MiRNA-10a is upregulated in NSCLC and may promote cancer by targeting PTEN

MicroRNAs (miRNAs) are involved in human cancer including non-small cell lung cancer (NSCLC). In this study, we compared miRNA expression microarray of SPC-A-1sci (high metastatic) and SPC-A-1 (weakly metastatic) cells. We found that miRNA-10a was up-regulated in NSCLC compared with corresponding normal tissues. High expression of miR-10a was associated with tumor node metastasis and lymph node metastasis. Furthermore, overexpression of miR-10a promoted NSCLC cell proliferation, migration and invasion in vitro. We found that PTEN was a direct target of miR-10a in NSCLC. Also miR-10a activated the PTEN/AKT/ERK pathway. We suggest that miR-10a contributes to NSCLC by targeting PTEN.     

miRNA-130a在肝癌组织中的表达

目的探讨肝癌组织中miRNA-130a的表达及临床意义。方法收集经病理确诊的肝癌进行外科手术切除的28例患者的肝癌组织及癌旁组织,选取16例肝外伤后肝切除患者正常肝组织作为对照,应用实时定量PCR技术检测各组织中miRNA-130a的表达。结果肝癌组织中miRNA-130a表达显著高于癌旁组织及肝外伤切除正常肝组织（P〈0.05）。miRNA-130a表达与肝癌分化程度、临床分期和是否合并肝硬化有关（P〈0.05）。结论 miRNA-130a在肝癌组织中异常高表达,可能参与肝癌的发生发展过程。     

MiRNA-101 inhibits breast cancer growth and metastasis by targeting CX chemokine receptor 7

Whereas miR-101 is involved in the development and progression of breast cancer, the underlying molecular mechanisms remain to be elucidated. Here, we report that miR-101 expression is inversely correlated with the clinical stage, lymph node metastasis and prognosis in breast cancers. Introduction of miR-101 inhibited breast cancer cell proliferation and invasion in vitro and suppressed tumor growth and lung metastasis of in vivo. CX chemokine receptor 7 (CXCR7) is a direct target of miR-101, positively correlating with the histological grade and the incidence of lymph node metastasis in breast cancer patients. The effects of miR-101 were mimicked and counteracted by CXCR7 depletion and overexpression, respectively. STAT3 signaling downstream of CXCR7 is involved in miR-101 regulation of breast cancer cell behaviors. These findings have implications for the potential application of miR-101 in breast cancer treatment.     

MiRNA-1469 promotes lung cancer cells apoptosis through targeting STAT5a

MicroRNAs play key roles in cell growth, differentiation, and apoptosis. In this study, we described the regulation and function of miR-1469 in apoptosis of lung cancer cells (A549 and NCI-H1650). Expression analysis verified that miR-1469 expression significantly increased in apoptotic cells. Overexpression of miR-1469 in lung cancer cells increased cell apoptosis induced by etoposide. Additionally, we identified that Stat5a is a downstream target of miR-1469, which can bind directly to the 3’-untranslated region of the Stat5a, subsequently reducing both the mRNA and protein levels of Stat5a. Finally, co-expression of miR-1469 and Stat5a in A549 and NCI-H1650 cells partially abrogated the effect of miR-1469 on cell apoptosis. Our results show that miR-1469 functions as an apoptosis enhancer to regulate lung cancer apoptosis through targeting Stat5a and may become a critical therapeutic target in lung cancer.     

MiRNA-34a inhibits EGFR-signaling-dependent MMP7 activation in gastric cancer

The molecular mechanism underlying cancer invasiveness and metastasis of gastric carcinoma remains elusive. Here, we reported significant decrease in microRNA (miRNA)-34a and significant increase in phosphorylated epidermal growth factor receptor (EGFR) and matrix metalloproteinase-7 (MMP7) in the resected gastric carcinoma from the patients, compared with adjacent normal tissue. Moreover, strong correlation was detected among these three factors. To examine whether a causal link exists, we used two human gastric carcinoma lines, SNU-5 and HGC27, to study the molecular basis of miRNA-34a, EGFR signaling, and MMP7 activation. We found that EGF-induced EGFR phosphorylation in SNU-5 or HGC27 cells activated MMP7 and consequently cancer invasiveness. Both an inhibitor for EGFR and an inhibitor for Akt significantly inhibited the EGF-induced activation of MMP7, suggesting a phosphatidylinositol 3-kinase (PI3K) signaling cascade dependent pathway. Moreover, miRNA-34a levels were not affected by EGF-induced EGFR phosphorylation. However, overexpression of miRNA-34a antagonized EGF-induced MMP7 activation without affecting EGFR phosphorylation in SNU-5 or HGC27 cells. Taken together, our data suggest that miRNA-34 inhibits EGFR signaling via downstream PI3K signaling cascades to regulate MMP7 expression in gastric carcinoma. Thus, miRNA-34a, EGFR, and MMP7 appear to be promising therapeutic targets for preventing the metastasis of gastric carcinoma.     

miRNA-30b调控ITGB3表达促进乳腺癌细胞凋亡

  目的  探讨微小RNA-30b （microRNA-30b,miRNA-30b）和整合素β3（integrin β3,ITGB3）在乳腺癌组织中的表达及对乳腺癌细胞生物学行为的影响。  方法  选择2016年3月至2019年3月于成都医学院第一附属医院收治的144例包括原位、浸润性和转移性乳腺癌患者的组织标本以及其癌旁组织,同时收集其临床病理资料。采用实时荧光定量PCR（RT-PCR）检测miRNA-30b和ITGB3的表达。构建miRNA-30b高表达的miRNA-30b模拟物和ITGB3高表达的miRNA-30b模拟物-pc DNA3.1-ITGB3,转染乳腺癌MCF-7细胞,同时将转染后的细胞皮下注射至小鼠体内。5-溴脱氧尿嘧啶核苷（BrdU）实验、流式细胞术、Transwell小室侵袭实验检测细胞增殖、凋亡、侵袭,Western blot检测细胞中的ITGB3表达。  结果  RT-PCR结果显示,原位、浸润性和转移性乳腺癌组织中的miRNA-30b相对表达量分别为0.75、0.52和0.23,ITGB3 mRNA分别为2.17、3.76和5.43,与正常组织相比,差异均具有统计学意义（均P<0.001）。miRNA-30b高表达后,BrdU、流式细胞术、Transwell小室侵袭实验、Western blot检测结果显示,癌细胞的增殖、侵袭能力以及ITGB3表达明显降低,凋亡率升高（均P<0.001）。  结论  在乳腺癌组织中miRNA-30b和ITGB3表达异常,miRNA-30b过表达能明显下调ITGB3表达,抑制乳腺癌细胞的增殖、侵袭,并促使其凋亡。      

MiRNA-181c inhibits EGFR-signaling-dependent MMP9 activation via suppressing Akt phosphorylation in glioblastoma

As the most aggressive malignant primary human brain tumor, glioblastoma is noted with extremely poor patient survival. Previous studies have demonstrated that expression of matrix metalloproteinase-9 (MMP9) in glioblastoma cells is critical for cancer metastasis. However, the molecular signaling pathways that control MMP9 activation remain undefined. Here, we reported a strong negative correlation of microRNA (miRNA)-181c levels with either MMP9 levels or activation of epidermal growth factor receptor (EGFR) signaling in glioblastoma patients. EGF-induced activation of EGFR in a human glioblastoma line, A-172 cells, increased MMP9 expression through activation of phosphatidylinositol 3-kinase (PI3K)/Akt pathway, without affecting expression of miRNA-181c. On the other hand, overexpression of miRNA-181c in A-172 cells inhibited MMP9 expression by inhibiting Akt phosphorylation, but not phosphorylation of EGFR receptor. Taken together, these findings suggest that EGFR signaling activates downstream PI3K/Akt to increase MMP9 expression in glioblastoma, while phosphorylation of Akt is a control point by miRNA-181c. Our work thus provides new insights into the molecular basis underlying the metastasis of glioblastoma.     

MiRNA-1297与AEG-1在肿瘤中的研究进展

摘 要：MircoRNA是一种基因表达的调节剂,其表达的改变导致癌基因的激活或肿瘤抑制基因的失活。星形细胞上调基因-1（AEG-1）被认为是一种癌基因,具有调节各种人类癌症中的细胞增殖、转移、转化、血管生成和放化疗抗性等,已经在乳腺癌、肝癌、前列腺等多种癌细胞中发现过表达,且与肿瘤进展相关的PI3K-AKT、NF-κB、MAPK和Wnt多种信号通路中发挥作用。miR-1297是一种具有肿瘤抑制作用的mircoRNA,其抑制或过表达影响与肿瘤增殖、侵袭有关的AEG-1、PTEN、Meg3等基因。     

Targeting Epidermal Growth Factor Receptor by MiRNA-145 Inhibits Cell Growth and Sensitizes NSCLC Cells to Erlotinib

Background: Despite effective activity of epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs), such as erlotinib, all non-small cell lung cancer (NSCLC) patients eventually acquire resistance to these agents. Studies have demonstrated that down-regulation of miRNA-145 leads to enhancement of EGFR expression, cell proliferation and metastasis. The aim of this study was to investigate the effect of miRNA-145 on sensitivity of the A549 NSCLC cells to erlotinib. Methods: Quantitative real-time PCR was used to examine the effect of miRNA-145 on EGFR expression. The effect of miRNA-145 on cell growth and sensitivity the lung cancer cells to erlotinib was examined by trypan blue and MTT assays, respectively. The combination index was calculated using the non-constant method of Chou-Talalay. Apoptosis was determined by ELISA cell death assay. Results: We found that miRNA-145 was markedly suppressed the expression of EGFR and inhibited the cancer cell growth, relative to blank control and negative control miRNA (p<0.05). Pretreatment with miRNA-145 synergistically enhanced the sensitivity of the lung cancer cells to erlotinib. Results of apoptosis assay revealed that miRNA-145 can induce apoptosis and increase the erlotinib-mediated apoptosis. Conclusions: Our data demonstrate that miRNA-145 play a critical role in the lung cancer cell growth, survival and EGFR-TKIs resistance possibly by regulation of EGFR. Therefore, miRNA-145 replacement therapy can become a new therapeutic strategy in lung cancer.     

MiRNA-7 Replacement Effect on Proliferation and Tarceva-Sensitivity in U373-MG Cell Line

Background: Deregulation of the EGFR signaling pathway activity has been shown to can be effective in resistance to EGFR-TKIs, such as Tarceva (erlotinib), in glioblastoma cells. In addition, reports have shown that the reduction of miRNA-7 expression levels is associated with an increase in the expression of EGFR. Here, we evaluated the effect of miRNA-7 on EGFR expression and sensitivity of the U373-MG glioblastoma to erlotinib. Methods: The effect of miRNA-7 on EGFR expression was examined using RT-qPCR and western blotting. Trypan blue and MTT assays were performed to explore the effect of treatments on cell growth and survival, respectively. The combination index analysis was used to evaluate the interaction between drugs. Apoptosis was measured by ELISA cell death assay. Results: We showed that miRNA-7 markedly inhibited the expression of EGFR and decreased the growth of glioblastoma cells, relative to blank control and negative control miRNA (p < 0.05). Introduction of miRNA-7 synergistically increased the sensitivity of the U373-MG cells to erlotinib. Results of apoptosis assay demonstrated that miRNA-7 can trigger apoptosis and enhance the erlotinib-mediated apoptosis. Conclusions: Our results show that miRNA-7 plays a critical role in the growth, survival and sensitivity of the U373-MG cells to erlotinib by targeting EGFR. Thus, miRNA-7 replacement therapy can become an effective therapeutic procedure in glioblastoma.     

Up-Regulation of MiRNA-125a-5p Inhibits Cell Proliferation and Increases EGFR-TKI Induced Apoptosis in Lung Cancer Cells

Background: Despite the dramatic efficacy of erlotinib, an EGFR tyrosine kinase inhibitor (TKI), most of non-small cell lung cancer (NSCLC) patients ultimately acquire resistance to this agent. Different studies indicated that miRNA-125a-5p is down-regulated in human lung cancer cells and may function as a tumor suppressor by targeting EGFR. However, the biological function of miRNA-125a-5p in NSCLC resistance to EGFR-TKIs is not fully understood. In this study the effect of miRNA-125a-5p on cell proliferation, apoptosis and sensitivity of the A549 lung cancer cells to erlotinib was investigated. Methods: After miRNA-125a-5p transfection, the expression levels of EGFR mRNA were measured by QRT-PCR. Trypan blue assays were performed to evaluate the proliferation of the A549 lung cancer cells. The cytotoxic effects of miRNA-125a-5p and erlotinib, alone and in combination, were determined using MTT assay. Combination index study was performed using the method of Chou-Talalay. Apoptosis was assessed using an ELISA cell death assay kit. Results: MiRNA-125a-5p clearly reduced the expression of EGFR mRNA in a time dependent manner, causing marked cell proliferation inhibition and spontaneous apoptosis (p<0.05, relative to control). Pretreatment with miRNA-125a-5p synergistically increased the cytotoxic effect of erlotinib and decreased its IC50. Furthermore, miRNA-125a-5p significantly enhanced the apoptotic effect of erlotinib. Negative control miRNA had no significant effect on biological parameter of the tumor cells. Conclusions: Our data suggest that suppression of EGFR by miRNA-125a-5p can effectively trigger apoptosis and overcome EGFR-TKs resistance of lung cancer cells. Therefore, miRNA-125a-5p may be a potential therapeutic adjuvant in patients with lung cancer.     

肾癌组织中MiRNA-148b的表达及临床意义

目的探讨miRNA-148b在肾癌组织的表达及临床意义。方法选取2012年3月至2014年3月间潍坊医学院附属医院泌尿外科收治的40例肾癌患者,采用实时定量聚合酶链反应(PCR)检测40例患者肾癌及癌旁正常肾组织中miRNA-148b的表达情况,并研究其与临床分期、病理分级之间的关系。结果与配对的癌旁正常肾组织相比,肾癌组织中miRNA-148b的相对表达下降,差异有显著统计学意义(P<0.01)。不同临床分期和病理分级的肾癌组织中miRNA-148b的相对表达差异无统计学意义(P>0.05)。结论 miR-148b在肾癌组织中的表达水平较癌旁正常肾组织明显降低,但与肾癌的病理分级及临床分期无明显相关性。     

MiRNA-155 regulates lymphangiogenesis in natural killer/T-cell lymphoma by targeting BRG1

Background: miR-155 was up-regulated in natural killer/T-cell lymphoma (NKTCL), an aggressive malignancy, and correlated with disease progression. However, minimal is known on biological activities and underlying mechanisms of miR-155 in NKTCL. In this study, we examined BRG1, a potential target of miR-155, and focused on the miR-155/BRG1 signaling in regulating lymphangiogenesis of NKTCL.

Methods: The expression of miR-155, BRG1, VEGFC, and VEGFD was compared between two NKTCL cell lines and normal NK cells. The critical role of miR-155 and STAT3 was assessed using miR-155 inhibitor and STAT3 inhibitor S31-201, respectively. Two biological phenotypes, apoptosis and pro-lymphangiogenesis, were examined in vitro by flow cytometry and lymphatic tube formation, respectively, and in vivo using an NKTCL xenograft model.

Results: The miR-155 level negatively correlated with BRG1, but positively with VEGFC in normal NK as well as two NKTCL cell lines. Targeting miR-155 in NKTCL cells significantly boosted BRG1 expression and decreased the activated STAT3 or VEGFC level, leading to enhanced apoptosis and reduced lymphangiogenesis. STAT3 acted downstream of BRG1 and essentially regulated miR-155-mediated up-regulation of VEGFC and pro-lymphangiogenesis. In vivo, targeting miR-155 inhibited primary xenograft growth as well as tumor-associated lymphangiogenesis.

Conclusions: By inhibiting BRG1 expression, miR-155 activated STAT3/VEGFC signaling and promoted lymphangiogenesis. In addition, miR-155 also controlled the viability of NKTCL cells. Therefore, targeting miR-155 provides a novel therapy for NKTCL.     

MiR-129-5p靶向HMGB1抑制骨肉瘤细胞增殖和迁移

目的 探讨miR-129-5p对骨肉瘤(OS)细胞增殖和迁移的影响以及对HMGB1的调控作用.方法 RT-PCR和Western blot法分别检测骨肉瘤细胞株MG-63、Saos-2和成骨细胞hFOB1.19中miR-129-5p和HMGB1的表达.生物信息学预测miR-129-5p与HMGB1基因是否存在结合位点,...     

miRNA-375靶向调控Notch1基因对胰腺癌细胞增殖、迁移的作用机制研究

摘 要：［目的］ 探讨miRNA-375靶向调控Notch1基因对胰腺癌(PAAD)细胞增殖、迁移的作用机制。［方法］ 选取诊断为PAAD并行胰腺外科手术患者60例,取癌组织和癌旁组织行HE染色、免疫组织化学法和免疫荧光检测,采用PCR法检测胰腺癌组织中miRNA-375和Notch1表达量,使用transwall侵袭实验法和MTT试剂盒检测胰腺癌细胞增殖、迁移和侵袭性,采用生物信息学分析和预测miRNA-375调控胰腺癌细胞Notch1的表达。［结果］ miRNA-375表达水平在癌组织（1.00±0.07）与癌旁组织（1.65±0.14）中有显著性差异（P<0.05）;miRNA-375表达水平与肿瘤大小、TNM分期、淋巴结转移和远处转移有关（P<0.05）。qRT-PCR检测显示PANC-1细胞和BxPC-3细胞中miRNA-375 mimics表达水平显著增加（P<0.05）,转染3天后PANC-1细胞和BxPC-3细胞增殖被显著抑制（P<0.05）,抑制率分别为35.93%和27.71%;转染miRNA-375使其表达水平上升可显著抑制PANC-1细胞和BxPC-3细胞的迁移能力。miRNA-375高表达使PANC-1细胞水解基质胶穿透到小室下面的细胞减少35.35%,BxPC-3细胞减少33.04%（P<0.05）。miRNA-375可以调控Notch1D 表达。利用双荧光素酶报告基因法检测miRNA-375与Notch1 mRNA的作用关系,转入突变3′UTR质粒的细胞中荧光素酶活性显著降低（t=5.633,P=0.039;t=5.347,P=0.031）,miRNA-375直接与Notch1 mRNA的3′UTR相互作用。［结论］胰腺癌细胞中miRNA-375靶向调控Notch1表达。miRNA-375/Notch1通路能够抑制胰腺癌细胞的增殖、成瘤及侵袭转移能力,在胰腺癌细胞中发挥抑癌作用。     

miRNA-21调节PTEN表达影响宫颈癌HeLa细胞的生物学行为

目的： 探讨抑制miRNA-21表达对宫颈癌HeLa细胞中PTEN的表达及细胞增殖、侵袭能力的影响。 方 法： 以脂质体介导anti-miRNA-21（anti-miRNA-21转染组）、anti-miRNA-21-neg（阴性对照组）转染HeLa细胞,同时设空白对照组（未转染组）。应用Real-time PCR技术检测3组细胞中miRNA-21的表达,Western blotting 检测3组细胞中PTEN的表达,MTT法检测3组细胞的增殖能力,Transwell实验检测3组细胞的侵袭能力。结果： Anti-miR-21转染组与阴性对照组相比,HeLa细胞中miRNA-21的表达量明显降低\[（0.187±0.027）vs （0.861±0.144）,P＜0.01\]。转染 anti-miRNA-21 96 h后,HeLa细胞增殖抑制率明显升高\[（49.44±1.97）% vs （4.36±0.64）%,P＜0.01\]。Anti-miR-21转染组与阴性、空白对照相比, Hela细胞的侵袭细胞数明显减少\[（29.4±2.1）vs （40.4±2.9）、（41.2±2.6）个,均P＜0.01\];PTEN蛋白的表达则明显增加\[（1766.00±35.56）vs（726.00±5.48）、（729.25±17.73）,均P<0.01\]。 结论： 抑制miRNA-21的表达后,宫颈癌HeLa细胞增殖、侵袭能力明显下降,其机制可能与上调PTEN的表达有一定关系。     

MiRNA-22在不同分期浆液性卵巢腺癌中的表达以及与化疗药物耐药性关系的研究

摘 要：［目的］ 探讨miRNA-22在不同分期浆液性卵巢腺癌的表达以及与化疗耐药性关系。［方法］ 使用基因芯片技术筛选出差异化表达的miRNA,再用实时荧光定量聚合酶链反应（RT-PCR）检测miRNA-22在人卵巢癌细胞株（SKOV3）、紫杉醇稳定耐药卵巢浆液性囊腺癌细胞亚株（SKOV3-TR30）、人正常卵巢上皮细胞株（IOSE80）及50例确诊为浆液性卵巢腺癌的患者、25例正常人血清中的表达水平,并分析其与患者临床分期的关系以及用药前后的关系。［结果］ miRNA-22在SKOV3-TR30耐药株中的表达明显高于SKOV3敏感株中的表达水平（P<0.05）,IOSE80正常卵巢上皮细胞株中miRNA-22的表达水平明显低于SKOV3敏感株（P<0.05）;在FIGO手术病理不同分期中的表达,Ⅰ、Ⅱ、Ⅲ、Ⅳ期表达水平依次增高;经统一的化疗方案治疗后,化疗无效组的表达水平高于化疗有效组的表达水平（P<0.05）。［结论］ miRNA-22可能是卵巢癌患者潜在的肿瘤标志物,miRNA-22的高表达可能与癌细胞的过度增殖与转移、侵袭有关,具有癌基因与抑癌基因的双侧作用,且可能与化疗药物的耐药性密切相关。