微小RNA-378a对皮肤鳞癌细胞侵袭、迁移和CD226表达的影响

目的探讨微小RNA-378a(miR-378a)对皮肤鳞癌(CSCC)细胞侵袭、迁移和CD226表达的影响。方法采用实时荧光定量PCR(QPCR)检测CSCC细胞A431和人角质形成细胞HaCaT的miR-378a水平;将体外常规培养的A431细胞进行转染,根据转染物不同分为增强转染组(转染miR-378a mimics)、抑制转染组(转染miR-378a inhibitor)和空白转染组(无转染)。采用QPCR、活细胞计数(CCK)-8、AnnexinⅤ-FITC/PI双染流式细胞术、Transwell细胞迁移和侵袭实验以及Western blotting检测miR-378a水平、增殖活力、凋亡率、穿膜细胞数和CD226水平;构建荧光素酶报告质粒psiCHECK-2,并采用双荧光素酶报告基因实验验证miR-378a与CD226基因的靶向关系。结果与HaCaT细胞(1.089±0.197)相比,A431细胞的miR-378a水平升高至4.435±0.807(P<0.05)。与空白转染组的1.031±0.173相比,增强转染组的miR-378a水平升高至11.846±2.179,而抑制转染组则降低至0.162±0.040;与空白转染组相比,增强转染组转染48、72 h后的增殖活力升高,而抑制转染组则降低;与空白转染组的(7.464±1.281)%相比,增强转染组的凋亡率降低至(3.799±1.279)%,而抑制转染组则升高至(15.435±2.112)%;细胞侵袭实验中,与空白转染组的(62.704±5.639)个相比,增强转染组的穿膜细胞数升高至(89.401±6.358)个,而抑制转染组则降低至(17.346±2.954)个;细胞迁移实验中,与空白转染组的(84.227±5.478)个相比,增强转染组的穿膜细胞数升高至(102.139±7.930)个,而抑制转染组则降低至(54.448±13.068)个;与空白转染组的0.428±0.033相比,增强转染组的CD226水平降低至0.173±0.056,而抑制转染组则升高至0.695±0.020。以上组间差异均有统计学意义(P<0.05)。miR-378a mimics和携带CD226野生型3’端非翻译区(3’UTR)质粒共转染时,荧光素酶活性显著下降(P<0.05);而当miR-378a mimics与携带CD226突变型3’UTR质粒共转染时,荧光素酶活性无显著变化(P>0.05)。结论CSCC细胞中miR-378a高表达并发挥促癌作用,下调该miRNA水平可抑制CSCC细胞增殖、迁移和侵袭能力并诱导凋亡,可能通过靶向CD226来发挥促癌作用,在CSCC靶向治疗中有一定的应用前景。     

微小RNA-92a靶向PTEN／Akt信号通路对鼻咽癌细胞增殖与凋亡的影响

目的 探讨微小RNA-92a（miR-92a）通过靶向PTEN／Akt信号通路对鼻咽癌HONE1细胞增殖与凋亡的调控作用。方法 采用Lipofectamine脂质体法向HONE1细胞转染miR-92a抑制剂（miR-92a组）和阴性对照（NC组）,设不行转染的HONE1细胞为对照组。转染成功后用实时荧光定量PCR（QPCR）检测miR-92a在不同细胞组中的表达情况;MTT法检测各组细胞的增殖率;流式细胞技术检测各组细胞的凋亡率;Western blotting实验检测PTEN／Akt信号通路中PTEN和p-Akt蛋白的表达量。结果 转染48 h miR-92a组与对照组和NC组相比,miR-92a的表达量明显降低（P＜0.05）,而对照组和NC组的差异无统计学意义（P＞0.05）。miR-92a组转染24、48、72、96 h的细胞增殖率分别为（84.51±2.74）％、（77.21±3.55）％、（62.07±3.57）％和（49.25±4.15）％,其中72、96 h的细胞增殖率低于对照组和NC组（P＜0.05）;miR-92a组转染48 h的细胞凋亡率为（46.12±1.79）％,高于对照组的（6.99±0.72）％和NC组的（8.42±0.81）％,差异有统计学意义（P＜0.05）;miR-92a组转染48 h PTEN和p-Akt蛋白的表达水平分别为0.61±0.12和0.37±0.09,对照组分别为0.41±0.11和0.73±0.14,NC组分别为0.39±0.08和0. 68±0.07,与对照组和NC相比,miR-92a组细胞的PTEN表达升高,p-Akt表达降低,差异有统计学意义（P＜0.05）。结论 通过抑制miR-92a的表达来抑制鼻咽癌细胞增殖且促进其凋亡,可能是通过负调控PTEN／Akt信号通路来实现的。     

微小RNA-320在肿瘤中的研究进展

微小RNA（miRNAs）是一类细胞内源性表达的小分子非编码RNA,具有转录后调控基因表达的作用。近年来,在结肠癌、乳腺癌和胆管癌等多种肿瘤中发现miR-320表达下调,提示miR-320可通过抑制细胞增殖和侵袭等方式抑制肿瘤进展。对于miR-320调控的靶基因及相关信号通路的深入研究,有助于阐明肿瘤发生发展的分子机制,并有望为临床肿瘤治疗提供新的靶点。本文就miR-320在肿瘤中的研究进展作一综述。     

MircoRNA-26a与肿瘤

微小RNA(microRNAs,miRNAs)是一类由20个-22个核苷酸组成的小片段非编码RNA,通过靶向结合基因mRNA的3'非翻译区(3'-UTR)调控其表达.许多研究报道miRNAs参与肿瘤的发生发展.MiR-26a在不同的肿瘤中发挥不同的作用,在肿瘤增殖、转移侵袭、血管形成、生物代谢及诊断预后中都有作用.本文就miR-26a与肿瘤关系的研究进展进行综述.     

微小RNA-203与肿瘤

微小RNA (miR)-203具有细胞干性抑制潜能,可通过抑制细胞干性相关转录因子的表达来调节细胞上皮样分化.研究表明miR-203在多种肿瘤组织中表达异常,包括膀胱癌、乳腺癌、结肠癌和胰腺癌等.miR-203通过调节细胞增殖、分化和凋亡在肿瘤的发生发展中发挥重要作用.     

微小RNA-30c在肾癌组织中的表达及其临床意义

目的   探讨微小RNA-30c（miR-30c）在肾癌组织中的表达及其临床病理特征和预后关系。方法   收集本院2012年1月至2015年12月经手术切除的95例肾癌组织和82例癌旁组织标本,采用实时定量PCR（QPCR）检测以上组织中miR-30c水平,分析miR-30c表达与肾癌临床病理参数（性别、年龄、TNM分期、淋巴结转移、远处转移和T分期）及预后的关系,采用Cox回归模型进行多因素分析。结果  QPCR检测发现95例肾癌组织中miR-30c的表达水平为0.561±0.378,低于82例癌旁组织的1.220±0.820（P＜0.05）。肾癌组织中miR-30表达与性别、远处转移和年龄均无关（P＞0.05）,而与TNM分期、淋巴结转移和T分期均有关（P＜0.05）,其中Ⅲ～Ⅳ期、淋巴结转移及T3～T4期的miR-30c相对表达量分别为0.422±0.219、0.442±0.299和0.408±0.194,均低于Ⅰ～Ⅱ期、无淋巴结转移及T1～T2期的0.671±0.439、0.619±0.400和0.654±0.431（P＜0.05）;Ⅲ～Ⅳ期、淋巴结转移及T3～T4期的miR-30c高表达率分别为21.43％（9／42）、16.13％（5／31）和19.44％（7／36）,均低于Ⅰ～Ⅱ期、无淋巴结转移及T1～T2期的41.51％（22／53）、16.13％（26／64）和40.68％（24／59）,差异均有统计学意义（P＜0.05）。miR-30c低表达组的中位总生存期为37.0个月,短于高表达组（＞60.0个月）,差异有统计学意义（P＜0.05）。Cox回归模型分析发现,TNM分期、淋巴结转移、远处转移、T分期及miR-30c表达均是影响预后的独立因素。结论   肾癌组织中miR-30c低表达,miR-30c参与肾癌的发生发展且与预后有关,其中miR-30c低表达者的预后较差,是影响预后的独立因素。     

微小RNA-126和微小RNA-7在食管鳞状细胞癌中的表达及临床意义

目的探讨微小RNA-126（miR-126）和微小RNA-7（miR-7）在食管鳞状细胞癌（ESCC）中的表达情况及其与ESCC临床病理特征、预后之间的关系。方法采用实时荧光定量聚合酶链反应法检测116例ESCC组织和癌旁正常组织中miR-126和miR-7的表达差异,并对miR-126和miR-7表达情况与ESCC患者临床病理特征、预后的相关性进行统计学分析。结果miR-126不同程度低表达病例数为73（62．9％）,正常表达病例数为35（30．2％）,高表达病例数为8（6．9％）;miR-7不同程度低表达病例数为52（44．8％）,正常表达病例数为35（30．2％）,高表达病例数为29（25．0％）。miR-126和miR-7表达降低组无瘤生存期均较非降低组短（r=4．268,P〈0．05;x2=4．993,P〈0．05）;miR-126低表达与ESCC肿瘤位置、家族史和饮酒相关（Xz=14．564,P〈0．05,x2=5．691,P〈0．05;X2=4．971,P〈0．05）,与性别、年龄、分化程度、浸润程度、淋巴转移、吸烟不相关（均P〉0．05）;miR-7低表达与ESCC的临床病理特征均不相关（均P〉O．05）。结论miR-126和miR-7的低表达可能与ESCC患者预后相关,具有一定的临床检测意义。     

微小RNA-181a在人食管癌EC9706细胞中对靶基因PRDM1调控作用的研究

目的:构建微小RNA(microRNA,miR)-181a的真核表达载体和靶基因PRDM1(encoding PR domain containing1,with ZNF domain)的3’非翻译区(3’-untranslated region,3’UTR)荧光素酶报告载体,在人食管癌EC9706细胞中验证miR-181a对靶基因PRDM1的调控作用。方法:根据miR-181a成熟序列在基因组中的位置及其上下游200个碱基序列,以人食管癌KYSE150细胞基因组DNA为模板设计引物,PCR扩增包含miR-181a前体的序列,克隆到线性化的pMD18-T Simple载体中,经BamHⅠ和EcoRⅠ酶切后亚克隆到质粒pCDNA3.1(+)上,对重组质粒进行酶切鉴定和测序分析。通过实时定量PCR法检测人食管癌EC9706细胞转染重组表达载体pCDNA3.1-miR-181a后成熟miR-181a的表达水平。应用生物信息学预测软件对miR-181a的靶基因进行预测,将候选靶基因PRDM1的3’UTR融合到pMIR荧光素酶基因下游,通过双荧光素酶报告基因检测miR-181a对靶基因PRDM13’UTR的调控作用。将pCDNA3.1-miR-181a表达质粒转染到人食管癌EC9706细胞中,蛋白质印迹法检测其对PRDM1蛋白表达的影响。结果:成功构建miR-181a真核表达载体,实时定量PCR验证表明pCDNA3.1-miR-181a在EC9706细胞中能够过表达成熟miR-181a。生物信息学预测PRDM1可能是miR-181a的靶基因之一,于是构建了PRDM1的3’UTR荧光素酶报告载体pMIR-PRDM1,在此基础上对PRDM1的3’UTR"种子区"进行定点突变。双荧光素酶报告基因分析表明,miR-181a能够作用于PRDM1基因的3’UTR。蛋白质印迹法进一步证实,miR-181a能够抑制性调控内源性PRDM1蛋白的表达。结论:MiR-181a作用于PRDM1基因的3’UTR,在转录后水平上调PRDM1蛋白的表达,提示PRDM1是miR-181a直接调控的靶基因。     

微小RNA-373靶向调控LATS2的表达及其对肝癌细胞HepG2增殖与凋亡的影响

目的 探讨微小RNA-373（miR-373）靶向调控大肿瘤抑制因子2（LATS2）的表达及其对肝癌细胞HepG2增殖与凋亡的影响。方法 采用实时荧光定量PCR（QPCR）法检测HepG2细胞及正常肝细胞LO2中miR-373的表达水平,采用Lipofectamine脂质体法将miR-373抑制剂及阴性对照转染至HepG2细胞并分为抑制剂组和对照组,采用QPCR法检测两组的miR-373水平以评价转染效果,MTT法和流式细胞术分别检测各组细胞的增殖及凋亡情况,Western blotting检测各组凋亡相关基因（Bax和caspase-3）及LATS2的表达情况,采用双荧光素酶报告实验验证miR-373与LATS2间的靶标关系。结果 与LO2细胞相比,HepG2细胞中miR-373的表达水平升高,差异有统计学意义（P＜0.05）;转染后抑制剂组的miR-373水平低于对照组（P＜0.05）,提示抑制HepG2细胞miR 373表达成功;与对照组比较,抑制组的细胞增殖率降低,凋亡率及Bax、caspase-3和LATS2蛋白水平均升高,差异有统计学意义（P＜0.05）;双荧光素酶报告基因实验表明miR-373可显著抑制野生型LATS2-3’UTR质粒转染细胞的荧光素酶活性,而对突变型LATS2-3’UTR质粒转染细胞的荧光素酶活性并无影响。结论 miR-373可靶向调控LATS2的表达,且抑制miR-373表达可抑制HepG2细胞增殖并诱导凋亡,为肝癌的靶向治疗提供一定参考。     

miR-100与肿瘤关系的研究进展

miR-100既可表现出致癌基因的作用,又可表现出抑癌基因的作用,其不仅在多种肿瘤（宫颈癌、前列腺癌、膀胱癌和肺癌等）的发生、发展中发挥重要作用,而且还可增加肿瘤细胞对化疗药物的敏感性。miR-100有望成为肿瘤治疗的新靶点及预测预后的新分子标记。本文结合国内外最新报道对miR-100与肿瘤相关性的研究进展作一综述。     

Stathmin蛋白:一个潜在的肿瘤标志物

人stathmin也称原癌基因蛋白18(oncogene protein 18, Op18), 是一种广泛存在于细胞质的蛋白质,其相对分子质量约为19×10~3,可与微管蛋白结合,参与微管和纺锤体的组装;与细胞的增殖、分化、再生和运动均有关,并具有信号活性调节的功能.近年来有研究发现,stathmin在多种肿瘤中高表达,并可通过调节微管的解聚,促进肿瘤细胞的运动及侵袭.Stathmin翻译后修饰状态的改变,可影响与p53蛋白的相互作用,参与肿瘤的发生发展.目前单独或者联合化疗药物使用的抗stathmin效应剂已用于某些肿瘤的治疗.虽然stathmin与肿瘤病因学的内在联系尚不十分清楚,但其作为一个潜在的肿瘤标志物或药物靶点值得进一步研究探讨.     

Comprehensive bioinformatics analyses identified Homeobox B9 as a potential prognostic biomarker and therapeutic target for gastric cancer

BackgroundThe Homeobox B (HOXB) family promotes tumor progression, but the mechanism of its action in gastric cancer (GC) is unclear. We sought to identify the HOXB family members that are critical to the prognosis of GC patients.MethodsThe Oncomine, Gene Expression Profiling Interactive Analysis (GEPIA), cBioPortal, UALCAN, Kaplan-Meier plotter, and the GeneMANIA databases were used to analyze the messenger RNA (mRNA) expression levels, prognostic value, and gene-gene interaction network of the HOXB9 family members in GC. The expression of HOXB9 in GC and its relationship with various clinicopathological parameters and the prognosis of patients were verified by immunohistochemistry.ResultsThe expression of HOXB3, HOXB5, HOXB6, HOXB7, HOXB9, and HOXB13 mRNA was significantly upregulated in GC. There was a significant correlation between the upregulation of HOXB3, HOXB5, and HOXB9 mRNA and a low overall survival (OS) rate. The high expression of HOXB7, HOXB9, and HOXB13 mRNA was closely correlated to tumor grade and stage. HOXB9 was the HOXB family member most closely related to the occurrence and development of GC. A further analysis showed that HOXB9 might be involved in deoxyribonucleic acid repair and division regulation. A validation study showed that the advanced cancer group had a higher level of HOXB9 expression than the early cancer group. The high expression of HOXB9 in gastric tissue plays an important role in the survival and prognosis of GC patients.ConclusionsHOXB family members have different degrees of abnormal expression in GC. High HOXB9 expression in GC tissues was significantly correlated with a worse prognosis. Thus, HOXB9 is a potential novel biomarker and therapeutic target for GC.     

Kinesin family member 11 is a potential therapeutic target and is suppressed by microRNA-30a in breast cancer

Kinesin family member 11 (KIF11) is a plus end-directed kinesin indispensable for the formation of the bipolar spindle in metaphase, where it objects to the action of minus end-directed molecular motors. Here, we hypothesize that KIF11 might be a therapeutic target of breast cancer and regulated by miR-30a. Cell Counting Kit 8 assays were used to investigate cell proliferation. Invasion assays were used to survey the motility of cells. Kaplan-Meier and Cox proportional analyses were employed for this outcome study. The prognostic significance and performance of KIF11 were validated on 17 worldwide independent microarray datasets and two The Cancer Genome Atlas-Breast Invasive Carcinoma sets. microRNA was predicted targeting KIF11 through sequence alignment in microRNA.org and confirmed by coexpression analysis in human breast cancer samples. Dual-luciferase reporter assays were employed to validate the interaction between miR-30a and KIF11 further. Higher KIF11 mRNA levels and lower miR-30a were significantly associated with poor survival of breast cancer patients. Inhibition of KIF11 by small-hairpin RNA significantly reduced the proliferation and invasion capabilities of the breast cancer cells. Meanwhile, downregulation of KIF11 could enhance the cytotoxicity of adriamycin in breast cancer cell lines MCF-7 and MDA-MB-231. A population study also validated that chemotherapy and radiotherapy significantly improved survival in early-stage breast cancer patients with low KIF11 expression levels. Further bioinformatics analysis demonstrated that miR-30a could interact with KIF11 and validated by dual-luciferase reporter assays. Therefore, KIF11 is a potential therapeutic target of breast cancer. miR-30a could specifically interact with KIF11 and suppress its expression in breast cancer.     

MicroRNA-135在肿瘤中的研究进展

MircroRNA（miRNAs）通过与信使RNA（mRNA）的序列互补来降解靶mRNA并抑制蛋白质翻译,在细胞增殖、分化、发育和凋亡等方面起着重要的作用。许多miRNA可作为原癌基因或抑癌基因,在肿瘤的发生和发展中发挥重要作用。越来越多的研究表明,微miR-135（miR-135）与肿瘤的发生发展密切相关,本文主要对近年来miR-135参与调节肿瘤细胞增殖、凋亡、侵袭、转移、耐药和预后等方面的机制作一综述,期望能对肿瘤的早期诊断、治疗和预后判断有所帮助。     

CD25: A potential tumor therapeutic target

CD25 is the alpha-chain of the heterotrimer IL-2 receptor. CD25 is expressed on the surface of both immune and non-immune cells with different frequencies. For cancers, CD25 is expressed at high levels in many types of hematological malignancies, but at low levels in most solid tumors. CD25 is also highly expressed in activated circulating immune cells and regulatory T cells (Tregs). Infiltration of Tregs in the tumor microenvironment can lead to an imbalanced ratio of effector T cells (Teffs) and Tregs, which is associated with the progression of cancers. A rescued Teff/Treg cell ratio indicates an efficient anti-tumor response to immunotherapy. CD25 as a potential target for the depletion of Tregs is critical in developing new immunotherapeutic strategies. Few articles have summarized the relationships between CD25 and tumors, or the recent progress of drugs targeting CD25. In this paper, we will discuss the structures of IL-2 and IL-2R, the biological function of CD25 and its important role in tumor therapy. In addition, the latest research on drugs targeting CD25 has been summarized, providing guidance for future drug development.     

MicroRNA-30a-3p inhibits tumor proliferation,invasiveness and metastasis and is downregulated in hepatocellular carcinoma

Background

MicroRNAs (miRNAs) are small non-coding RNAs that regulate physiological and pathological processes by suppressing target gene expression. Altered expression of miR-30a-3p has been demonstrated in several cancers. However, little about how miR-30a-3p functions in these cancers has been reported, and the role of miR-30a-3p in hepatocellular carcinoma (HCC) is unknown. The purpose of this study was to identify the role and underlying molecular mechanism of action of miR-30a-3p in HCC.

Methods

A total of 110 HCC patients, primarily treated by surgical removal of tumors, were involved in the study. HCC cell line Bel-7402 was selected to characterize the function of miR-30a-3p in vitro.

Results

Our results showed that in 83.6% of the 110 HCC patients, expression of miR-30a-3p was significantly downregulated (P < 0.0001) in tumors compared to adjacent normal tissues. In a clinicopathological correlation analysis, downregulation of miR-30a-3p correlated with a significantly higher incidence of portal vein tumor thrombus (PVTT, P = 0.009). Moreover, miR-30a-3p markedly inhibited the invasion and migration of Bel-7402 HCC cells in vitro. Furthermore, miR-30a-3p overexpression had an inhibitory effect on cell proliferation, induced apoptosis and increased arrest of cells in the S phase. We further demonstrated that miR-30a-3p regulates HCC cell function by a mechanism involving reduced vimentin and MMP3 expression and restoration of E-cadherin expression.

Conclusions

our data suggest that miR-30a-3p is downregulated in HCC and acts as a tumor suppressor in vitro. Regulation of vimentin, E-cadherin and MMP3 by miR-30a-3p suggests a useful therapeutic strategy for tumors with reduced miR-30a-3p expression.     

Go-Ichi-Ni-San 2: A potential biomarker and therapeutic target in human cancers

Cancer incidence and mortality are increasing globally, leading to its rising status as a leading cause of death. The Go-Ichi-Ni-San (GINS) complex plays a crucial role in DNA replication and the cell cycle. The GINS complex consists of four subunits encoded by the GINS1, GINS2, GINS3, and GINS4 genes. Recent findings have shown that GINS2 expression is upregulated in many diseases, particularly tumors. For example, increased GINS2 expression has been found in cervical cancer, gastric adenocarcinoma, glioma, non-small cell lung cancer, and pancreatic cancer. It correlates with the clinicopathological characteristics of the tumors. In addition, high GINS2 expression plays a pro-carcinogenic role in tumor development by promoting tumor cell proliferation and migration, inhibiting tumor cell apoptosis, and blocking the cell cycle. This review describes the upregulation of GINS2 expression in most human tumors and the pathway of GINS2 in tumor development. GINS2 may serve as a new marker for tumor diagnosis and a new biological target for therapy.     

MicroRNA-143 functions as a tumor suppressor in human bladder cancer T24 cells

MicroRNA (miR)-143 and -145 were down-regulated in human bladder cancer T24 cells. The enforced expression of miR-143 induced growth-suppression in T24 cells through down-regulation of ERK5 and Akt expression at translational level, and chemically-modified synthetic miR-143 (miR-143/BP) exhibited a greater growth inhibitory effect than wild-type miR-143. In addition, the synthetic miR-143/BP induced apoptotic cell death in some of the transfected cells. Furthermore, co-treatment with the synthetic miR-143/BP and cisplatin showed the additive growth-suppressing effect on T24 cells. These findings suggest that the chemically-modified synthetic miR-143 functions as a tumor suppressor in T24 cells by targeting ERK5 and/or Akt.