微小RNA在胃癌中的研究现状

微小RNA(miRNA)与胃癌具有密切关系.根据miRNA对胃癌促进和抑制的不同作用,可将miRNA分为癌基因和抑癌基因.由于miRNA在胃癌和健康人群中表达具有差异,因此可成为新型的肿瘤标志物进行胃癌辅助诊断和预测治疗反应及预后.掌握miRNA与胃癌耐药的关系,有利于研发新的治疗方式.本文将从miRNA与胃癌的发生发...     

Integrative analysis of mRNA and microRNA expression of a human alveolar epithelial cell(A549) exposed to water and organic‐soluble extract from particulate matter (PM)2.5

MicroRNA (miRNA) is now attracting attention as a powerful negative regulator of messenger RNA(mRNA) levels, and is implicated in the modulation of important mRNA networks involved in toxicity. In this study, we assessed the effects of particulate matter 2.5 (PM_2.5), one of the most significant air pollutants, on miRNA and target gene expression. We exposed human alveolar epithelial cell (A549) to two types of PM_2.5[water (W‐PM_2.5) and organic (O‐PM_2.5) soluble extracts] and performed miRNA microarray analysis. A total of 37 miRNAs and 62 miRNAs were altered 1.3‐fold in W‐PM_2.5 and O‐PM_2.5, respectively. Integrated analyses of miRNA and mRNA expression profiles identified negative correlations between miRNA and mRNA in both W‐PM_2.5 and O‐PM_2.5 exposure groups. Gene ontology and Kyoto encyclopedia of genes and genomes (KEGG) pathway analyses showed that the 35 W‐PM_2.5 target genes are involved in responses to nutrients, positive regulation of biosynthetic processes, positive regulation of nucleobase, nucleoside, and nucleotide, and nucleic acid metabolic processes; while the 69 O‐PM_2.5 target genes are involved in DNA replication, cell cycle processes, the M phase, and the cell cycle check point. We suggest that these target genes may play important roles in PM_2.5‐induced respiratory toxicity by miRNA regulation. These results demonstrate an integrated miRNA‐mRNA approach for identifying molecular events induced by environmental pollutants in an in vitro human model. © 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 302–310, 2017.     

二烯丙基三硫诱导人胃癌细胞凋亡相关基因的差异表达

目的探讨二烯丙基三硫(diallyl trisulfide,DATS)诱导人胃癌MGC803细胞凋亡的分子机制。方法MTT法检测DATS对MGC803细胞的生长抑制作用,荧光显微镜和流式细胞仪观察DATS作用后的细胞凋亡。人细胞凋亡基因芯片检测DATS作用MGC803细胞的凋亡相关基因表达谱,RT-PCR验证凋亡相关基因。结果MTT结果显示,4、8、12、16、24mg.L-1DATS对MGC803细胞生长的抑制率从11%增至78%,呈明显量效关系(P<0.05)。荧光显微镜下,DATS处理后的MGC803细胞核染色质着橘红色并呈固缩状或片段状。流式细胞仪检测显示,8、12、16、24mg.L-1DATS作用24h后,出现典型亚二倍体峰,平均凋亡率分别为11.4%、23.7%、27.4%、31.1%(P<0.05)。人细胞凋亡基因芯片检测结果表明,16mg.L-1的DATS作用4h后出现16个凋亡相关基因表达差异,RT-PCR证实,SODD基因mRNA表达上调、Apaf-1基因mRNA表达下调(P<0.05)。结论DATS诱导人胃癌MGC803细胞凋亡可能与多个基因和信号转导通路作用有关。     

斑贞1号、川芎嗪和氟尿嘧啶联合对人胃癌细胞多药耐药基因1表达的影响

目的探讨斑贞1号、川芎嗪（TMP）和氟尿嘧啶（5-FU）联合逆转人胃癌细胞系SGC-7901／ADR与多药耐药基因1（MDR1）表达的相关性。方法以SGC-7901／ADR细胞为靶细胞，以不加药组为阴性对照组，氟尿嘧啶组、斑贞1号组、斑贞1号＋氟尿嘧啶组、斑贞1号＋氟尿嘧啶＋川芎嗪（TMP）组为实验组，采用半定量RT-PCR法检测SGC-7901／ADR细胞在不同药物作用下MDR1mRNA的表达情况。结果氟尿嘧啶组与阴性对照组相比，MDR1mRNA表达差异无统计学意义（P〉0．05），而其他药物组与对照组及组间比较差异有统计学意义（P〈0．05），斑贞1号＋氟尿嘧啶＋TMP组表达量最少。结论斑贞1号、TMP和氟尿嘧啶联合逆转SGC-7901／ADR细胞耐药性可能与MDR1基因相关，为当前胃癌的治疗提供了新的理论依据。     

Iptkalim inhibits cocaine challenge—induced enhancement of dopamine levels in nucleus accumbens and striatum of rats by up—regulating Kir6.1 and Kir6.2 mRNA expression

目的:研究钾通道开放剂埃他卡林对急慢性可卡因应用引起的伏隔核、纹状体和额叶皮层的多巴胺和谷氨酸水平变化的影响及其机制。方法:采用高效液相色谱与电化学检测、荧光检测联用的方法测定各脑区谷氨酸和多巴胺的含量;采用半定量逆转录聚合酶链反应(RT-PCR)研究ATP敏感性钾通道亚单位Kir6.1、Kir6.2、SUR1和SUR2 mRNA表达的变化。结果:埃他卡林不影响急性可卡因应用引起纹状体和伏隔核中多巴胺和谷氨酸水平的增高(P>0.05),能够逆转激发剂量可卡因诱导的慢性成瘾大鼠纹状体和伏隔核的多巴胺含量增高(P<0.05),对激发后皮层和伏隔核谷氨酸水平增高有降低趋势但差异无显著性(P>0.05)。激发剂量可卡因能提高可卡因预处理组和埃他卡林预处理组纹状体和伏隔核的Kir6.1和Kir6.2 mRNA表达以及皮层的Kir6.2 mRNA表达,而且IPT预处理组的升高幅度显著高显著高于可卡因慢性处 理组。结论:埃他卡林通过上调Kir6.1和Kir6.2 mRNA表达抑制可卡因激发引起的纹状体和伏隔核的多巴胺水平的增高。     

MiR-497对人胃癌细胞株顺铂耐药性的影响和机制

目的:通过建立人胃癌细胞SGC-7901的顺铂耐药细胞株SGC-7901/DDP,探讨miR-497对SGC-7901/DDP耐药性的影响及其机制。方法:体外研究采用顺铂体外逐步加量诱导法建立人胃癌细胞SGC-7901耐药株,并通过检测药物半抑制浓度和耐药基因MDR1、BCRP、MRP1的表达以鉴定耐药细胞株;检测在亲本及耐药细胞中miR-497、MDR1、MRP1、BCRP和凋亡相关基因Bax、Bcl-2的表达水平;miR-497模拟物分别转染SGC-7901/DDP细胞株,利用SRB法和流式细胞术检测转染miR-497模拟物后细胞对顺铂醇的敏感程度和细胞的周期、凋亡的变化,并检测耐药基因和凋亡相关基因的表达。结果:成功建立人胃癌SGC-7901/DDP耐药细胞株,耐药细胞株中耐药基因MDR1、BCRP、MRP1表达均升高,抗凋亡基因Bcl-2升高,凋亡基因Bax下降,miR-497表达下降(P<0.05);miR-497模拟物提高耐药细胞株对顺铂的药物敏感性,凋亡水平增加,细胞周期G₀/G₁期细胞增多(P<0.05),并可抑制耐药细胞中耐药基因的表达,降低Bcl-2/Bax比值(P<0.05)。结论:人胃癌耐药细胞株SGC-7901/DDP的miR-497表达下调;上调miR-497的表达可逆转人胃癌SGC-7901/DDP细胞株对化疗药物顺铂的耐药性。     

MicroRNAs are involved in the development and progression of gastric cancer

MicroRNAs (miRNAs) are recognized as an essential component of the RNA family, exerting multiple and intricate biological functions, particularly in the process of tumorigenesis, proliferation, and metastatic progression. MiRNAs are altered in gastric cancer (GC), showing activity as both tumor suppressors and oncogenes, although their true roles have not been fully understood. This review will focus upon the recent advances of miRNA studies related to the regulatory mechanisms of gastric tumor cell proliferation, apoptosis, and cell cycle. We hope to provide an in-depth insight into the mechanistic role of miRNAs in GC development and progression. In particular, we summarize the latest studies relevant to miRNAs’ impact upon the epithelial-mesenchymal transition, tumor microenvironment, and chemoresistance in GC cells. We expect to elucidate the molecular mechanisms involving miRNAs for better understanding the etiology of GC, and facilitating the development of new treatment regimens for the treatment of GC.     

Molecular mechanisms and theranostic potential of miRNAs in drug resistance of gastric cancer

Introduction: Systemic chemotherapy is a curative approach to inhibit gastric cancer cells proliferation. Despite the great progress in anti-cancer treatment achieved during the last decades, drug resistance and treatment refractoriness still extensively persists. Recently, accumulating studies have highlighted the role of miRNAs in drug resistance of gastric cancers by modulating some drug resistance-related proteins and genes expression. Pre-clinical reports indicate that miRNAs might serve as ideal biomarkers and potential targets, thus holding great promise for developing targeted therapy and personalized treatment for the patients with gastric cancer.

Areas covered: This review provide a comprehensive overview of the current advances of miRNAs and molecular mechanisms underlying miRNA-mediated drug resistance in gastric cancer. We particularly focus on the potential values of drug resistance-related miRNAs as biomarkers and novel targets in gastric cancer therapy and envisage the future research developments of these miRNAs and challenges in translating the new findings into clinical applications.

Expert opinion: Although the concrete mechanisms of miRNAs in drug resistance of gastric cancer have not been fully clarified, miRNA may be a promising theranostic approach. Further studies are still needed to facilitate the clinical applications of miRNAs in drug resistant gastric cancer.     

人胃癌顺铂耐药细胞系的建立及其生物学特征

目的:国内外普遍应用体外建立的耐药细胞系作为研究模型。为探讨胃癌对顺铂耐药的机理,本研究首次建立人胃癌顺铂耐药细胞系并且研究其生物学特性。方法:采用逐步递增顺铂浓度,间歇作用体外诱导法建立人胃癌顺铂耐药细胞系SGC 7901/DDP;M TT法测定药物敏感性;光镜、电镜、M TT法活细胞计数、流式细胞仪及其相关P-糖蛋白的表达等方法观察其生物学特征的改变。结果:历时5个月建成人胃癌顺铂耐药细胞系SGC 7901/DDP,其对顺铂的耐药指数为14.68,并且与5-氟尿嘧啶、丝裂霉素等多种抗癌药有不同程度的交叉耐药性;SGC 7901/DDP的细胞形态及其P-糖蛋白的表达;体外群体倍增时间较亲代细胞延长;细胞周期分析发现其S期与G2/M期细胞减少、G0/G1期细胞增多。结论:SGC 7901/DDP细胞具有耐药特征,耐药性能稳定,与P-糖蛋白的表达有关。     

姜黄素对裸鼠人胃癌细胞 MMP-2和 CD44 v6在 mRNA水平表达的影响

目的：观察姜黄素对胃癌组织中MMP-2和CD44v6在mRNA水平表达的影响,探讨其抑制胃癌细胞转移的作用机制。方法以SGC-7901胃癌细胞株建立胃癌裸鼠原位移植瘤模型,将裸鼠随机分为对照组及姜黄素组,每组12只。观察裸小鼠胃癌种植后肿瘤生长及转移灶情况,用原位杂交方法检测肿瘤组织中MMP-2 mRNA和CD44 v6 mRNA表达的变化。结果所有荷瘤鼠胃壁均有肿瘤生长,荷瘤率100％,姜黄素组肝、腹腔和淋巴结的转移比对照组明显减少（ P ＜0ú．05）;姜黄素治疗组瘤体中MMP-2 mRNA和CD44v6 mRNA的表达明显下调（ P ＜0．05）。结论姜黄素具有抑制人胃癌裸鼠原位移植瘤转移的作用,其作用机制可能与下调MMP-2 mRNA和CD44 v6 mRNA的表达有关。     

二烯丙基二硫对小鼠肾包膜下移植人胃癌细胞的生长抑制作用

二烯丙基二硫(diallyl disulfide,DADS)是大蒜的主要有效成分,对多种肿瘤均有明显的抑制作用[1].本室体外研究表明:DADS可明显抑制人胃癌MGC803细胞生长[2],其机制与G2/M期阻滞和信号传导通路ERK/AP-1途径有关[3,4].本研究采用人肿瘤细胞肾包膜下移植模型,探讨DADS对体内生长的MGC803细胞的抑制作用.     

snoRNA 表达谱与胃癌预后的关联研究

Abstract:Objective To identify snoRNA, which may be related to prognosis of gastric cancer. Methods Ninetygastric cancer patients who diagnosed at Tianjin Medical University Cancer Institute and Hospital were randomly collected in this study, and their clinical data were followed up. A total of 405 snoRNA expression profiles were analyzed in 90 gastric cancer patients. Patients were classified as "low expression" group or "high expression" group according to the median expression of each snoRNA expression, which was calculated by univariate and multivariate survival analysis. We also screened out the snoRNAs, in which patients were survived differently. Patients were classified as high, middle, or low risk groups based on the snoRNA risk score. Values of age, gender, smoking, drinking, histological differentiation (well, moderately-differentiated and poorly differentiated), clinical stage (Ⅰ+Ⅱstage and Ⅲ+Ⅳstage), tumor size (<5 cm and ≥5 cm), tumor location (upper 1/3 and others) and snoRNA isk score (high, middle, and low risk group) were assessed by multivariate Cox analysis. Results There were significant differences in overall survival and (or) progression-free survival rates in 19 patients with high and low snoRNAs expressions (P < 0.05). Results of multivariate Cox analysis showed that patients with high expression of ACA61, ACA27 and U36A showed a higher overall survival and progression-free survival rates, while patients with high expression of ENSG00000206898 showed a lower overall survival and progression-free survival survival rates (P < 0.01). SnoRNA risk score is an independent prognostic factor or patients with gastric cancer. Compared with low risk group, patients in middle risk group and in high risk group showed a shorter overall survival and progression-free survival rates (P < 0.001). Conclusion The expressions of ACA61, ACA27, U36A and ENSG00000206898 are independent prognostic factors of gastric cancer. Low expressions of the first three indexes and high expressions of the last one predict a bad prognosis.     

原核移植技术对子代小鼠脑组织mRNA表达谱的影响

目的 探讨原核移植（PNT）技术对子代小鼠脑组织 mRNA 表达谱的影响。 方法 将 SPF 级 ICR（CD-1）雌鼠超促排卵并与雄鼠交配受精后, 收集雌鼠受精卵进行原核移植, 获得重构胚胎, 将其移植到假孕小鼠输卵管内获得原核移植子代小鼠（PNT 组）, 未经原核移植操作的受精卵经过胚胎移植后获得子代小鼠（对照组）。 提取 2 组小鼠脑组织 RNA, 逆转录为 cDNA 后荧光染料标记, 利用 Agilent 小鼠 mRNA 芯片检测 2 组 mRNA 表达谱差异, 筛选差异表达的 mRNA 并进行 GO 和信号通路分析。 结果 PNT 组与对照组表达差异倍数在 2 倍以上的 mRNAs 有 392 个, 占所有 mRNAs 的 1.7%, 其中 366 个表达升高, 26 个表达降低;表达差异倍数在 4 倍以上的共 11 个。 上述差异表达的 mRNAs 经 GO 分析结果显示靶基因富集于 mRNA 可变剪接、小 GTP 酶介导的信号转导、胰岛素受体信号通路调节等生物学过程以及水解酶活性、跨膜转运蛋白活性、焦磷酸酶活性等分子功能。 信号通路富集分析结果显示靶基因集中在离子通道转运、脂肪酸代谢、丁酸甲酯代谢、三酰甘油和酮体代谢等信号通路。 结论 原核移植可能会对小鼠脑组织一些关键代谢过程产生影响。     

多西他赛联用替吉奥或卡培他滨治疗胃癌的成本—效果分析

目的比较替吉奥与卡培他滨分别联合多西他赛(TXT)治疗进展期胃癌的有效性、安全性及经济效果。方法试验分A、B两组,第1天均给予多西他赛75 mg/m2,静脉滴注2 h;第1～14天,A组每天给予卡培他滨2 000 mg/m2,B组每天给予替吉奥胶囊60 mg/m2,两组给药均分为两等份于早晚餐后0.5 h用水吞服,服用2周后休息1周,3周为1个周期,至少完成2个周期,最多完成6个周期后评价。结果 50例均可评价疗效。A、B两组的总有效率分别为46.21%和50.00%,疾病控制率为76.92%和75.00%(P>0.05),平均成本分别为17 158.90元和10 094.37元。成本–效果比分别为371.81和201.89。两组不良反应主要包括骨髓抑制、恶心呕吐、腹泻、口腔黏膜炎和手足综合征等,以1～2级为主,均可耐受。结论两组治疗晚期胃癌的疗效相当,不良反应均可耐受,但B组成本–效果明显低于A组,因此替吉奥联合多西他赛治疗进展期胃癌方法更优,值得临床推广。     

胃部分切除手术治疗早期胃癌的疗效分析

目的探讨保留幽门与迷走神经的胃部分切除术治疗早期胃癌的临床价值。方法选取本院2009年3月-2010年3月本院普通外科收治的早期胃癌患者140例,根据临床术式不同分为观察组与对照组各70例,对照组行远端胃切除术,观察组行保留迷走神经与幽门的胃部分切除术,对比两组的临床疗效。结果观察组患者服药15、30、60min的胃排空功能相比对照组均显著改善（P〈0．05）;胆囊收缩功能仅在60min后明显高于对照组（P〈O．05）,15min与30min两组相比差异无统计学意义（P〉0．05）。两组3年的生存率与胃癌复发率相比差异无统计学意义（P〉0．05）。结论保留幽门与迷走神经的胃部分切除术治疗早期胃癌能够有效改善患者术后胃排空与胆囊收缩功能,且术后BMI指数明显提升,是胃癌早期手术中临床价值较高的术式。     

RNA干扰对5637膀胱癌细胞生存素基因表达的影响

目的 研究RNA干扰（RNAi）对膀胱癌5637细胞生存素（survivin）基因表达的干扰效应。方法 化学合成生存素序列特异性双链RNA（dsRNA）,用Lipofectamine 2000转染5637细胞,采用荧光定量PCR检测生存素基因的表达,用流式细胞仪检测Annexinv标记的凋亡细胞。结果 序列特异性dsRNA-生存素可显著抑制生存素基因在转录水平上的表达,和阴性对照组相比,浓度为40nmol／L的dsRNA-生存素在转染5637细胞24、48h后生存素mRNA表达的抑制率分别为29％和53％,浓度为100nmol／L时抑制率为46％和86％。生存素基因表达的抑制与5637细胞的凋亡呈正相关。结论 转染dsRNA-生存素可明显抑制膀胱癌5637细胞株生存素基因的表达,并伴有细胞凋亡。     

Microarray analysis of altered long non-coding RNA expression profile in liver cancer cells treated by ginsenoside Rh2

Microarray expression profiles of lncRNAs and mRNAs were investigated in HepG2 cells treated with 20?μg/ml ginsenoside Rh2 as well as in ginsenoside Rh2-untreated cells. Microarray analysis showed 618 upregulated lncRNAs and 161 downregulated lncRNAs in HepG2 cells treated with ginsenoside Rh2 compared with the control group. Moreover, three differentially expressed lncRNAs were validated by quantitative real-time polymerase chain reaction (qRT-PCR). This may be beneficial to patients as an anti-cancer treatment and potentially provide novel targets for HCC (hepatocellular carcinoma) therapy.     

人乳腺珠蛋白和亲脂素B共转染乳腺癌细胞T47-D

目的构建人乳腺珠蛋白（Mammaglobin A）和亲脂素B（Lipophilin B）真核表达载体,建立稳定高表达Mammaglobin A和同时稳定高表达Mammaglobin A和Lipophilin B的人乳腺癌T47-D细胞系。方法通过RT-PCR的方法从人乳腺癌细胞系MDA—MB-415中扩增MammaglobinA和LipophilinB基因,构建真核表达载体pc—MA、pc—LP,分别独立及共转染T47-D细胞,G418筛选阳性克隆,并利用RT—PCR、Western Blot鉴定。结果重组质粒pc—MA、pc—LP酶切图谱、序列分析证明pc—MA、pc—LP载体构建成功,目的基因序列与GenBank中的序列完全一致。RT-PCR与Western Blot结果证实,独立稳定高表达Mammaglobin A和同时稳定高表达Mammaglobin A和Lipophilin B的人乳腺癌细胞系建立成功。结论成功建立独立高表达Mammaglobin A和同时高表达Mammaglobin A和Lipophilin B的乳腺癌细胞系,为探讨Maremaglobin A和Lipophilin B在乳腺癌发生发展中的作用提供了实验基础。