上调SW480结肠癌细胞miR-513b水平抑制HMGB3表达引起癌细胞增殖抑制并促进其凋亡

正结直肠癌为消化系统常见恶性肿瘤之一,致死率高[1-2]。因此,早期诊断可降低其死亡率。分子标志物不仅仅能提供诊断的依据,而且也可能为临床治疗提供治疗的靶点。高迁移率族蛋白3(high mobility group-box 3,HMGB3)属于高迁移率族蛋白家族,在DNA复制、转录等方面起重要作用,是白血病的致病基因[3],是通过调节细胞周期参与肿瘤发生发展的标志物[4]。微小RNA(microRNA,miRNA)     

黄芩苷通过上调miR-485抑制卵巢癌细胞增殖和侵袭

目的 探讨黄芩苷对卵巢癌细胞增殖和侵袭的影响及机制。方法 将卵巢癌CAOV-3细胞分为对照组、黄芩苷组、黄芩苷+miR-NC组和黄芩苷+miR-485抑制剂组,qRT-PCR检测各组细胞中miR-485的表达,MTT法检测细胞增殖能力,Transwell实验检测细胞侵袭能力,Western blot检测MUC1蛋白表达。应用生物信息学和双荧光素酶报告基因实验验证miR-485和MUC1的靶向关系。结果 与对照组相比,黄芩苷组CAOV-3细胞增殖活性和侵袭能力下降,细胞中miR-485表达升高,MUC1蛋白表达水平降低(P<0.05);与黄芩苷+miR-NC组相比,黄芩苷+miR-485抑制剂组CAOV-3细胞增殖活性和侵袭能力升高,细胞中MUC1蛋白表达升高(P<0.05);双荧光素酶报告基因实验证实miR-485靶向负调控MUC1的表达。结论 黄芩苷能够抑制卵巢癌CAOV-3细胞增殖和侵袭,机制可能与miR-485/MUC1通路有关。     

lncRNA MALAT1通过上调miR-30a-5p促进乳腺癌细胞增殖

目的 探讨lncRNA MALAT1靶向miR-30a-5p对乳腺癌细胞增殖的影响。方法 CCK-8实验检测lncRNA MALAT1对乳腺癌细胞MCF7增殖的影响。建立MCF7荷瘤小鼠模型检测lncRNA MALAT1对荷瘤小鼠肿瘤生长的影响。通过生物信息学网站预测miR-30a-5p是否与lncRNA MALAT1结合,并通过双荧光素酶报告基因实验进行验证。Real time-qPCR检测lncRNA MALAT1过表达对miR-30a-5p表达的影响。CCK-8实验检测lncRNA MALAT1过表达载体和miR-30a-5pmimics共转染到MCF7细胞对细胞增殖的影响。结果过表达lncRNA MALAT1促进MCF7细胞的体外增殖及MCF7细胞荷瘤小鼠肿瘤生长。lncRNA MALAT1与miR-30a-5p结合,且过表达lncRNA MALAT1下调miR-30a-5p的表达。结论 lncRNA MALAT1通过下调miR-30a-5p表达促进乳腺癌细胞增殖。     

黄芩苷通过上调miR-126基因抑制乳腺癌细胞增殖北大核心CSCD

目的:探讨黄芩苷调节miR-126基因对乳腺癌细胞增殖的影响。方法:在乳腺癌细胞MCF-7内进行不同浓度黄芩苷与miR-126 mimic处理,使用CCK-8、MTT、Transwell与流式细胞术(FCM)分别检测细胞活性、生存率、侵袭与转移、凋亡率;免疫荧光技术(IF)、qRT-PCR和Western blot检测细胞中VEGF与TGF-β表达。结果:CCK-8、qRT-PCR、Western blot和IF检测结果表明,本实验中黄芩苷的适宜浓度为50μg/ml。过表达miR-126可显著影响MCF-7细胞的活性、迁移、侵袭和凋亡率。同时,与黄芩苷+NC组和miR-126 mimic组相比,黄芩苷+miR-126组MCF-7细胞活性显著降低。结论:黄芩苷可抑制乳腺癌细胞的增殖,可能与黄芩苷上调miR-126基因有关。     

miR-143下调肝癌细胞TLR2的表达并抑制肝癌细胞增殖和侵袭

目的探讨肝癌组织中小RNA-143(miR-143)的表达,并通过瞬时转染肝癌细胞观察对癌细胞生物学行为的影响。方法收集肝癌组织标本及其对应癌旁组织,实时定量PCR检测miR-143在肝癌组织与对应癌旁组织中的表达情况;人工合成寡义核苷酸miR-143 mimics,体外瞬时转染HepG2肝癌细胞,MTT法检测细胞增殖变化,annexinⅤ-FITC/PI染色结合流式细胞仪检测细胞凋亡变化,TranswellTM侵袭实验检测细胞侵袭力变化,Western blot法检测Toll样受体2(TLR2)、核因子κB(NF-κB)、基质金属蛋白酶2(MMP-2)及MMP-9蛋白表达情况。结果 miR-143在肝癌组织中低表达,通过转染miR-143mimics上调miR-143水平后,可以抑制肝癌细胞增殖、促进肝癌细胞凋亡、抑制肝癌细胞侵袭,并下调TLR2、NF-κB、MMP-2及MMP-9表达。结论 miR-143在肝癌中低表达,肝癌细胞过表达miR-143可下调TLR2、NF-κB、MMP-2及MMP-9表达。     

miR-1271-5p促进大鼠心肌细胞增殖

目的研究miR-1271-5p对大鼠心肌细胞增殖和凋亡的作用,以及对肌浆网钙ATP酶(SERCA2a)表达的影响,为临床上心衰药物研发提供临床前期数据。方法转染miR-1271-5p mimics(模拟物)至大鼠心肌细胞,MTS法检测心肌增殖情况;用实时荧光定量PCR方法检测miR-1271-5p对心肌细胞SERCA2a表达的影响。结果miR-1271-5p转染组心肌细胞活力明显高于对照miRNA转染组(P0.05),但SERCA2a表达水平没有显著差异。结论miR-1271-5p促进心肌细胞增殖,对临床心衰治疗有应用前景。     

miR-155在中介素促进肝癌细胞增殖中的作用

目的 MicroRNAs(miRNAs)在恶性肿瘤发生发展过程中起到重要作用,小分子多肽中介素(Intermedin,IMD)可促进肝癌细胞的增殖活性。本文探讨了miR-155在中介素促进肝癌细胞增殖中的作用。方法以CCK-8检测试剂盒检测SMMC7721肝癌细胞增殖,实时定量PCR方法检测增殖细胞核抗原PCNA和miR-155的表达。结果与中介素可以呈时间和剂量依赖关系诱导肝癌SMMC7721细胞的增殖,同时可显著上调miR-155mRNA表达水平;而阻断miR-155可在一定程度上抑制由中介素诱导的SMMC7721细胞增殖。结论中介素促肝癌细胞增殖作用可能与上调miR-155表达相关。     

干扰LINC00313可促进miR-302的表达影响宫颈癌细胞增殖及凋亡

目的 探讨lncRNALINC00313对宫颈癌细胞增殖、凋亡的影响及其对miR-302的调控作用。方法 采用qRT-PCR法检测宫颈癌组织、癌旁组织中LINC00313的表达水平;体外培养人宫颈癌细胞SiHa,分为si-NC组（转染si-NC）、si-LINC00313组（转染si-LINC00313）、si-LINC00313+anti-miR-302组（共转染si-LINC00313、anti-miR-302）、si-LINC00313+anti-miR-NC组（共转染si-LINC00313、anti-miR-NC）。qRT-PCR检测LINC00313、miR-302表达水平;MTT实验检测细胞增殖;流式细胞仪检测细胞周期及凋亡率;双荧光素酶报告实验检测LINC00313、miR-302的靶向关系;Western blot法检测Bax、Bcl-2蛋白水平。结果 与癌旁组织比较,宫颈癌组织LINC00313的表达水平显著升高（P<0.05）;转染si-LINC00313可明显降低OD值、S期细胞比例及Bcl-2蛋白水平（P<0.05）,提高G0-G1期细胞比例、凋亡...     

Nrf2在食管鳞癌组织中的表达及意义

目的:探讨Nrf2(Nuclear factor E2 p45-related factor2)在食管鳞癌组织中的表达及其与临床病理学特征的关系。方法:采用免疫组化SP法检测Nrf2在32例食管鳞癌,30例癌旁组织,21个阳性淋巴结和24个阴性淋巴结组织中的表达。结果:Nrf2阳性表达主要定位于细胞核中,在食管鳞癌中的阳性表达率为78.13%,显著高于癌旁组织(13.33%),淋巴结癌转移阳性组织中的表达率(66.67%)也显著高于淋巴结癌转移阴性组织中的表达水平(20.83%),均具有统计学意义(P<0.05)。Nrf2的阳性表达随淋巴结的转移度的增加而表达增加(P<0.05),但在不同年龄、性别、TNM分期、肿瘤分化程度及不同部位之间差异无统计学意义。结论:Nrf2在食管鳞癌中高表达,表达的高低与淋巴结转移与否及转移度有关。     

食管鳞癌组织中miRNAs差异表达谱的筛选及研究

目的 筛选并鉴定在食管鳞癌（ESCC）及癌旁组织中miRNAs的差异表达,为进一步阐明其在ESCC发病机制中的作用奠定基础。方法 选取在邯郸市第一医院行食管癌切除术患者新鲜的鳞癌组织及癌旁正常组织,各3例,抽提总RNA,利用miRNAs芯片筛选其中差异表达的miRNAs,并对miR-106b-3p通过进一步RT-qPCR 技术进行验证。结果 miRNAs芯片从配对组织中共筛检出62个差异表达的miRNAs,其中41个miRNAs表达上调,21个miRNAs表达下调,鳞癌组织与癌旁正常组织中差异表达的miRNAs比较,差异有统计学意义（P＜0.05）。miR-106b-3p在ESCC组织中的表达高于癌旁组织,差异有统计学意义（P＜0.05）,与芯片结果一致。结论 食管鳞癌组织中miRNAs的差异表达为进一步研究miRNAs在ESCC发病中的作用奠定了基础;miR-106b-3p的高表达可能与ESCC的发生、发展有关。     

p63蛋白在食管磷状细胞癌中的表达及意义

目的检测p63蛋白在食管癌组织中的表达情况,并探讨其与食管癌临床病理特征的关系。方法采用免疫组化检测p63蛋白在53例食管磷癌及癌旁组织中的表达情况。结果p63蛋白在食管癌组织中的阳性表达率为71.7%（38/53）,而在癌旁食管组织中的阳性表达率为35.8%（19/53）,两者比较有显著性差异（P〈0.01）。p63蛋白的表达与食管磷癌分化程度密切相关（P〈0.05）,其在低分化磷癌中的阳性表达率（89.5%）显著高于在高分化磷癌中的阳性表达率（46.2%）。p63蛋白的表达与食管磷癌的TNM分期、淋巴结转移、浸润深度均无明显相关性（P〉0.05）。结论p63蛋白在食管磷癌组织中呈高表达,表明其可能参与食管磷癌的发生发展。     

The Expression of MicroRNA-155 in Plasma and Tissue Is Matched in Human Laryngeal Squamous Cell Carcinoma

PurposeTumor-associated microRNAs have been detected in cancer, though whether plasma microRNA-155 (miR-155) could be a potential biomarker for laryngeal squamous cell carcinoma (LSCC) prognosis is unclear. We aimed to determine how miR-155 can be used to predict the clinical characteristics of patients with LSCC and correctly diagnose them.ResultsA total of 280 LSCC patients and 560 age- and sex-matched controls were included in the study. The miR-155 level was more up-regulated in LSCC tissue than in the non-tumor tissues (13.6±2.4 vs. 3.1±0.80, p<0.001). Additionally, a significantly higher miR-155 level in plasma samples from LSCC patients than in those of the controls (8.9±1.25 vs. 1.8±0.8, p<0.001) was reported. Tissue miR-155 showed an area under the curve (AUC) of 0.933, with a sensitivity of 82.6% and a specificity of 89.2%. The AUC for plasma miR-155 was 0.757, with a sensitivity of 58.4% and a specificity of 69.5%. When early LSCC in TNM I stage was considered, tissue miR-155 showed an area under the curve of 0.804, with a sensitivity of 85.2% and a specificity of 87.3%.ConclusionThe expression of tissue and plasma miR-155 were significantly up-regulated in patients with LSCC. Our work will serve as a basis for further investigation, preferably large-scale validation in clinical trials.     

Low EphA7 Expression Correlated with Lymph Node Metastasis and Poor Prognosis of Patients with Esophageal Squamous Cell Carcinoma

As a member of the Eph family of receptor tyrosine kinases, EphA7 plays an important role in cancer. However, the expression and significance of Eph receptors in esophageal squamous cell carcinoma (ESCC) remain unclear. Here, we detected the expression of EphA7 by immunohistochemistry in a sample of 352 patients with ESCC, and aimed to investigate the expression status of EphA7 in ESCC and its impact on prognosis. The results showed that low EphA7 expression significantly correlated with lymph node metastases (N0: 29%; N1: 64%. p<0.001), poor degree of tumor differentiation (G1: 31%; G2: 49%; G3: 58%. p=0.009) and pTNM staging (I+II: 33%; III+IV: 58%. p<0.001). Furthermore, in a combined analysis, patients with low EphA7-expressing tumors showed a shorter overall survival than those with high expression, resulting in a five-year overall survival rate of 47.4% vs. 52.6%, respectively (p=0.016). Consequently, patients with a low EphA7 expression have poorer prognosis in ESCC compared with those manifesting high expression.     

Prognostic CpG Methylation Biomarkers Identified by Methylation Array in Esophageal Squamous Cell Carcinoma Patients

Background: Esophageal squamous cell carcinoma (ESCC) is an aggressive cancer with poor prognosis. We aimed to identify a panel of CpG methylation biomarkers for prognosis prediction of ESCC patients.Methods: Illumina''s GoldenGate methylation array, supervised principal components, Kaplan-Meier survival analyses and Cox regression model were conducted on dissected tumor tissues from a training cohort of 40 ESCC patients to identify potential CpG methylation biomarkers. Pyrosequencing quantitative methylation assay were performed to validate prognostic CpG methylation biomarkers in 61 ESCC patients. The correlation between DNA methylation and RNA expression of a validated marker, SOX17, was examined in a validation cohort of 61 ESCC patients.Results: We identified a panel of nine CpG methylation probes located at promoter or exon1 region of eight genes including DDIT3, FES, FLT3, NTRK3, SEPT5, SEPT9, SOX1, and SOX17, for prognosis prediction in ESCC patients. Risk score calculated using the eight-gene panel statistically predicted poor outcome for patients with high risk score. These eight-gene also showed a significantly higher methylation level in tumor tissues than their corresponding normal samples in all patients analyzed. In addition, we also detected an inverse correlation between CpG hypermethylation and the mRNA expression level of SOX17 gene in ESCC patients, indicating that DNA hypermethylation was responsible for decreased expression of SOX17.Conclusions: This study established a proof-of-concept CpG methylation biomarker panel for ESCC prognosis that can be further validated by multiple cohort studies. Functional characterization of the eight prognostic methylation genes in our biomarker panel could help to dissect the mechanism of ESCC tumorigenesis.     

LINC00659靶向miR-149-5p调控食管鳞癌细胞Eca-109的增殖、迁移、侵袭及放射敏感性

目的 探讨长基因间非编码RNA 00659(LINC00659)是否靶向miR-149-5p调控食管鳞癌Eca-109细胞的增殖、 迁移侵袭及放射敏感性.方法 实时定量PCR(RT-qPCR)分析30对食管鳞癌组织、 对照组织中LINC00659和miR-149-5p表达量.将Eca-109细胞分为si-NC组、si-...     

VAV3蛋白在喉鳞状细胞癌中的表达及作用机制

目的 研究VAV3蛋白在喉鳞状细胞癌组织中的表达情况,及其对喉癌临床分型、淋巴转移等影响,分析其可能的机制.方法 利用免疫组织化学方法检查VAV3蛋白在喉鳞状细胞癌组织及喉部非癌组织中的表达,检测VAV3蛋白对喉鳞状细胞癌侵袭,淋巴转移等影响.结果 VAV3蛋白在喉鳞状细胞癌组织中表达为69.76％,在喉部非癌组织中的表达为10％,前者明显高于后者,差异有统计学意义(x2=11.937,P=0.001).它的表达与临床分型、T分型及淋巴结转移有统计学意义(P＜0.05),与性别、年龄、吸烟、分型之间无统计学意义(P＞0.05).结论 VAV3蛋白在喉癌组织中高表达,且可促进喉癌组织侵袭、转移能力.VAV3蛋白高表达,可作为判断喉癌及预后的一种新的分子标志.     

THRB Genetic Polymorphisms Can Predict Severe Myelotoxicity after Definitive Chemoradiotherapy in Patients with Esophageal Squamous Cell Carcinoma

Objective: Chemotherapy-related toxicities are difficult to predict before treatment. In this study, we investigated whether thyroid hormone receptor beta (THRB) genetic polymorphisms can serve as a potential biomarker in patients with esophageal squamous cell carcinoma (ESCC).Methods: Forty-nine Japanese patients with ESCC who received a definitive chemoradiotherapy (CRT) with 5-fluorouracil and cisplatin in conjunction with concurrent irradiation were retrospectively analyzed. Severe acute toxicities, including leukopenia, stomatitis, and cheilitis, were evaluated according to 6 single nucleotide polymorphisms (SNPs) in the gene; the intronic SNPs of rs7635707 G/T, rs6787255 A/C, rs9812034 G/T, and rs9310738 C/T and the SNPs in the 3′-untranslated region (3′-UTR) of rs844107 C/T and rs1349265 G/A.Results: Distribution of the 4 intronic SNPs, but not the 2 SNPs in the 3′-UTR, showed a significant difference between patients with and without severe acute leukopenia. Stomatitis and cheilitis were not associated with any of the 6 analyzed SNPs. Frequency of haplotype of the 4 intronic SNPs reached approximately 97% with the 2 major haplotypes G-A-G-C (73.4%) and T-C-T-T (23.5%).Conclusions: THRB intronic SNPs can provide useful information on CRT-related severe myelotoxicity in patients with ESCC. Future studies will be needed to confirm these findings.