miR-185 Inhibits the Proliferation and Invasion of Non-Small Cell Lung Cancer by Targeting KLF7

MicroRNAs (miRNAs) are short endogenous noncoding RNAs that frequently play vital roles in many cancer types. Herein we demonstrated that miR-185 was remarkably downregulated in NSCLC tissues compared with adjacent normal tissues. A lower level of miR-185 was associated with lymph node metastasis. Functional assays showed that upregulation of miR-185 inhibited the proliferation, colony formation, and invasion capacities of NSCLC cells in vitro. Furthermore, we found that miR-185 suppressed the epithelial–mesenchymal transition (EMT) process. Bioinformatics analysis and luciferase reporter gene assays revealed that Kruppellike factor 7 (KLF7) was the target of miR-185. Overexpression of miR-185 reduced the expression of KLF7 in NSCLC cells. Upregulation of KLF7 partly neutralized the inhibitory effects of miR-185 on the proliferation and invasion of NSCLC. Additionally, we confirmed that miR-185 suppressed the tumor growth of NSCLC A549 cells in vivo. Taken together, these results demonstrate that miR-185 acts as a suppressor by targeting KLF7 in NSCLC.     

miR-449a Suppresses Tumor Growth,Migration, and Invasion in Non-Small Cell Lung Cancer by Targeting a HMGB1-Mediated NF-kB Signaling Pathway

MicroRNAs (miRNAs) have been reported to be involved in many human cancers and tumor progression. The dysregulation of miR-449a is found in many types of malignancies and is associated with tumor growth, migration, and invasion. However, its expression and function in non-small cell lung cancer (NSCLC) still remains unclear. In our study, miR-449a was found to be downregulated in both NSCLC tissues and cell lines, and low miR-449a expression was obviously associated with tumor differentiation, TMN stage, and poor overall survival (OS). Moreover, we demonstrated that miR-449a could inhibit tumor proliferation, migration, and invasion in NSCLC. We also confirmed that HMGB1 was a direct target gene of miR-449a in NSCLC with dual-luciferase reporter assay, and upregulation of HMGB1 could reverse the miR-449a-induced suppression of growth, migration, and invasion in NSCLC cells. Last, we found that miR-449a suppressed tumor initiation and development through the NF- B signaling pathway. These results indicate that miR-449a functions as a tumor suppressor in NSCLC by targeting the HMGB1-mediated NF- B signaling pathway in NSCLC.     

miR-769-5p在非小细胞肺癌中的表达及其对细胞增殖、迁移与侵袭能力的影响

目的 探讨微小RNA-769-5p （miR-769-5p） 在非小细胞肺癌（NSCLC）细胞株中的表达及其对细胞增殖、迁移和侵袭能力的影响。方法 采用实时荧光定量PCR（QPCR）技术检测miR-769-5p 在5种NSCLC细胞（A549、NCI-H1299、NCI-H157、ANIP-973和GLC-82）中的相对表达量,选择相对表达量最低和最高的细胞株分别转染miR-769-5p mimics／miR-769-5p inhibitor,另设miRNA mimics control／inhibitor control对照组;采用MTS 实验、克隆形成实验及Transwell 小室实验检测miR-769-5p对NSCLC 细胞增殖、克隆形成、侵袭和迁移能力的影响。结果 miR-769-5p在A549、NCI-H1299、NCI-H157、ANIP-973 及GLC-82细胞中的相对表达量分别为 0.06、0.40、0.09、1.04和2.31。在miR-769-5p表达水平最低的A549细胞中瞬时转染miR-769-5p mimics,在miR-769-5p表达水平最高的GLC-82细胞中瞬时转染miR-769-5p inhibitor。MTS结果显示,与对照组比较,miR-769-5p mimics 能够抑制A549细胞的增殖 （P＜0.05）,而miR-769-5p inhibitor能够促进GLC-82细胞的增殖 （P＜0.05）。平板克隆实验结果显示,与对照组比较,miR-769-5p mimics 能够抑制A549细胞的平板克隆形成数 （124.7±14.7 vs. 399.4±46.0,P＜0.05）;而miR-769-5p inhibitor能够促进GLC-82细胞的平板克隆形成数 （555.7±29.7 vs. 366.3±28.7,P＜0.05）。Transwell实验结果显示,与对照组迁移和侵袭跨膜细胞数比较,miR-769-5p mimics能够抑制A549细胞的迁移和侵袭 （94.4±18.1 vs. 157.8±22.9,84.4±15.1 vs. 135.6±16.7,P＜0.05）;miR-769-5p inhibitor能够促进GLC-82细胞的迁移和侵袭 （226.7±40.3 vs. 153.3±38.7,196.7±39.1 vs. 138.9±40.4,P＜0.05）。结论 miR-769-5p可以抑制NSCLC细胞的增殖、侵袭和迁移,可能成为NSCLC的潜在靶点。     

非小细胞肺癌血浆miR-223、miR-93和miR-218的表达及其临床意义

目的 探讨非小细胞肺癌（NSCLC）患者血浆miR-223、miR-93和miR-218水平,分析三者与NSCLC患者临床病理学特征的关系。方法 收集本院收治85例NSCLC患者未经治疗前的血浆标本（NSCLC组）,采用实时定量RT-PCR（qRT-PCR）法检测血浆中miR-223、miR-93和miR-218水平,收集NSCLC患者的临床病理学参数,比较三者不同水平的临床病理参数并分析三者间的关系,采用受试者工作特征曲线（ROC）评价血浆miR 223、miR 93和miR 218水平检测在NSCLC诊断中的临床价值。同时选取同期的90例健康体检者的血浆标本作对照（对照组）。结果 NSCLC组的血浆miR-223和miR-93水平高于对照组,miR-218水平低于对照组,差异均有统计学意义（P均＜0.05）;miR-223水平与TNM分期、肿瘤大小有关,miR-93水平与组织学类型、淋巴结转移有关,miR-218水平与肿瘤大小、分化程度有关,以上差异均有统计学意义（P＜0.05）;NSCLC患者血浆中miR-223水平与miR-93呈正相关（r=0.411）,miR-223、miR-93分别与miR-218呈负相关（r=-0.361,r=-0.451）,差异均有统计学意义（P＜0.05）;血浆miR-223、miR-93和miR-218诊断NSCLC的AUC、灵敏度和特异度分别为0.926、95.2％和87.1％,0.912、88.4％和92.5％,0.941、92.7％和84.4％,均高于CEA的0.774、75.1％和66.2％,且miR-223、miR-93和miR-218联合诊断的效能高于单独检测。结论 NSCLC患者血浆中miR-223和miR-93呈高表达,miR-218呈低表达,与临床病理学参数有关,且在NSCLC诊断中有一定的价值,可用于辅助NSCLC的诊断和病情评估。     

VASH2 Promotes Cell Proliferation and Resistance to Doxorubicin in Non-Small Cell Lung Cancer via AKT Signaling

Vasohibin2 (VASH2), a proangiogenic factor, has been demonstrated to play an oncogenic role in some common human cancers. However, the detailed function of VASH2 in non-small cell lung cancer (NSCLC) has not previously been studied. In this study, we found that VASH2 was significantly upregulated in NSCLC tissues and cell lines, and its increased expression was associated with NSCLC progression and poor prognosis of patients. Knockdown of VASH2 markedly inhibited cell proliferation and P-glycoprotein expression in NSCLC cells. Overexpression of VASH2 enhanced cell proliferation, P-glycoprotein expression, as well as doxorubicin resistance in NSCLC cells. Moreover, the expression levels of VASH2 were significantly increased in newly established doxorubicin-resistant NSCLC cells. Molecular mechanism investigation revealed that inhibition of VASH2 expression in NSCLC cells suppressed the activity of AKT signaling, and overexpression of VASH2 enhanced the activity of AKT signaling. We further showed that downregulation of AKT signaling activity using AKT inhibitor LY294002 markedly inhibited NSCLC cell proliferation and resistance to doxorubicin induced by VASH2. In conclusion, the findings in the present study indicate that VASH2 promotes NSCLC cell proliferation and resistance to doxorubicin via modulation of AKT signaling. Thus, we suggest that VASH2 may become a potential therapeutic target for the treatment of NSCLC.     

miR-181c-5p靶向TRIM71调控非小细胞肺癌细胞A549的凋亡研究

目的 探讨miR-181c-5p调控非小细胞肺癌细胞A549的潜在机制.方法 检测非小细胞肺癌细胞A549和人正常肺细胞HLF-a中miR-181c-5p的表达水平.过表达或敲低miR-181c-5p后,检测非小细胞肺癌细胞A549的凋亡水平.通过miRDB在线分析miR-181c-5p潜在底物,过表达miR-181c...     

非小细胞肺癌组织中p63蛋白的表达及其意义

目的 检测p63蛋白在非小细胞肺癌（NSCLC）组织中的表达情况，并探讨其与NSCLC临床病理特征的关系。方法 采用免疫组化（S-P法）检测p63蛋白在76例NSCLC及12例癌旁肺组织中的表达情况。结果 p63蛋白在NSCLC组织中的阳性表达率为55。3％（42／76），而在癌旁肺组织中无阳性表达，两者比较有非常显著性差异（P〈0．01）。p63蛋白的表达与NSCLC病理类型密切相关（P〈0.01），其在肺鳞癌中的阳性表达率（80．0％）显著高于在肺腺癌（33.3％）和肺腺鳞癌（36.4％）中的阳性表达率；在不同分化程度的肺鳞癌中p63的阳性表达率相互比较有显著性差异（P〈0.05）。p63蛋白的表达与NSCLC的TNM分期、淋巴结转移、术后复发及生存期均无明显相关（P〉0.05）。结论 p63蛋白在NSCLC中表达上调，可能是1个有价值的诊断肺鳞癌和判断肺鳞癌分化的标志物。     

Formononetin Inhibits Non-Small Cell Lung Cancer Proliferation via Regulation of mir-27a-3p through p53 Pathway

Objective: The aim of the present study was to investigate the anti-tumor effects of formononetin on human nonsmall cell lung cancer (NSCLC) and its potential molecular mechanism. Methods: A549 cells were treated withdifferent concentrations of formononetin, then detected the cell proliferation, apoptosis and the expression ofHIPK2 respectively by MTT assay, flow cytometry analysis and RT-qPCR. Then the interaction between miR-27a-3p and its target gene HIPK2 was verified through luciferase reporter assay. The expression of miR-27a-3p, HIPK2, and p53 was detected after being treated with different formononetin concentrations by RT-qPCRand western blot. Results: The results showed that formononetin significantly inhibited the proliferation andinduced the apoptosis of A549 cells in a time- and dose-dependent manner. miR-27a-3p targeted HIPK23’UTR and inhibited the expression of HIPK2. Also, formononetin-treated A549 cells were remarked with asignificant decline in the expression of miR-27a-3p, accompanied with growth of HIPK2, and subsequently areduction of p53. Conclusions: The study indicates that miR-27a-3p might pose regulated effects on theHIPK2/p53 pathway, resulting in formononetin’s anti-carcinogenic effects on NSCLC in vitro.     

miR-223-3p通过调控ECT2诱导非小细胞肺癌细胞凋亡的实验研究

目的：探讨miR-223-3p对非小细胞肺癌细胞增殖与凋亡的影响及其可能的作用机制。方法：选取24例非小细胞肺癌患者的癌及癌旁组织,实时定量PCR检测miR-223-3p的相对表达水平,免疫组化检测分析上皮细胞转化序列2（ECT2）蛋白的表达水平,并对两者进行相关性分析。将人非小细胞肺癌A549细胞分为正常对照组、转染对照组、miR-223-3p过表达组,其中正常对照组细胞未作任何处理,转染对照组细胞转染空质粒载体,miR-223-3p过表达组转染miR-223-3p mimics。MTT实验和平板集落形成实验检测各组细胞的增殖率,流式细胞术检测各组细胞的凋亡率,Western blot实验检测ECT2蛋白的表达水平;采用siRNA转染法沉默A549细胞中的ECT2后,检测细胞增殖及凋亡的变化。结果：miR-223-3p在癌组织中表达显著低于癌旁组织,而癌组织中ECT2蛋白表达高于癌旁组织,二者存在负相关关系（r=-0.666,P < 0.05）;与正常对照组和转染对照组细胞相比,miR-223-3p过表达组细胞的增殖率降低（P < 0.05）,ECT2蛋白表达显著降低,而细胞凋亡率显著升高（P < 0.05）;沉默ECT2对细胞增殖和凋亡的影响与过表达miR-223-3p基本一致。结论：miR-223-3p可能通过负向调控ECT2的表达,进而抑制非小细胞肺癌细胞增殖并诱导其凋亡,其作用机制有待进一步研究。     

miR-1266-5p and miR-185-5p Promote Cell Apoptosis in Human Prostate Cancer Cell Lines

Objective: Small non-coding RNA molecules are dysregulated in prostate cancer (PCa). In our previous study,downregulation of miR-1266 and miR-185 was demonstrated in PCa tissues and cell lines. The aim of the presentstudy was to investigate whether miR-1266 and miR-185 are involved in the regulation of B-cell lymphoma (BCL) 2and BCL2L1, respectively, and whether transfection of PCa cell lines with miR-1266 and miR-185 mimics can altertumorigenic phenotypes. Methods: In order to investigate the regulation of BCL2 and BCL2L1 mRNA levels bymiR-1266 and miR-185, respectively, a luciferase reporter assay was used. Real-time PCR was also used to analyzechanges in the levels of BCL2 and BCL2L1 mRNAs in PCa cell lines following transfection with synthetic miR-1266and miR-185. Cell apoptosis was determined by Annexin V protein expression analysis via flow cytometry. In additionto the MTT assay, a cell proliferation assay was performed. Result: A luciferase assay confirmed that the BCL2 andBCL2L1 genes may be targeted by miR-1266 and miR-185, respectively, through binding to their 3′UTR regions.Transfection of PC3 and DU145 cells with miR-1266 and miR-185 induced apoptosis and reduced proliferation, whichalso revealed an inverse correlation with BCL2 and BCL2L1 gene expression in the treated cells. Conclusion: Ourdata suggests that miR-1266 and miR-185 may be novel candidates for further research in PCa treatment through theanti-apoptotic pathway.     

miR-124对非小细胞肺癌细胞增殖的影响及其机制

目的 探讨miR-124对非小细胞肺癌(NSCLC)细胞增殖的影响及其机制.方法 采用CCK8法和克隆形成实验评估miR-124对细胞增殖能力的影响.采用实时荧光定量PCR(qRT-PCR)和Western blot检测miR-124转染后细胞内PIM3的表达水平.通过双荧光素酶报告系统检测miR-124对PIM3荧光素酶活性的影响,证实miR-124可靶向作用于PIM3的3'UTR区.采用qRT-PCR和Western blot检测si-PIM3转染后细胞内PIM3的表达情况,采用CCK8法和克隆形成实验检测沉默PIM3后对细胞增殖能力的影响.结果 ①miR-124模拟物组的细胞活性(OD值)低于其对照组,差异有统计学意义,且呈时间依赖性;miR-124模拟物组的克隆形成数明显少于其对照组.②miR-124模拟物组的miR-124过表达后,其PIM3表达低于其对照组,差异有统计学意义.③含野生型结合位点(PIM3 3'UTR区完整片段)的miR-124模拟物组的荧光素酶活性低于其对照组,差异有统计学意义;含突变型结合位点(PIM3 3'UTR区突变片段)的两组无明显差异.④si-PIM3组的PIM3被沉默后,其表达水平明显低于其对照组,差异有统计学意义;si-PIM3组的细胞活性(OD值)低于其对照组,差异有统计学意义,且呈时间依赖性;si-PIM3组的克隆形成数明显少于其对照组.结论 miR-124可通过靶向作用于PIM3的3'UTR区下调后者的表达而抑制NSCLC细胞的增殖.     

MiR-150-5p Suppresses Colorectal Cancer Cell Migration and Invasion through Targeting MUC4

Growing evidence suggests that miR-150-5p has an important role in regulating genesis of various types of cancer. However, the roles and the underlying mechanisms of miR-150-5p in development of colorectal cancer (CRC) remain largely unknown. Transwell chambers were used to analyze effects on cell migration and invasion by miR-150-5p. Quantitative real-time PCR (qRT-PCR), Western blotting and dual-luciferase 3’ UTR reporter assay were carried out to identify the target genes of miR-150-5p. In our research, miR-150-5p suppressed CRC cell migration and invasion, and MUC4 was identified as a direct target gene. Its effects were partly blocked by re-expression of MUC4. In conclusiomn, miR-150-5p may suppress CRC metastasis through directly targeting MUC4, highlighting its potential as a novel agent for the treatment of CRC metastasis.     

易瑞沙治疗化疗失败的晚期非小细胞肺癌

目的观察易瑞沙治疗化疗失败晚期非小细胞肺癌的疗效和不良反应。方法易瑞沙每天口服250mg治疗化疗失败的30例晚期非小细胞肺癌,1个月以后进行疗效评价,无进展者继续服用,之后每个月行CT检查评价疗效并临床密切观察,病情进展或不能耐受相关毒性者则停止使用易瑞沙。结果30例均可评价疗效,无CR者,PR6例(20.0%),SD14例(46.7%),PD10例(33.3%)。有效率(CR PR)为20.0%,疾病控制率(CR PR SD)66.7%,全组中位无进展生存期为3.0个月(0.7～29.0个月),中位生存期6.4个月(1.7～39.0个月)。主要不良反应包括皮疹22例,腹泻2例。没有患者因毒性不能耐受而停药。结论易瑞沙对化疗失败的晚期非小细胞肺癌具有一定疗效,不良反应轻微。     

前列腺癌中外周血miR-223-3p和miR-223-5p的表达水平及与临床病理特征和预后的相关性

目的探究外周血miR-223-3p和miR-223-5p水平与前列腺癌患者的临床病理特征和预后的关系。方法选取前列腺癌患者64例,RT-PCR检测患者血清miR-223-3p和miR-223-5p水平,多因素logistic回归分析前列腺癌患者预后的危险因素。结果患者年龄、术前PSA差异无统计学意义(P>0.05);血清miR-223-3p、miR-223-5p表达水平与Gleason评分、有无骨转移、TNM临床分期有关(P<0.05)。存活组与死亡组PCa患者的基线特征比较,死亡组的血清miR-223-3p、miR-223-5p水平显著高于存活组(P<0.05),Gleason评分≥7、有骨转移、TNM临床分期Ⅲ+Ⅳ期的患者存活率显著低于Gleason评分<7、无骨转移、Ⅰ+Ⅱ期患者(P<0.05)。多因素logistic回归分析显示,miR-223-3p、miR-223-5p水平上升为影响脑膜瘤患者预后的独立危险因素(P<0.05)。结论血清miR-223-3p和miR-223-5p水平与前列腺癌患者临床病理特征和预后相关。     

MCM5在非小细胞肺癌中的表达及其临床意义

目的探讨MCM5在非小细胞肺癌（non-small cell lung cancer,NSCLC）中的表达及与其临床病理特征的关系。方法采用免疫组织化学SP法,检测78例原发性非小细胞肺癌组织、40例正常肺组织中MCM5的表达水平。结果非小细胞肺癌组MCM5阳性率高于正常肺组织组（P=0.000）。Ⅰ、Ⅱ、Ⅲ期非小细胞肺癌组织中MCM5阳性率比较,差异有统计学意义（P=0.031）;淋巴结转移者MCM5表达率与无淋巴结转移者比较也有统计学差异（P=0.015）。结论 MCM5可能参与了非小细胞肺癌的发生和发展过程;可能成为判断非小细胞肺癌发生发展和评价预后的有效参考指标之一,对于非小细胞肺癌的早期预测可能具有重要的临床意义。