circSMARCA5通过CCL5富集Treg细胞促进非小细胞肺癌细胞的增殖

目的：检测circSMRCA5在非小细胞肺癌（NSCLC）组织和细胞中的表达,以及其在NSCLC发生发展中的潜在功能和机制。方法：用qPCR 法检测circSMARCA5在NSCLC 组织中的表达。使用慢病毒转染法将circSMARCA5过表达质粒和对照质粒pLC5分别转染人肺癌A549 和H1975 细胞。采用qPCR法检测稳定转染细胞中circSMARCA5的表达水平。通过CCK-8、克隆形成、细胞周期和异种移植瘤实验检测circSMARCA5 过表达对A549 和H1975 细胞生物学行为的影响。通过转录组测序、KEGG和GO富集分析,确定circSMARCA5可能的靶基因。分别构建circSMARCA5过表达A549、Lewis 细胞BABL/c 裸鼠和免疫正常的C57 小鼠皮下移植瘤模型,观察circSMARCA5对裸鼠皮下移植瘤生长的影响,流式细胞术检测对Lewis 细胞移植瘤组织中Treg 细胞水平的影响。结果：circSMARCA5在NSCLC 组织中呈高表达（P<0.01）。过表达circSMARCA5可以在体外促进NSCLC 细胞的增殖（P<0.05,P<0.01）。体内实验中,circSMARCA5 可以促进裸鼠皮下移植瘤的生长（P<0.01）。机制上,经KEGG 和GO 富集分析,确定C-C 趋化因子配体5（CCL5）为circSMARCA5 的下游靶基因。过表达circSMARCA5 组A549 和H1975 细胞中CCL5 的表达量增加（均P<0.05）。circSMARCA5 介导的CCL5 上调促进了免疫正常的C57 小鼠皮下移植瘤的生长。C57 小鼠皮下移植瘤制备成的单细胞悬液行流式细胞术检测显示,circSMARCA5过表达组的Treg 细胞比例高于对照组[（3.1±0.5）% vs （1.0±0.1）%,P<0.05]。结论：circSMARCA5在NSCLC组织中呈高表达,其可能通过CCL5将Treg 细胞招募到肿瘤中,导致肿瘤的免疫逃逸,促进NSCLC的进展。     

miR-486-5p对裸鼠胃癌移植瘤生长的抑制作用

目的:探讨miR-486-5P对裸鼠皮下人胃癌移植瘤生长的影响及其可能的作用机制.方法:建立胃癌SGC-7901细胞裸鼠皮下移植瘤模型,经miR-486-5P过表达质粒处理后,观察裸鼠皮下移植瘤生长情况,采用Western blotting及免疫组织化学法检测miR-486-5p靶基因神经纤毛蛋白-2(NRP2)的表达水平.结果:成功构建胃癌SGC-7901细胞裸鼠皮下移植瘤模型;实验组移植瘤内miR-486-5p的表达水平明显高于对照组(P＜0.05),miR-486-5p可明显抑制裸鼠皮下胃癌移植瘤的生长;与阴性及空白对照组相比,实验组平均瘤体质量明显下降[(0.404±0.080) vs (0.748±0.122)、(0.788±0.176)g,均P＜0.05];移植瘤平均体积明显减小[(0.333 ±0.039) vs (0.597 ±0.175)、(0.594±0.216)cm3,均P＜0.05],实验组的抑瘤率达46.99％;实验组经miR-486-5p过表达质粒处理后,免疫组化检测结果显示,NRP2蛋白主要定位于胃癌细胞质内,呈棕黄色颗粒;实验组NRP2蛋白的IHS得分显著低于阴性及空白对照组[(2.2±0.84) vs (6.4±0.89),(6.2±1.48),均P＜0.01];阴性对照组与空白对照组的得分差异无统计学意义(P＞0.05);Western blotting检测结果显示,实验组移植瘤组织中NRP2蛋白的相对表达量显著低于阴性及空白对照组[(0.04±0.006) vs (0.70 ±0.03),(0.68±0.02),P＜0.01].阴性和空白对照组间的IHS得分值、抑瘤率及NRP2蛋白表达水平的差异均无统计学意义(P＞0.05).结论:miR-486-5p可明显抑制裸鼠皮下胃癌SGC-7901细胞移植瘤的生长,其作用机制可能与抑制NRP2的表达有关.     

lncRNA ZFAS1在肺癌中的表达及其对肺癌细胞生物学功能的影响及机制研究

目的:探究lncRNA ZFAS1在肺癌中的表达及其对肺癌细胞生物学功能的影响,同时通过构建肺癌裸鼠移植瘤模型,探寻其可能存在的分子机制。方法:采用qRT-PCR检测ZFAS1在肺癌组织和细胞系中的mRNA表达水平及荧光原位杂交(FISH)技术检测肺癌组织中ZFAS1的阳性指标;受试者工作特征(ROC)曲线检测血清中ZFAS1在肺癌临床诊断中的灵敏度和特异度;Transwell侵袭实验、细胞划痕实验及Tunel细胞凋亡实验检测上调/下调ZFAS1对NCI-H1299和NCI-H460细胞侵袭、迁移及凋亡的影响;检测肺癌裸鼠皮下移植瘤生长情况;Ki67和PCNA免疫荧光和PCNA免疫组化实验检测裸鼠体内细胞增殖情况。结果:ZFAS1 mRNA表达在肺癌组织和各细胞系中均上调;ROC曲线显示当ZFAS1的相对表达量取值为0.84时,其敏感度和特异度分别为86.7%和76.7%;过表达ZFAS1促进NCI-H1299细胞侵袭、迁移及抑制细胞凋亡,而干扰ZFAS1则抑制NCI-H460细胞侵袭、迁移及促进细胞凋亡;ZFAS1促进裸鼠皮下瘤体生长;ZFAS1在裸鼠皮下肺癌组织中的阳性指标显著增加...     

姜黄素下调HIF-1α/MMP-9通路抑制人胃癌裸鼠移植瘤生长

目的 研究姜黄素对人胃癌细胞株BGC-823裸鼠皮下移植瘤的生长抑制作用及其机制.方法 以0、1、5、10、20、50mg/L的姜黄素处理BGC-823细胞24或48 h,甲基噻唑盐(MTT)法及Transwell 分析细胞增殖和侵袭能力;荷瘤裸鼠检测姜黄素对肿瘤生长影响,同时Western blot检测瘤体内HIF-1α、MMP-9蛋白的表达;荧光素酶报告基因系统检测姜黄素对HIF-1α及HIF-1α对MMP-9启动子区域调控.结果 姜黄素处理人胃癌细胞BGC-823后,在体内体外的生长均受到抑制且作用呈量效依赖关系;细胞侵袭能力明显降低;HIF-1α、MMP-9蛋白的表达降低.姜黄素通过转录水平抑制HIF-1α及MMP-9的启动子活性.结论 姜黄素在体内外均抑制胃癌的生长,它可以通过降低HIF-1α活性,下调MMP-9水平,从而抑制肿瘤侵袭,发挥抗肿瘤作用.     

二甲双胍对甲状腺未分化癌细胞增殖和能量代谢的影响

目的:探讨二甲双胍对甲状腺未分化癌（anaplastic thyroid cancer,ATC）细胞增殖及能量代谢的影响,并检测糖酵解途径相关蛋白GLUT1、PKM2、LDHA表达的变化。方法:体外培养人ATC细胞和人正常甲状腺细胞,分为对照组和不同浓度二甲双胍（1、5、10mmol/L）处理组,并分别培养24h、48h。四甲基偶氮唑蓝（MTT）法检测细胞增殖,Seahorse能量代谢分析仪检测细胞糖酵解及线粒体呼吸的变化。Western blot检测各组细胞GLUT1、PKM2和LDHA蛋白表达变化。结果:与对照组相比,二甲双胍可抑制ATC细胞增殖,且呈剂量-时间依赖性（P<0.05）。二甲双胍可改变ATC细胞能量代谢模式,随二甲双胍处理浓度增加和时间延长,糖酵解和线粒体呼吸水平均可出现抑制,呈剂量-时间依赖性（P<0.05）。GLUT1、PKM2和LDHA在ATC细胞中过表达,而二甲双胍对ATC细胞中GLUT1、PKM2和LDHA蛋白的表达无明显影响。结论:二甲双胍可抑制ATC细胞增殖和能量代谢,但其机制仍有待于进一步研究。     

姜黄素下调HIF-1α/MMP-9通路抑制人胃癌裸鼠移植瘤生长北大核心CSCD

目的研究姜黄素对人胃癌细胞株BGC-823裸鼠皮下移植瘤的生长抑制作用及其机制。方法以0、1、5、10、20、50mg/L的姜黄素处理BGC-823细胞24或48h,甲基噻唑盐(MTT)法及Transwell分析细胞增殖和侵袭能力;荷瘤裸鼠检测姜黄素对肿瘤生长影响,同时Western blot检测瘤体内HIF-1α、MMP-9蛋白的表达;荧光素酶报告基因系统检测姜黄素对HIF-1α及HIF-1α对MMP-9启动子区域调控。结果姜黄素处理人胃癌细胞BGC-823后,在体内体外的生长均受到抑制且作用呈量效依赖关系;细胞侵袭能力明显降低;HIF-1α、MMP-9蛋白的表达降低。姜黄素通过转录水平抑制HIF-1α及MMP-9的启动子活性。结论姜黄素在体内外均抑制胃癌的生长,它可以通过降低HIF-1α活性,下调MMP-9水平,从而抑制肿瘤侵袭,发挥抗肿瘤作用。     

羟基磷灰石纳米粒子抑制胃癌细胞生长的体内外实验研究

目的:观察羟基磷灰石纳米粒子(Hydroxyapatite nanoparticles,Nano-HAP)对胃癌细胞系SGC-7901、BGC-823、MKN-28体外生长的影响及其对SGC-7901细胞皮下移植瘤裸鼠的抗瘤作用和药物毒性.方法:以不同浓度的Nano-HAP分别作用三种胃癌细胞48小时,MTT法检测其对胃癌细胞增殖的影响;以细胞划痕实验、黏附测定观察Nano-HAP对胃癌细胞侵袭力的影响;建立SGC-7901细胞皮下移植瘤裸鼠模型30只,随机分为对照组、Nano-HAP组和5-Fu组,观察每组移植瘤生长情况.结果:Nano-HAP可明显抑制胃癌细胞的增殖和生长,且呈剂量效应关系,并能降低胃癌细胞的体外侵袭和运动能力 (P＜0.05);同时可显著抑制SGC-7901细胞皮下移植瘤的生长,Nano-HAP组肿瘤体积明显小于对照组(P＜0.05),肿瘤生长抑制率达47.96%;5-Fu组抑瘤率达54.30%,但表现出明显不良反应.结论:Nano-HAP在体外和体内均可显著抑制胃癌细胞生长,并降低胃癌细胞体外侵袭和运动能力.     

miR-875-5p通过靶向USF2抑制胃癌细胞的增殖、迁移和侵袭

目的:探讨miR-875-5p对胃癌细胞增殖、迁移和侵袭的影响及其机制。方法:采用qPCR法检测胃癌细胞BGC-823、HGC-27、MGC-803、SGC-7901、AGS、MKN-45和胃黏膜上皮细胞GES-1中miR-875-5p的表达水平。利用脂质体转染技术,分别将miR-875-5p模拟物/抑制剂(mimic/inhibitor)及其阴性对照质粒(miR-NC/Anti-miR-NC)转染至AGS细胞/MKN-45细胞,构建过表达/抑制miR-875-5p的细胞模型,空白对照组(Control组)不转染。通过CCK-8、克隆形成、Transwell等实验分别检测miR-875-5p表达变化对细胞增殖、克隆形成、迁移和侵袭的影响。采用双荧光素酶报告基因实验验证miR-875-5p与上游刺激因子2(USF2)的靶向关系,WB实验验证miR-875-5p对USF2的调控作用并检测USF2蛋白的表达。构建MKN-45细胞裸鼠移植瘤模型,验证miR-875-5p过表达对MKN-45细胞成瘤能力的影响。结果:miR-875-5p在6种胃癌细胞中表达水平显著低于胃黏膜上皮细胞GES-1(均P<0.01)。与Control组和miR-NC组相比,miR-875-5p mimic组AGS细胞的增殖、克隆形成率、迁移和侵袭细胞数,以及USF2蛋白的表达均显著降低(P<0.05或P<0.01);miR-875-5p inhibitor组MKN-45细胞的增殖、克隆形成率、迁移和侵袭细胞数,以及USF2蛋白的表达均显著提高(P<0.05或P<0.01)。双荧光素酶报告基因实验证明,miR-875-5p能够直接靶向USF2基因。体内成瘤实验结果表明,过表达miR-875-5p显著抑制MKN-45细胞移植瘤的生长(均P<0.01)。结论:miR-875-5p通过靶向USF2抑制胃癌细胞的增殖、迁移和侵袭。     

tRF-Pro-CGG对小鼠胰腺癌细胞生物学行为的影响及其分子机制

背景与目的:tRNA衍生的片段(tRNA-derived fragments,tRF)是一类长度为14～30 nt的小分子非编码RNA,其影响着恶性肿瘤的发展进程。本研究旨在探讨tRF-Pro-CGG对小鼠胰腺癌细胞生物学行为的影响及其可能的分子机制。方法:采用实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RTFQ-PCR)检测tRF-Pro-CGG在小鼠胰腺癌细胞系pan02、LTPA,人胰腺癌细胞系Capan-2和正常胰腺细胞HPDE6-C7中的表达水平。通过慢病毒转染技术过表达pan02细胞及敲低LTPA细胞中tRF-Pro-CGG的表达,采用RTFQ-PCR和蛋白质印迹法(Western blot)检测过表达和敲低效果。采用细胞计数试剂盒-8(cell counting kit-8,CCK-8)检测细胞增殖情况。采用transwell实验检测细胞迁移和侵袭能力。采用动物模型检测tRF-Pro-CGG对胰腺癌裸鼠移植瘤生长和转移的影响。采用H-E染色观察移植瘤的组织病理学结构。...     

Lnc-BM通过FASTK/MT-ND6轴调节线粒体呼吸功能促进胃癌进展

目的 探究Lnc-BM在胃癌发生发展中的作用及分子机制。方法 收集36例胃癌患者的胃癌组织及配对的癌旁正常组织,RT-qPCR检测胃癌组织和癌旁正常组织中Lnc-BM的表达水平。构建Lnc-BM过表达或敲低细胞,平板克隆形成实验和CCK-8实验分析Lnc-BM对胃癌细胞增殖能力的影响,Transwell实验分析Lnc-BM对胃癌细胞迁移和侵袭能力的影响。RNA Pull-down实验分析Lnc-BM与FASTK蛋白的结合,Western blot检测Lnc-BM的表达对FASTK蛋白水平的影响。Western blot实验和Seahorse细胞能量代谢分析检测Lnc-BM过表达或敲降的胃癌细胞中线粒体相关蛋白的表达变化以及线粒体能量代谢的改变。裸鼠荷瘤实验进一步观察Lnc-BM对胃癌细胞体内生长的影响。结果 Lnc-BM在胃癌组织中表达明显高于对应的癌旁正常组织。实验结果显示,过表达Lnc-BM促进胃癌细胞增殖、迁移和侵袭,敲低Lnc-BM抑制胃癌细胞增殖、迁移和侵袭(均P＜0.05)。RNA Pull-down结果显示,Lnc-BM可直接结合FASTK蛋白。Western blot实...     

H19 promotes aerobic glycolysis,proliferation, and immune escape of gastric cancer cells through the microRNA-519d-3p/lactate dehydrogenase A axis

Long noncoding RNAs (lncRNAs) have been investigated in multiple human cancers including gastric cancer (GC). Our research aims to explore the role of H19 in aerobic glycolysis, proliferation, and immune escape of GC cells. The expression of H19 in GC samples was analyzed using Gene Expression Profiling Interactive Analysis, Gene Expression Omnibus data, and real-time quantitative PCR analysis. Relative quantification of glucose consumption and lactate production from cell supernatant were applied to assess the aerobic glycolysis of GC cells. Subcellular fractionation, luciferase reporter, and western blot assays certified the binding between genes. Cell Counting Kit-8 and colony formation assays were used to determine GC cell proliferation. Flow cytometry, ELISA, and real-time quantitative PCR assays were applied to analyze the immunosuppressive effect of H19. H19 was highly expressed in samples of patients with GC, and associated with tumor growth in vivo. H19 knockdown suppressed glucose consumption, lactate production, and proliferation of GC cells by regulating the microRNA (miR)-519d-3p/lactate dehydrogenase A (LDHA) axis. Both miR-519d-3p depletion and LDHA overexpression could reverse the H19 knockdown-induced decrease in aerobic glycolysis and proliferation. Moreover, conditioned medium from stable knockdown H19 GC cells modulated the activity of immune cells including γδT cells, Jurkat cells, and tumor-associated macrophages in a miR-519d-3p/LDHA/lactate axis-dependent manner. The H19/miR-519d-3p/LDHA axis mainly contributed to aerobic glycolysis, proliferation, and immune escape of GC cells.     

微小RNA-106 a靶向PTEN通路对胃癌细胞的增殖与凋亡的调控作用

目的 探讨微小RNA-106a(miR-106a)靶向磷酸酶和张力蛋白同源物(PTEN)通路对胃癌细胞的增殖与凋亡的调控作用.方法 对数生长期的胃癌BGC-823细胞为三组:miR-106a组、对照组与空白组,分别转染miR-106a mimic、miR-NC与等体系的磷酸盐缓冲液.采用CCK法检测细胞增殖,流式细胞仪...     

circPVT1 promotes osteosarcoma glycolysis and metastasis by sponging miR-423-5p to activate Wnt5a/Ror2 signaling

Osteosarcoma (OS) is the most prevalent form of bone cancer. It has a high metastatic potential and progresses rapidly. The molecular mechanisms of OS remain unclear and this study aims to examine the functional role of circPVT1 and miR-423-5p in OS. Quantitative RT-PCR (qRT-PCR) and western blotting were used to examine levels of miR-423-5p, circPVT1, Wnt5a, Ror2, and glycolysis-related proteins, including HK2, PKM2, GLUT1, and LDHA. Colony formation and transwell assays were used to test the roles of miR-423-5p, circPVT1, and Wnt5a/Ror2 in OS cell proliferation, migration, and invasion. Dual luciferase assay and Ago2-RIP were used to validate the interactions of miR-423-5p/Wnt5a, miR-423-5p/Ror2, and circPVT1/miR-423-5p. Glucose uptake assay and measurement of lactate production were performed to assess the glycolysis process. A nude mouse xenograft model was used to evaluate the effects of sh-circPVT1 and miR-423-5p mimics on tumor growth and metastasis in vivo. miR-423-5p was reduced in both OS tissues and OS cell lines, while Wnt5a/Ror2 and circPVT1 were elevated. miR-423-5p bound to 3′-UTR of Wnt5a and Ror2 mRNA, and inhibited glycolysis and OS cell proliferation, migration, and invasion by targeting Wnt5a and Ror2. circPVT1 interacted with miR-423-5p and activated Wnt5a/Ror2 signaling by sponging miR-423-5p. Knockdown of circPVT1 or overexpression of miR-423-5p suppressed OS tumor growth and metastasis in vivo. miR-423-5p inhibited OS glycolysis, proliferation, migration, and metastasis by targeting and suppressing Wnt5a/Ror2 signaling pathway, while circPVT1 promoted those processes by acting as a sponge of miR-423-5p.     

LncRNA ANCR调控miR-331表达影响胃癌细胞的生物学行为

背景与目的：胃癌是导致癌症死亡的第二大原因，是东亚、东欧及中南美洲部分地区最常见的胃肠道恶性肿瘤。该研究旨在探讨长链非编码RNA（long-chain noncoding RNA，lncRNA）抗分化非编码RNA（antidifferentiation noncoding RNA，ANCR）靶向miR-331调节胃癌细胞的增殖、侵袭行为及其机制。方法：河南省肿瘤医院2016年1月1日—2016年12月1日获得60例临床胃癌样本和20例对应的癌旁组织标本（距癌组织2 cm）。实时荧光定量聚合酶链反应（real-time fluorescence quantitative polymerase chain reaction，RTFQ-PCR）检测ANCR在胃癌组织和不同胃癌细胞株中的表达情况；分析ANCR的表达和胃癌患者临床病理学特征之间的关系；平板克隆实验检测ANCR对胃癌细胞增殖能力的影响；Transwell侵袭实验检测ANCR对胃癌细胞侵袭能力的影响；裸鼠成瘤实验检测ANCR对胃癌细胞肿瘤异种移植的影响；双荧光素酶报告基因检测ANCR与miR-331之间的相互作用。结果：胃癌组织中ANCR的表达（4.18±0.28）高于正常组织（1.45±0.15），差异有统计学意义（t=12.36，P=0.045）；与其他胃癌细胞株相比，BGC-823和MGC-803细胞中ANCR的表达水平较高（9.12±0.52），差异有统计学意义（P=0.019）；抑制ANCR的表达可以减弱胃癌细胞的增殖能力（98.8±10.4）和侵袭能力（175.9±5.7）；抑制ANCR的表达后胃癌细胞在裸鼠体内异种移植能力（223.4±13.4）弱于NC组（115.6±10.7），差异有统计学意义（t=13.28，P=0.014）；双荧光素酶实验证实ANCR可以直接调控miR-331的表达及荧光活性。结论：ANCR可以靶向调节miR-331的表达影响胃癌细胞的增殖、侵袭行为，影响胃癌细胞在裸鼠体内的生长情况。     

miR-323a-3p通过靶向结合TM4SF1而调控NSCLC A549细胞的增殖、迁移和侵袭

目的:探究miR-323a-3p、四次穿膜蛋白超家族成员1(TM4SF1)在NSCLC组织和细胞中的表达及两者间的靶向调控关系,观察两者表达对A549细胞增殖、迁移、侵袭和裸鼠移植瘤生长的影响。方法:收集2014年1月至12月间青海省人民医院手术切除的20例NSCLC组织及其相应的癌旁组织,qPCR和WB法检测癌组织中miR-323a-3p、TM4SF1 mRNA和TM4SF1蛋白的表达。向A549细胞转染miR-323a-3p mimic,采用MTT法、Transwell法、WB法检测miR-323a-3p过表达对细胞的增殖、迁移和侵袭以及TM4SF1、细胞周期蛋白D1(cyclin D1)、p21、MMP-2、MMP-9蛋白表达的影响。采用生物信息学预测工具StarBase和双荧光素酶报告基因实验分析miR-323a-3p与TM4SF1靶向关系。将si-TM4SF1转染至A549细胞,以及分别将miR-323a-3p mimic与pcDNA或pcDNA-TM4SF1共转染A549细胞,评估细胞增殖、迁移和侵袭能力的变化;同时建立各组细胞的BALB/c裸鼠移植瘤模型,在14、21和2...     

microRNA-21通过Bcl-2通路调节胃癌紫杉醇化疗耐药性

目的:研究miR-21在胃癌组织中的表达水平及对胃癌生物学行为的影响,探讨miR-21与紫杉醇化疗耐药性的关系及机制。方法:实时荧光定量（real-time quantitative polymerase chain reaction,RT-qPCR）比较25例胃癌组织和癌旁组织中miR-21的表达差异;细胞实验探讨miR-21对胃癌细胞增殖、凋亡能力的影响;MTT法测定紫杉醇对SGC-7901细胞的生长抑制率;Western blot验证Bcl-2、Bax、PTEN蛋白表达和miR-21的关系。结果:miR-21在胃癌组织中表达水平高于癌旁正常组织（P=0.015）;与NC组比较,转染miR-21模拟物的胃癌SGC-7901细胞增殖能力（P＜0.05）增强,下调miR-21表达水平可促进胃癌细胞凋亡(P＜0.05);转染miR-21 mimic组的紫杉醇对胃癌细胞的抑制率明显低于miR-21 inhibitor组;miR-21 inhibitor组较NC组中Bcl-2的蛋白水平下降（P=0.000）,而PTEN（P=0.039）、Bax（P=0.037）表达升高。结论:miR-21在胃癌组织中过表达,且过表达的miR-21可能通过升高Bcl-2表达并降低Bax、PTEN蛋白表达从而促进胃癌细胞增殖,抑制胃癌细胞凋亡,并增加紫杉醇化疗耐药性。     

p53/Lactate dehydrogenase A axis negatively regulates aerobic glycolysis and tumor progression in breast cancer expressing wild‐type p53

Tumor suppressor p53 is a master regulator of apoptosis and plays key roles in cell cycle checkpoints. p53 responds to metabolic changes and alters metabolism through several mechanisms in cancer. Lactate dehydrogenase A (LDHA), a key enzyme in glycolysis, is highly expressed in a variety of tumors and catalyzes pyruvate to lactate. In the present study, we first analyzed the association and clinical significance of p53 and LDHA in breast cancer expressing wild‐type p53 (wt‐p53) and found that LDHA mRNA levels are negatively correlated with wt‐p53 but not with mutation p53 mRNA levels, and low p53 and high LDHA expression are significantly associated with poor overall survival rates. Furthermore, p53 negatively regulates LDHA expression by directly binding its promoter region. Moreover, a series of LDHA gain‐of‐function and rescore experiments were carried out in breast cancer MCF7 cells expressing endogenous wt‐p53, showing that ectopic expression of p53 decreases aerobic glycolysis, cell proliferation, migration, invasion and tumor formation of breast cancer cells and that restoration of the expression of LDHA in p53‐overexpressing cells could abolish the suppressive effect of p53 on aerobic glycolysis and other malignant phenotypes. In conclusion, our findings showed that repression of LDHA induced by wt‐p53 blocks tumor growth and invasion through downregulation of aerobic glycolysis in breast cancer, providing new insights into the mechanism by which p53 contributes to the development and progression of breast cancer.     

反义microRNA-221与反义microRNA-222抑制人脑胶质瘤细胞的体外与体内生长

目的 探讨敲低microRNA(miR)-221和miR-222表达以抑制人脑胶质瘤U251细胞生长的作用及其机制.方法 脂质体介导转染反义寡聚核苷酸(AS-miR-221和AS-miR-222)于人脑胶质瘤细胞U251.采用Northern blot鉴定转染后U251细胞的miR-221和miR-222表达水平;四甲基偶氮唑蓝(MTT)法评价AS-miR-221和AS-miR-222抑制U251细胞生长的作用;Transwell实验检测细胞侵袭能力;流式细胞术检测细胞周期的分布和凋亡;Western blot检测转染后U251细胞相关蛋白表达的变化,并用AS-miR-221和AS-miR-222治疗裸鼠皮下移植瘤,观察其在活体内对肿瘤生长的抑制作用.结果 Northern blot检测结果 显示,AS-miR-221和AS-miR-222共转染后,肿瘤细胞miR-221和miR-222表达明显下降,细胞生长速度降低,细胞穿过率为14.5%,细胞周期出现G0/G1期阻滞,凋亡率(13.7%)增高,并可见connexin43、p27、PUMA、caspase-3、PTEN、TIMP3和Bax等相关蛋白表达增高,而bcl-2表达降低,p53无明显变化.经AS-miR-221和AS-miR-222治疗后,裸鼠皮下移植瘤生长明显受抑.结论 AS-miR-221和AS-miR-222共转染可抑制U251细胞的增殖与侵袭,miR-221和miR-222可以作为人脑胶质瘤基因治疗的侯选靶点.     

microRNA-21 promotes tumor proliferation and invasion in gastric cancer by targeting PTEN

Gastric cancer is one of the most common carcinomas in China. microRNAs, a type of non-coding RNA, are important specific regulators and are involved in numerous bioprocesses of an organism. microRNA-21 (miR-21) has been identified as the most suitable choice for further investigation because it is overexpressed in nearly all solid tumors; furthermore, it has been demonstrated that miR-21 is involved in the genesis and progression of human cancer. It has been reported that PTEN, an important tumour suppressor, is regulated by multiple miRNAs. Thus, in this study we focused on the expression and significance of miR-21 in gastric cancer tissues, and the role of miR-21 in the biological behaviour and the expression of PTEN in gastric cancer cells. Real-time PCR was used to detect miR-21 expression in gastric cancer tissues, the adjacent normal tissues, and the gastric cell lines. The gastric cancer cell line BGC-823 was transfected with pre-miR-21/miR-21 inhibitor to overexpress/downregulate miR-21. The influence of miR-21 on the biological behaviour of gastric cancer cells was evaluated using the CCK-8 kit, FCMs, the scratch healing assay and the transwell test. Western blotting and the Luciferase Reporter Assay were used to evaluate the change of PTEN expression after lowered expression of miR-21 in gastric cancer cell lines. Real-time PCR analysis indicated that miR-21 exhibited higher expression in gastric cancer tissues compared to the adjacent non-tumor tissues. miR-21 expression was significantly associated with the degree of differentiation of the tumour tissues (P=0.004), as well as local invasion and lymph node metastasis (P<0.01). After transfection, pre-miR21 BGC-823 cells grew faster than the negative and control groups (P<0.01). The reduction in miR-21 expression demonstrated a remarkable effect on the biological behaviour of gastric cancer cells (P<0.05); the pre-miR-21-transfected cells healed more quickly compared to the control cells in the scratch healing assay, whereas the transwell test indicated that cell migration in vitro was notably inhibited with the downregulation of miR-21 (P<0.05). The western blot results and Luciferase Reporter Assay demonstrated that PTEN expression was remarkably increased after miR-21 inhibition (P<0.05). microRNA-21 expression was upregulated in gastric carcinoma tissues and was significantly associated with the degree of differentiation of tumour tissues, local invasion and lymph node metastasis. Overexpression of miR-21 promoted BGC-823 cell growth, invasion and cell migration in vitro, whereas downregulation of miR-21 exhibited a stronger inhibitory effect on the biological behaviour of gastric cancer cells; additionally, miR-21 inhibition may upregulate the PTEN expression level, which indicates that PTEN may be a target gene for gastric cancer initiation and development.     

miR-496通过SFMBT1抑制子宫颈癌HeLa细胞增殖、迁移和侵袭的相关研究

背景与目的:微小RNA(microRNA,miRNA)已被证实与子宫颈癌的发病相关.分析miR-496与SFMBT1的相互关系,阐明miR-496在子宫颈癌中的作用及其机制.方法:预测并验证miR-496的靶基因.实时荧光定量聚合酶链式反应(real-time fluorescence quantitative pol...