-3p轴而抑制结直肠癌SW480细胞的增殖、迁移及侵袭能力。     

14-3-3 sigma possibly plays a constitutive role in papillary carcinoma, but not in follicular tumor of the thyroid

14-3-3 σ is a negative regulator of the cell cycle and contributes to G2 arrest. Thus far, the lack of its expression due to hypermethylation of the CpG islands has been reported in some carcinomas. In this study, we investigated the expression of 14-3-3 σ in thyroid neoplasms by means of immunohistochemistry as well as Western blot analysis. Normal follicules did not express 14-3-3 σ. In 82 papillary carcinomas, all the cases expressed 14-3-3 σ and its expression was not reduced but even enhanced in the advanced stage and in poorly differentiated types. Furthermore, 21 of the 23 anaplastic carcinomas expressed 14-3-3 σ and its expression level tended to be higher than in papillary carcinoma. On the other hand, none of the 34 follicular carcinomas or 29 follicular adenomas expressed 14-3-3 σ. These results suggest that 14-3-3 σ plays a constitutive role in papillary carcinoma rather than acting as a cell cycle regulator, whereas it is not required for the occurrence and development of follicular tumor.     

Regulation of 14-3-3σ expression in human thyroid carcinoma is epigenetically regulated by aberrant cytosine methylation

Increased 14-3-3sigma expression has been observed by immunohistochemistry in papillary and anaplastic tumors, but not follicular thyroid cancers. 14-3-3sigma mRNA expression and methylation status was examined in tumor cell lines and primary thyroid tissues using real-time RT-PCR, bisulfite sequencing and methylation-specific PCR. Most of the 27 CpG's in the gene's CpG island were methylated in normal thyroid, TPC-1, NPA, FTC-238 and 2-7, which did not express 14-3-3sigma. In contrast, they were unmethylated in KAK-1 and anaplastic lines KAT4 and DRO-90. 14-3-3sigma expression was not increased in thyroid carcinomas, the majority of which had a methylated CpG island. In addition, 5-aza-dC treatment increased 14-3-3sigma expression in the FTC-238 and NPA cell lines, which had low baseline expression. We conclude 14-3-3sigma expression in thyroid carcinomas is regulated by CpG island hypermethylation.     

MRP3／ABCC3的特点及其在肿瘤细胞耐药性方面的作用

化疗是肿瘤综合治疗的主要方法之一，但癌细胞的多药耐药（multidrug resistance，MDR）严重影响患者的化疗效果，耐药癌细胞的存在也是患者术后复发、远处转移直至死亡的根本原因。国内外已经发现了一些MDR相关分子，如P-糖蛋白、多药耐药相关蛋白、谷胱甘肽S转移酶、拓扑异构酶、凋亡相关蛋白等。其中多药耐药相关蛋白3（MRP3／ABCC3）显示了在多种肿瘤细胞包括肺癌中的非小细胞肺癌（non—small cell lung cancer，NSCLC）中与耐药的相关性。     

G3BP和VEGFR-3在大肠癌中的表达及意义

[目的]探讨G3BP和VEGFR-3在大肠癌中的表达及其意义。[方法]选取60例经手术切除癌组织后的大肠癌病人分别选取癌组织和癌旁组织,应用Westernblot方法分别测定G3BP和VEGFR-3在癌组织和癌旁组织中的表达,分析其相关性。[结果]大肠癌组织中G3BP表达的阳性率为70.0%,癌旁组织中未检出G3BP表达;G3BP表达与大肠癌Dukes分期(P<0.05)、淋巴结转移(P<0.05)有显著性相关。大肠癌组织VEGFR-3阳性表达率为58.3%,明显高于癌旁组织VEGFR-3的阳性表达率18.2%(P<0.05);VEGFR-3表达与大肠癌患者的Dukes分期(P<0.05)、淋巴结转移(P<0.05)有显著相关性。G3BP和VEGFR-3在大肠癌组织中表达呈正相关(r=0.376,P<0.05)。[结论]大肠癌中G3BP和VEGFR-3存在明显的高表达,与患者Dukes分期、淋巴结转移关系密切,G3BP和VEGFR-3之间存在显著性相关。     

Interferon-gamma regulation of TNFalpha-induced matrix metalloproteinase 3 expression and migration of human glioma T98G cells

Induction of proinflammatory cytokines in response to malignant cells is an integral component of immune response to control tumor development. However, recent evidences have suggested that tumor cells may evade the immune system and exploit inflammatory responses to enhance its own growth. An exemplary example is the highly invasive and tumor necrosis factor (TNF)alpha-resistant glioblastoma, whose growth is associated with TNFalpha expression. We thus examined whether the tumor takes advantage of TNFalpha overexpression to enhance its invasiveness. To delineate the contribution of inflammation in tumor migration, we demonstrated that the role of proinflammatory cytokines on matrix metalloproteinases-3 (MMP-3) expression, and its consequent effects on the invasiveness of a human glioma cell-line, T98G. By using Matrigel Invasion Chamber, T98G cell migration was significantly enhanced in response to TNFalpha. In contrast, interferon-gamma (IFN gamma) reduced both basal and TNFalpha-enhanced cell invasion. To investigate the mechanisms involved, we demonstrated that TNFalpha upregulated mRNA and protein expression of MMP-3 in T98G cells, whereas IFN gamma downregulated the MMP-3 expression. The role of MMP-3 in glioma invasiveness was further confirmed by transfecting MMP-3 siRNA in T98G to abrogate the TNFalpha-enhanced cell invasion. To delineate the mechanisms further, we showed that IFN gamma exerts an inhibitory effect on the binding of TNFalpha-activated Ets-1 and NF kappa B to their respective enhancer elements found in MMP-3 promoter. In summary, our results indicated that TNFalpha enhances the invasiveness of T98G glioma cells through MMP-3 induction, and such enhancement of cell migration can be inhibited by IFN gamma.     

Predictive value of p53 and 14-3-3sigma for the effect of chemoradiation therapy on esophageal squamous cell carcinoma

BACKGROUND AND OBJECTIVES: The p53 family regulates cell-cycle arrest, triggers apoptosis, repairs DNA damage caused by various genotoxic stresses, and protects cells from death upon irradiation. The purpose of the present study was to examine the expressions of p53 and one of the p53 family proteins, 14-3-3sigma, in biopsy specimens and to predict the clinical and histological responses to chemoradiation therapy (CRT) in patients with esophageal squamous cell carcinoma (ESCC). METHODS: We investigated with the relationship between p53 and 14-3-3sigma expressions in biopsy specimens obtained from 62 patients with ESCC and analyzed these patients' clinical and histological responses to CRT. Chemoradiation therapy consisted of 5-fluorouracil plus cisplatin and 40 Gy of radiation. RESULTS: Following CRT, 71.0% of patients showed a positive clinical response and 52.8% showed a positive histological response. The rate of positive expression was 43.5% for p53 and 58.1% for 14-3-3sigma. Statistically significant correlations were found between p53 expression and clinical response to CRT (P = 0.001) and histological response to CRT (P = 0.041), and between 14-3-3sigma expression and histological response to CRT (P = 0.01). Furthermore, in p53-positive tumors, CRT was more effective in tumors with 14-3-3sigma-positive expressions than those with 14-3-3sigma-negative expressions (P = 0.037). The survival rate of the patients with 14-3-3sigma-positive tumors was better than those with 14-3-3sigma-negative tumors in patients with p53-positive tumors (P = 0.047). CONCLUSIONS: We demonstrated that p53-negative or 14-3-3sigma-positive expressions were closely related to the response to CRT. It is clinically useful to examine the expression of these genes in biopsy specimens for predicting the CRT outcomes in patients with ESCC.     

IFN-gamma withdrawal after immunotherapy potentiates B16 melanoma invasion and metastasis by intensifying tumor integrin alphavbeta3 signaling

Immunotherapy can effectively suppress tumor, yet complete tumor eradication occurs infrequently. The metastatic potential of remnant tumor cells after immunotherapy and the underlying mechanisms have not been fully elucidated. Here, we report that the termination of immunotherapy strikingly increases the metastatic potential of remnant melanoma. This is mainly due to the withdrawal of IFN-gamma after immunotherapy. The relief of IFN-gamma stress led to the increase of alphavbeta3 integrin expression in B16 cells, which increased the adhesion of B16 cells to fibrinogen, fibronectin and laminin. Through alphavbeta3 signaling, the activation of FAK, upregulation of cdc2, production of active MMP-2 and MMP-9 and actin polymerization were intensified in B16 cells stimulated with ECM molecules 24 h after the withdrawal of IFN-gamma. The i.v. injection of such tumor cells into mice resulted in more metastatic tumor nodes in lung and shortened the survival of mice. The pitfall of immunotherapy termination can be remedied by the administration of recombinant CBD-HepII polypeptide of fibronectin, which effectively inhibits alphavbeta3 signaling. These findings suggest that the risk of tumor metastasis can be increased after the termination of immunotherapy, due to the withdrawal of IFN-gamma and that targeting alphavbeta3 signaling pathway can improve the therapeutic effect of immunotherapeutic approaches by reducing such metastatic risk.     

人肝癌细胞感染蓝舌病毒HbC_3株超微结构的动态观察

目的探讨蓝舌病毒靶向抗肿瘤的细胞生物学机制。方法利用透射电镜观察蓝舌病毒HbC3株感染人肝癌细胞Hep-3B的形态发生学以及该病毒引起的细胞的病理改变。结果BTV-HbC3以受体介导的胞饮作用穿入细胞,溶酶体水解病毒外衣壳,使之成为亚病毒粒子,胞浆内有病毒包涵体及未装配成熟的亚病毒颗粒。随后亚病毒颗粒装配上外层蛋白结构,形成成熟的病毒粒子。病毒感染细胞12~18h时,细胞以挤出的方式释放病毒并达到高峰。18~48h时,病毒进入超感染期,大量细胞发生病变,出现细胞凋亡和溶解。结论一旦蓝舌病毒感染Hep-3B肿瘤细胞即可在细胞内增殖并诱导该肿瘤细胞进入凋亡,死亡的肿瘤细胞释放出的病毒粒子并再次感染其他肿瘤细胞,直至将全部肿瘤细胞杀灭,其溶瘤方式为链式反应。     

腹腔热化疗中卵巢癌细胞对顺铂敏感性影响的体外研究

目的:探讨腹腔热疗对卵巢癌细胞顺铂敏感株的化疗增敏作用及对顺铂耐药株的逆转耐药作用,并分析其作用机制,为腹腔热疗增加顺铂化疗敏感性及逆转卵巢癌顺铂耐药提供实验依据。方法:MTT 法研究常温及41℃不同作用时间下温热联合顺铂对卵巢癌细胞敏感株SKOV 3、卵巢癌细胞顺铂耐药株SKOV 3/DDP 的生长抑制作用。Western bloting法检测常温及41℃不同作用时间下ERCC1 基因表达水平。结果:41℃下作用60、90min时对SKOV 3 细胞生长的抑制作用大,差异有显著性,P<0.001。热处理对SKOV 3/DDP 细胞生长有抑制作用,不同时间差异有统计学意义。热疗联合顺铂在常温或41℃下,SKOV 3 细胞对顺铂的敏感性不同,差异有统计学意义（P<0.001）。 且41℃作用90min时敏感性最大（P=0.002）,对顺铂的敏感性提高79.885% 。热处理不同时间,SKOV 3/DDP 细胞对顺铂的敏感性增加,差异有显著性,P<0.001。41℃90min SKOV3/DDP 细胞对顺铂的敏感性提高37.129% 。在SKOV 3 细胞中,ERCC1 表达水平较SKOV 3/DDP 细胞降低,F=32.175,P<0.001。热疗联合顺铂与常温下相比,SKOV 3、SKOV 3/DDP 细胞中ERCC1 基因表达水平差异无统计学意义,F=0.962,P=0.443。结论:热疗对卵巢癌细胞SKOV 3 及其顺铂耐药亚株SKOV 3/DDP 细胞有生长抑制作用;热疗联合顺铂可以增加SKOV 3 细胞对顺铂的敏感性,可以部分逆转SKOV 3/DDP细胞对顺铂的耐药性,且最佳作用时间为41℃90min;ERCC1 表达增加与卵巢癌顺铂耐药有关,但热疗联合顺铂对ERCC1 基因表达水平无明显影响,热疗可能通过其他途径增加肿瘤细胞对顺铂的敏感性。