miR-183调控Wnt/β-catenin信号通路影响胃癌SGC-7901细胞生物学特性的研究

背景 miR-183在胃癌、乳腺癌、膀胱癌等多种肿瘤组织中低表达,发挥抑癌基因的作用,但其在胃癌中作用机制目前尚不十分清楚.研究表明, miR-183可以通过调节Wnt/β-catenin信号通路抑制骨肉瘤细胞的生长、迁移和侵袭,而Wnt/β-catenin信号通路在胃癌中高度激活与胃癌的发生和转移密切相关.但miR-183是否调节Wnt/β-catenin信号通路影响胃癌细胞生物学特性尚不清楚.目的探讨miR-183调控Wnt/β-catenin信号通路对胃癌细胞生物学特性的影响.方法采用q RT-PCR检测miR-183在不同胃癌细胞株中的表达情况,在胃癌细胞SGC-7901中转染miR-183mimics或mimics对照,分别设为miR-183组和miR-NC组, qRT-PCR检测转染效率,噻唑蓝增殖实验检测SGC-7901细胞增殖变化,流式细胞仪检测SGC-7901细胞凋亡情况, Transwell实验检测SGC-7901细胞侵袭和迁移能力, Western blot法检测凋亡相关及Wnt/β-catenin信号通路相关蛋白表达水平.使用Wnt/β-catenin信号通路激动剂氯化锂处理过表达miR-183的SGC-7901细胞,观察SGC-7901细胞生物学特性的变化.结果与正常胃黏膜上皮GES-1细胞相比, 4株胃癌细胞中miR-183的表达水平明显降低(P 0.05).转染miR-183mimics后SGC-7901细胞中miR-183的表达水平显著升高(P0.05).过表达miR-183后SGC-7901细胞OD值降低(P0.05),凋亡率、Bax和Cleaved Caspase-3蛋白表达水平升高(P 0.05),侵袭和迁移细胞数减少(P0.05),β-catenin、p-GSK-3β和Cyclin D1蛋白表达水平下调(P0.05), GSK-3β蛋白表达水平上调(P 0.05).激活Wnt/β-catenin信号通路部分逆转了过表达miR-183对SGC-7901细胞增殖、侵袭和迁移的抑制作用及凋亡促进作用(P0.05).结论miR-183可能通过抑制Wnt/β-catenin信号通路阻碍人胃癌SGC-7901细胞增殖、侵袭和迁移能力,促进细胞凋亡.     

奥曲肽对SGC-7901胃癌细胞中Fas,FasL及P53表达的影响

目的:探讨奥曲肽(OCT)对胃癌细胞SGC-7901中Fas,FasL,P53,P21,P27和C-Myc表达的影响.方法:流式细胞术和RT-PCR法分别用于检测经OCT(1×10-7 mol/L)处理前后胃癌细胞SGC-7901中Fas,FasL,P53,P21,P27,c-Myc阳性癌百分率、mRNA表达和蛋白表达及细胞周期G0/G1、S、G2/M的变化.结果:胃癌细胞SGC-7901经OCT诱导6、12、24和48 h垢,Fas和FasL mRNA表达均明显增加,P53 mRNA表达则显著减少,而P21,P27和c-Myc mRNA表达均无明显变化;胃癌细胞SGC-7901经OCT诱导12 h后,Fas和FasL蛋白相对表达强度也显著增加,P53蛋白相对表达强度则明显降低(5.5±0.3 vs 3.2±0.1,5.1±0.3 vs 4.5±0.1.3.3±0.2 vs 4.9±0.3,P＜0.05或0.01),而P21,P27和c-Myc蛋白相对表达强度均无明显改变.细胞周期G0/G1、S、G2/M在OCT处理前后的变化并不明显.结论:OCT能上调SGC-7901胃癌细胞中Fas和FasL表达,下调突变型p53表达,是其诱导凋亡作用的重要机制之一.     

抗Fas mAb和IFN联合诱导胃癌细胞凋亡的实验研究

目的观察联合应用抗Fas单克隆抗体(mAb)和干扰素-γ(IFNγ)诱导人胃癌细胞系SGC-7901细胞凋亡,并探讨其在胃癌治疗中的意义.方法应用细胞形态观察、琼脂糖凝胶电泳、流式细胞光度术检测抗Fas mAb,IFN-γ单独及联合应用诱导的人胃癌细胞系SGC-7901细胞凋亡,并应用流式细胞光度术检测IFN-γ对人胃癌细胞系SGC-7901细胞表达Fas抗原的影响.结果抗Fas mAb显著诱导SGC-7901细胞凋亡(19.3%),细胞DNA裂解片段呈现典型的"阶梯状"排列的条带.联合应用抗Fas mAb和IFN-γ处理人胃癌细胞系SGC-7901细胞凋亡率(29.4%)显著高于对照组、IFN-γ及抗Fas mAb处理组(1.2%,1.9%,19.3%,t=17.345,17.276,5.425,P＜0.01及P＜0.05).IFN-γ处理组人胃癌细胞系SGC-7901细胞Fas抗原的表达阳性细胞数(59.3%)显著高于对照组(27.1%,t=12.995,P＜0.05),抗Fas mAb诱导SGG7901细胞凋亡的敏感性与Fas抗原的表达水平显著相关.结论抗Fas mAb可以诱导SGC-7901细胞凋亡.联合应用抗FasmAb和IFN-γ具有协同诱导人胃癌细胞凋亡的作用,其机制可能与干扰素-γ显著上调人胃癌细胞系SGC-7901细胞Fas抗原的表达水平有关.     

下调miR-221对胃癌细胞SGC-7901增殖的影响及其相关机制研究

目的探讨下调miR-221后对胃癌细胞SGC-7901增殖的影响及其相关机制。方法收集2015年6月至2017年7月我院就诊的48例胃癌患者的癌旁组织、癌组织和32例同期体检健康者的正常胃组织,采用Real-time Q-PCR检测miR-221的表达水平。体外培养胃癌细胞SGC-7901,根据处理的方法不同分为组1(mimics对照组)、组2(hsa-miR-221组)、组3(antimiR组)、组4(anti-miR-221组),运用Real-time Q-PCR检测四组miR-221的表达,并通过噻唑蓝(MTT)比色法检测细胞的增殖,同时运用免疫印迹法(Western blotting)检测细胞增殖的相关信号通路STAT3蛋白的改变。结果胃癌组织miR-221阳性表达率显著高于癌旁组织、正常组织(均P 0. 05),癌旁组织miR-221阳性表达率高于正常组织,但差异无统计学意义(P 0. 05)。胃癌SGC-7901细胞中的miR-221表达水平高于正常胃上皮细胞(4. 56±1. 21 vs. 15. 32±2. 74),差异有统计学意义(P 0. 01)。组2的增殖能力最强,组4的增殖能力最弱,组1和组3两组无明显差异(P 0. 05)。四组细胞中STAT3蛋白的相对表达量分别为0. 341±0. 028、0. 748±0. 052、0. 376±0. 031、0. 218±0. 022,组2的STAT3蛋白表达水平最高,组4 STAT3表达水平最低,组1和组3两组无明显差异(P 0. 05)。结论胃癌组织和胃癌细胞中miR-221的表达明显升高,下调miR-221可抑制胃癌细胞SGC-7901的增殖,其机制可能与下调STAT3蛋白有关。     

GRP78在胃癌中表达水平及沉默表达后对人胃癌SGC-7901/DDP细胞增殖和凋亡的影响

目的研究内质网分子伴侣葡萄糖调节蛋白78(glucose regulated protein 78,GRP78)在人不同胃组织(正常胃组织和胃癌组织)和细胞(正常胃上皮GES1细胞、胃癌SGC-7901细胞、多耐药性胃癌SGC-7901/DDP细胞)中的表达水平;揭示GRP78沉默表达对SGC-7901/DDP细胞增殖和凋亡的影响。方法利用实时荧光定量PCR(Real-time PCR)和免疫印迹(Western blotting)方法分别检测人正常胃组织、胃癌组织、人正常胃上皮GES1细胞、胃癌SGC-7901细胞、多耐药性胃癌SGC-7901/DDP细胞中GRP78 mRNA和蛋白表达水平。利用LipfectamineTM2000将GRP78 siRNA转染至SGC-7901/DDP细胞中,同时设置未转染Control组和无义转染Control siRNA组。Real-time PCR和Western blotting方法检测各组SGC-7901/DDP细胞转染效率;MTT方法检测各组SGC-7901细胞的生长活力变化和不同浓度顺铂DDP作用下细胞增殖抑制率;流式细胞术检测各组SGC-7901细胞凋亡情况。结果胃癌组织中GRP78 mRNA和蛋白表达水平显著高于正常胃组织(P0.05);多耐药性胃癌SGC-7901/DDP细胞中GRP78 mRNA和蛋白表达水平显著高于胃癌SGC-7901细胞和正常胃上皮GES1细胞(P0.05);GRP78 siRNA转染SGC-7901/DDP细胞后GRP78 mRNA和蛋白表达水平显著下降(P0.05);与Control组和Control siRNA组相比,GRP78 siRNA转染组SGC-7901/DDP细胞生长活力显著下降,DDP作用下细胞增殖抑制率和细胞凋亡率显著增加(P0.05)。结论 GRP78 mRNA和蛋白在胃癌组织中高表达,在多耐药性胃癌SGC-7901/DDP细胞中也高表达,证实GRP78在胃癌中发挥促癌基因的作用。而GRP78 siRNA转染能够沉默SGC-7901/DDP细胞中GRP78基因表达,抑制SGC-7901/DDP细胞增殖并促进细胞凋亡。     

下调miR-25抑制胃癌细胞增殖和诱导其凋亡的作用研究

目的探讨下调miR-25对胃癌SGC-7901细胞增殖凋亡的影响及可能的作用机制。方法将SGC-7901细胞分为对照组(不做处理)、阴性组(转染miR-25 inhibitor control)和干扰组(转染miR-25 inhibitor)3组,利用实时荧光定量PCR(qRT-PCR)检测转染效果,采用四甲基偶氮唑盐比色法(MTT法)检测miR-25对SGC-7901细胞增殖影响,流式细胞仪检测miR-25对细胞凋亡的影响,蛋白质免疫印迹法(Western blotting)检测各组细胞中PTEN蛋白表达量。结果转染miR-25 inhibitor后,细胞中miR-25的表达量较对照组显著降低(P0.05),阴性组和对照组间差异无统计学意义(P0.05)。与对照组相比,干扰组中细胞存活率显著降低,凋亡率显著升高,差异均有统计学意义(P0.05)。干扰组细胞中PTEN蛋白的表达量较对照组明显增加(P0.05),对照组和阴性组间差异无统计学意义(P0.05)。结论下调miR-25可抑制胃癌SGC-7901细胞增殖和诱导细胞凋亡,其作用机制可能与上调PTEN蛋白的表达有关。     

木黄酮对人胃癌细胞SGC-7901作用的研究

目的探讨木黄酮对体外培养的人胃癌细胞SGC-7901生长的抑制和诱导细胞凋亡的作用。方法用MTT法检测药物效应，透射电镜及流式细胞仪观察木黄酮处理SGC-7901细胞后细胞周期和细胞凋亡。细胞免疫化学观察木黄酮处理SGC-7901细胞后，SGC-7901细胞PCNA，FAS，C-mvc的变化。结果①MTT法证实木黄酮对SGC-7901细胞生长有抑制作用，并呈剂量-效应关系。②流式细胞仪分析木黄酮处理SGC-7901细胞后细胞滞留于G2／M期，并可见凋亡峰。③透射电镜可见SGC-7901细胞出现典型的细胞凋亡及坏死形态学改变。④木黄酮下调PCNA及C—myc表达，上调FAS表达。结论木黄酮对人胃癌细胞SGC-7901有抑制作用，其抑制作用可能通过下调C-mvc基因表达，上调Fas蛋白表达，下调PCNA表达，从而多途径诱导胃癌细胞凋亡和坏死、阻抑细胞周期进程，抑制SGC-7901细胞增殖。     

西咪替丁对胃癌细胞SGC-7901增殖及凋亡的影响

目的:观察西咪替丁对人胃癌SGC-7901细胞增殖、细胞周期分布及凋亡的影响,并初步探讨其作用机制.方法:培养人胃癌SGC-7901细胞,以不同浓度的西咪替丁处理后用MTT法检测SGC-7901细胞的增殖情况;流式细胞术检测癌细胞周期和凋亡;Hoechst33258染色后荧光显微镜观察药物作用后癌细胞的形态变化;透射电镜观察用药后细胞超微结构的改变;Western印记法检测Bcl-2和Bax蛋白表达.结果:以不同浓度的西咪替丁分别处理人胃癌SGC-7901细胞24h和48 h,结果发现,在0.5,1,2.5,5,10 mmol/L时对SGC-7901细胞的增殖具有显著的抑制作用,与对照组相比差异显著(24 h:0.705±0.018,0.560±0.038,0.408±0.029,0.276±0.042,0.205±0.031 vs 0.803±0.012,P<0.05;48 h:0.902±0.024,0.671±0.015,0.420±0.030,0.180±0.037,0.0117±0.021 vs 1.079±0.040,P<0.05),并呈时间和剂量依懒性,而在0.25 mmol/L以下浓度对SGC-7901细胞未见明显细胞毒作用;0.5-10 mmol/L西咪替丁作用后,可观察到典型的细胞凋亡形态学改变;流式细胞仪检测可见凋亡峰,G0/G1期细胞明显增多(60.83±2.27,67.21±1.18,75.15±4.01,81.88±3.10,86.99±1.43 vs 50.28±1.97,P<0.05);西咪替丁还可下调SGC-7901细胞中的Bcl-2蛋白表达,上调Bax蛋白表达.结论:西咪替丁可改变细胞周期分布,并能通过下调Bcl-2、上调Bax蛋白表达,诱导SGC-7901细胞凋亡,从而抑制细胞增殖.     

MicroRNA-129通过靶向调控HMGA2抑制胃癌细胞增殖、侵袭

背景:越来越多的研究表明microRNA在胃癌发生、发展中起重要作用。有研究表明miR-129在胃癌组织中表达异常,但其对胃癌细胞增殖、侵袭的作用仍不明确。目的:探讨miR-129在胃癌组织和胃癌细胞株中的表达及其影响胃癌细胞增殖和侵袭的机制。方法:收集82例胃癌组织及其相应癌旁组织,培养人胃黏膜上皮细胞株和不同的胃癌细胞株,以q PCR法检测miR-129表达。将miR-129 mimic或miR-NC转染胃癌SGC-7901细胞后,转染HMGA2过表达质粒。以克隆形成实验观察细胞增殖情况,Transwell小室法检测细胞侵袭情况,Pearson相关分析评估miR-129表达与HMGA2表达的相关性,荧光素酶实验检测荧光素酶活性,q PCR和蛋白质印迹法分别检测miR-129、HMGA2 mRNA和蛋白表达。结果:与癌旁组织相比,胃癌组织中miR-129表达明显下降(P0.05);与人胃黏膜上皮细胞株GES-1相比,各胃癌细胞株中miR-129表达明显下降(P0.05)。与miR-NC组相比,miR-129 mimic组SGC-7901细胞增殖、侵袭能力均明显下降(P0.05)。胃癌组织中HMGA2 mRNA表达明显增加(P0.05),并与miR-129表达呈负相关(r=-0.543 9,P0.01)。野生型miR-129 mimic组荧光素酶活性显著低于miR-NC组(P0.05);转染miR-129 mimic后HMGA2 mRNA和蛋白表达显著降低(P0.05)。与miR-129+阴性对照组相比,miR-129 mimic+HMGA2组细胞增殖、侵袭能力明显升高(P0.05)。结论:miR-129在胃癌组织和细胞中低表达,其可通过下调HMGA2抑制SGC-7901细胞的增殖和侵袭。     

HSP90抑制剂17-DMAG对胃癌细胞株SGC-7901增殖及凋亡的影响

目的:探讨热休克蛋白90(HSP90)功能特异性抑制剂17-DMAG对胃癌细胞SGC-7901增殖及凋亡的影响.方法:体外培养人胃癌细胞SGC-7901,以不同浓度的17-DMAG处理后采用MTT法检测SGC-7901细胞的生长抑制情况,流式细胞仪碘化丙啶(PI)染色分析细胞周期分布,流式细胞仪及Annexin V-FITC试剂盒监测17-DMAG对胃癌细胞SGC-7901凋亡的影响.结果:不同浓度的17-DMAG对人胃癌细胞SGC-7901有明显的生长抑制作用,各组之间比较(P＜0.01),且呈时效量效依赖关系(F=241.313,246.856,均P＜0.001).17-DMAG作用24 h后胃癌细胞株SGC-7901呈现G2/M期阻滞,试验组与对照组比较(F=231.991,P＜0.001),呈浓度依赖关系.17-DMAG处理胃癌细胞SGC-7901 24 h早期凋亡细胞增加,48 h晚期凋亡细胞增加.结论:17-DMAG可明显抑制胃癌细胞SGC-7901的增殖,使胃癌细胞SGC-7901阻滞于G2/M期,诱导胃癌细胞SGC-7901凋亡.     

miR-29对胃腺癌细胞侵袭力的影响

目的探讨miR-29对胃腺癌细胞侵袭力的影响,以期为胃癌的转移、侵袭机制提供线索。方法采用实时荧光定量PCR技术检测46例患者胃腺癌标本及癌旁组织中miR-29表达,分析与患者临床病理特征的关系。结果 PCR反应检测结果显示miR-29a、miR-29b、miR-29c在胃癌组织中相对含量分别为14.13±5.78、9.81±1.95、16.77±2.04,在癌旁组织中分别为31.64±10.26、26.88±5.03、52.57±8.02,胃癌组织中的miR-29相对含量明显低于癌旁组织(P0.05);将胃癌组织标本与癌旁组织标本中的miR-29相对含量计算比值(C/P),患者不同性别、年龄、TNM分期、浸润深度、分化程度时C/P值相比,差异无统计学意义(P0.05);存在淋巴结转移时miR-29a C/P值明显低于无淋巴结转移时(P0.05),存在远处转移时miR-29b、miR-29c C/P值明显低于无远处转移时(P0.05)。结论胃腺癌组织中miR-29表达明显低于癌旁组织,miR-29a在淋巴结转移及miR-29b、miR-29c在远处转移患者胃腺癌组织中表达降低,提示其可能与胃癌的侵袭及转移有关。     

Down-regulation of miR-141 in gastric cancer and its involvement in cell growth

Purpose  Human microRNA-141 (miR-141), a member of the miR-200 family, has been reported to be associated with various human malignancies. However, it remains unknown whether miR-141 is involved in the pathogenesis of gastric cancer. Therefore, we examined the expression of miR-141 in gastric cancer tissues and the effect of miR-141 overexpression on cancer cell proliferation. Methods  The expression level of miR-141 in 35 pair-matched gastric neoplastic and adjacent non-neoplastic tissues, and in 5 gastric cancer cell lines were examined by quantitative real-time PCR. The growth of MGC-803 cells transfected with miRNA precursor was examined by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazoliumbromide) assay. Results  MiR-141 was significantly down-regulated in 80% (28/35) of primary gastric cancer tissues compared with pair-matched adjacent non-tumor tissues (P < 0.01). The expression of miR-141 was also found to be substantially reduced in several human gastric cancer cell lines such as MGC-803, HGC-27, SGC-7901 and BGC-823 cells. Overexpression of miR-141 with its precursors significantly inhibited the proliferation of gastric cancer cells. Conclusions  These results suggest that miR-141 may be involved in the development of gastric cancer through its inhibitory effect on cell proliferation. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Effect of Fibulin-5 on cell proliferation and invasion in human gastric cancer patients

Objective:To explore the effect of Fibulin-5 expression on cell proliferation and invasion in human gastric cancer patients.Methods:Fibulin-5 expression was detected in 56 samples of surgically resected gastric cancer and paired noncancerous tissues using qRT-PCR and immunoblotting.Fibulin-5 was knocked dowu by Fibulin-5 shRNA in MGC-803 cells,then Brdu cell proliferation and transwell invasion assays were used to determine cell proliferation and invasion.Results:The level of Fibulin-5 mRNA in gastric cancer tissues was significantly higher as compared with that in normal tumor-adjacent tissues(P0.05).Otherwise,the level of Fibulin-5 protein in cancer and noucancerous tissues was consistent with mRNA expression(P0.05).Fibulin-5 protein expression in tumor tissues with poorly differentiated.lymph node metastasis and advanced TNM tumor stage was significantly higher(P0.05.respectively).Fibulin-5 was obviously knocked down by Fibulin-5 shRNA(P0.05).and Fibulin-5 knockdown significantly inhibited cell proliferation and invasion in MGC-803 cells(P0.05.respectively).Conclusions:The high-expression of Fibulin-5 is associated with the malignant clinicopathologic parameters in gastric cancer and Fibuliu-5 knockdown inhibits cell proliferation and invasion in MGC-803 cells.suggesting Fibulin-5 may act as a key factor in the progression of gastric cancer.     

PTEN的表达及其在阿霉素诱导胃癌细胞凋亡中的意义

目的 研究阿霉素干预胃癌细胞后PTEN的表达及其在阿霉素诱导胃癌细胞凋亡中的意义.方法 (1)阿霉索干预胃癌细胞BGC-823后以四甲基偶氮唑盐比色法(MTT法)和流式细胞法检测细胞存活率及凋亡率,并检测VFEN的mRNA和蛋白水平.(2)构建胃癌裸鼠异位种植瘤,采用原位末端标记(TUNEL)法检测异位种植瘤中胃癌细胞的凋亡情况,并用RT-PCR和Western blot法检测PTEN mRNA和蛋白的表达水平.(3)以PTEN特异性小干扰RNA(siRNA)转染BGC-823细胞,并以阿霉素进行干预,检测BGC-823细胞的存活率和凋亡率以及PTEN蛋白表达水平.结果 (1)阿霉素干预后,BGC-823细胞的生存率呈时间依赖性降低.(2)阿霉素能够有效诱导BGC-823细胞凋亡.(3)阿霉素在BGC-823细胞中可时间依赖性地促进PTEN的mRNA和蛋白水平的升高.裸鼠异位种植瘤试验中,阿霉素干预组的凋亡率[(28.11±1.05)%]明显高于对照组[(2.78±1.63)%];阿霉素干预组瘤体组织中PTEN mRNA和蛋白水平亦高于对照组(0.5667±0.0043比0.2217±0.0063,0.14±0.26比0.04±0.15,P值均<0.05).(4)转染与未转染[WEN siRNA的胃癌细胞以阿霉素干预后,PTEN siRNA转染组的PTEN蛋白表达水平明显低于对照组(P<0.0001),且PTEN siRNA转染组[(10.35±1.04)%]凋亡率明显小于未转染组[(31.37±3.58)%],P<0.05.结论 阿霉素干预胃癌细胞后可以抑制其生长,诱导细胞凋亡,PTEN表达水平的升高可能是其机制之一.     

miR-574-3p在结直肠癌中的表达和作用机制研究

目的检测并分析结直肠癌组织和细胞系中miR-574-3p的表达情况及其对结直肠癌细胞发生、发展的影响。方法收集武汉市第三医院11对手术切除结直肠癌患者的癌组织和对应癌旁组织,3种结直肠癌细胞系和1种正常结直肠上皮细胞,通过实时荧光定量聚合酶链反应(qRT-PCR)检测临床样本及结直肠癌细胞系中miR-574-3p的表达水平;通过转染miR-574-3p mimic及相应对照mimic NC实现上调结直肠癌细胞(HCT-8、HCT-116)中miR-574-3p的表达水平,采用CCK-8、EdU、克隆形成实验、流式凋亡实验和流式细胞周期实验检测过表达miR-574-3p后对结直肠癌细胞的增殖、凋亡和细胞周期的影响。结果癌旁组织中miR-574-3p的表达差异倍数显著高于对应癌组织(P<0.05),在3种结直肠癌细胞系中miR-574-3p的表达较正常结肠上皮细胞也显著降低(P<0.05)。CCK-8、EdU、克隆形成实验表明,过表达miR-574-3p抑制结直肠癌细胞的增殖能力。流式凋亡实验结果显示,过表达miR-574-3p促进结直肠癌细胞的凋亡能力。流式细胞周期结果显示,过表达miR-574-3p使结直肠癌细胞发生G₀/G₁期阻滞。结论结直肠癌组织和细胞系中的miR-574-3p均低表达,上调结直肠癌细胞(HCT-8、HCT-116)中miR-574-3p可以抑制细胞增殖,促进凋亡并发生G₀/G₁期阻滞。     

长链脂酰辅酶A合成酶1在肝癌中的表达及临床意义

目的探讨长链脂酰辅酶A合成酶1(Long-Chain Acyl-CoA Synthetase 1,ACSL1)在肝细胞癌(hepatocellular carcinoma,HCC)的表达及临床意义。方法收集334例诊断为HCC患者的癌组织与对应癌旁组织;免疫组化分析癌组织与癌旁组织中ACSL1的表达差异,并进一步分析HCC中ACSL1表达与患者临床病理特征的关系及对患者生存的影响;慢病毒转染构建ACSL1过表达的SNU739细胞;MTS、流式细胞术检测ACSL1对SNU739细胞的增殖和凋亡的影响;SeahorseXF24分析仪、乳酸试剂盒探究ACSL1对SNU739细胞能量代谢的影响。结果HCC组织中的ACSL1表达水平显著低于癌旁组织(3.02±2.57 vs 7.60±3.34,t=20.040,P=0.000);HCC中ACSL1的表达水平与患者的年龄有相关性(c2=16.472,P=0.000);ACSL1蛋白表达水平是HCC患者生存的危险因素(HR=1.642,P=0.012);ACSL1可以显著增强SNU739细胞凋亡(t=13.886,P=0.000)、线粒体基础氧耗速率(oxygen consumption rate,OCR)(F=83.429,P=0.001)以及线粒体最大氧耗率(F=859.792,P=0.000)。结论ACSL1在HCC组织中低表达,ACSL1低表达的HCC患者预后更差;ACSL1可作为HCC患者预后的潜在标志物,有助于筛查早期HCC,提高HCC患者生存率。     

Down-expression of tumor protein p53-induced nuclear protein 1 in human gastric cancer

AIM: Overexpression of tumor protein p53-induced nuclear protein 1 (TP53INP1) induces G1 cell cycle arrest and increases p53-mediated apoptosis. To clarify the clinical importance of TP53INP1, we analyzed TP53INP1 and p53 expression in gastric cancer. METHODS: TP53INP1 and p53 expression were examined using immunohistochemistry in 142 cases of gastric cancer. The apoptosis of gastric cancer cells was analyzed using the TUNEL method. The relationship between the expression of TP53INP1 and clinicopathological factors was statistically analyzed. RESULTS: TP53INP1 was expressed in 98% (139/142 cases) of non-cancerous gastric tissues and was down-expressed in 64% (91/142 cases) of gastric cancer lesions from the same patients. TP53INP1 expression was significantly decreased (43.9%) in poorly differentiated adenocarcinoma compared with well or moderately differentiated adenocarcinoma (81.6%). Cancers invading the submucosa or deeper showed lower positively (59.1%) compared with mucosal cancers (85.2%). Decrease or loss of TP53INP1 expression was significantly correlated with lymphatic invasion (54.3% vs 82.0% without lymphatic invasion) and node-positive patients (31.3% vs 68.3% in node-negative patients). P53 was expressed in 68 (47.9%) patients of gastric cancer, whereas it was absent in normal gastric tissues. A significant association was also observed between TP53INP1 status and the level of apoptosis in tumor cells: the apoptotic index in TP53INP1-positive tissues was significantly higher than that in TP53INP1-negative portions. Finally, when survival data were analyzed, loss of TP53INP1 expression had a significant effect in predicting a poor prognosis (P=0.0006). CONCLUSION: TP53INP1-positive rate decreases with the progression of gastric cancer. TP53INP1 protein negativity is significantly associated with aggressive pathological phenotypes of gastric cancer. TP53INP1 is related to the apoptosis of gastric cancer cells. The decreased expression of the TP53INP1 protein may reflect the malignant grade of gastric cancer and is regarded as an adverse prognostic factor.     

Association of serum vascular endothelial growth factor-C and lymphatic vessel density with lymph node metastasis and prognosis of patients with gastric cancer

AIM: To investigate whether serum vascular endothelial growth factor-C (SVEGF-C), VEGF-C, and lymphatic vessel density (LVD) in tumor tissues are related to lymph node metastasis (LNM) and prognosis in gastric cancer.METHODS: SVEGF-C levels of 80 gastric cancer patients and 20 healthy donors were examined using ELISA. VEGF-C expression and LVD were examined using immunohistochemical staining. Kaplan-Meier survival analysis was performed to determine their influence on the prognosis of the patients. RESULTS: The SVEGF-C level in gastric cancer patients (595.9 ± 201.0 ng/L) was significantly higher (P = 0.000) than controls (360.0 ± 97.4 ng/L). Both SVEGF-C and LVD were significantly higher in poorly differentiated adenocarcinomas, T3 and T4, LNM, distant metastasis, and pTNM groups Ⅲ and IV (P = 0.000). The sensitivity and specificity of SVEGF-C for predicting LNM were 82.8% and 81.8%, respectively (cut-off = 542.5 ng/L). The positive expression rate of VEGF-C was significantly higher in cancerous than in normal tissues (65% vs 20%; P = 0.001). VEGF-C expression up-regulation was significantly related to differentiation, depth of invasion, LNM, distant metastasis, and pTNM stage (P = 0.000). LVD was 10.7 ± 3.1/200 HP in the experimental group vs 4.9 ± 1.3/200 HP in controls (P = 0.000); LVD in cancerous tissues with and without LNM was 12.0 ± 2.7/200 HP vs 7.6 ± 0.5/200 HP, respectively (P = 0.000). SVEGF-C and LVD were significantly higher in VEGF-C positive than in negative patients (P=0.000); SVEGF-C level was related to LVD (P = 0.000). Kaplan-Meier survival analysis factors predicating poor prognosis were: SVEGF-C level (P = 0.001), VEGF-C expression and LVD (both P = 0.000). CONCLUSION: SVEGF-C level, VEGF-C and LVD are related to LNM and poor prognosis of patients with gastric cancer. SVEGF-C may be a biomarker for LNM in gastric cancer.     

Growth,invasion, metastasis,differentiation, angiogenesis and apoptosis of gastric cancer regulated by expression of PTEN encoding products

AIM: To investigate expression of PTEN in gastric cancer and to explore its roles in tumorigenesis and progression of gastric cancer.METHODS: Formalin-fixed and paraffin-embedded tissues of adjacent non-tumor mucosa and primary foci from 113cases of gastric cancers were studied for the expression of PTEN and Caspase-3 andmicrovessel density (MVD)by streptavidin-peroxidase (S-P) immunohistochemistry with antibodies against PTEN, Caspase-3, and CD34. The relationship between PTEN and Caspase 3 expression and clinicopathological parameters of tumors was compared.RESULTS: Primary gastric cancer cells expressed PTEN less frequently than adjacent epithelial cells of primary foci (54.9％ vs89.4％; P=0.000, χ2=33.474). PTEN expression was significantly associated with invasive depth (P=0.003,rs=0.274), metastasis (P=0.036, rs=0.197), growth pattern (P=0.008, rs=0.282), Lauren′s classification (P=0.000,rs=0.345), and histological classification (P=0.005, rs=0.262)of tumors, but not with tumor size (P=0.639, rs=0.045),Borrmann′s classification (P=0.544, rs=0.070) or TNM staging (P=0.172, rs=0.129). PTEN expression was negatively correlated with MDV in primary gastric cancer (P=0.020,F=5.558). Primary gastric cancer cells showed less frequent immunoreactivity to Caspase-3 than adjacent epithelial cells of primary foci (32.7 ％ vs 50.4 ％; P=0.007,χ2=7.286).Caspase-3 expression was dependent of PTEN expression in primary gastric cancer cells (P=0.000, χ2=15.266).CONCLUSION: Down-regulated expression of PTEN plays an important role in tumorigenesis, progression, growth,differentiation and angiogenesis of gastric cancer. Low expression of PTEN can decrease expression of Caspase-3to disorder apoptosis of tumor cells, which might explain the molecular mechanisms of PTEN contributions to tumorigenesis and progression of gastric cancer.