Gold(III) porphyrin 1a inhibited nasopharyngeal carcinoma metastasis in vivo and inhibited cell migration and invasion in vitro

A physiologically stable gold compound, gold(III) meso-tetraphenylporphyrin (gold-1a), has been shown to be effective in inducing apoptosis and prolonging the survival of hepatocellular carcinoma (HCC)-bearing rats as well as inhibiting the tumor growth of mice bearing nasopharyngeal carcinoma (NPC), neuroblastoma and colon carcinoma. In this study, we showed that gold-1a prolonged the survival of NPC metastasis-bearing mice and inhibited intrahepatic and lung metastasis. Histologically, gold-1a markedly reduced tumor microvessel formation. Consistently, in in vitro studies, gold-1a inhibited migration and invasion of C666-1 human NPC cells. The data strongly support the use of gold(III) compounds to treat cancer metastasis.     

姜黄素对人鼻咽癌CNE-2Z细胞侵袭和转移的体外实验研究

背景与目的:鼻咽癌放射治疗后的局部复发和远处转移是患者死亡的主要原因之一。姜黄素(curcumin)是从姜科姜黄属植物姜黄(Curcuma longa)根茎中提取的一种酚性色素,具有抗菌、抗肿瘤及抗氧化作用。本研究旨在探讨姜黄素对人鼻咽癌CNE-2Z细胞侵袭和转移的影响,并探讨其可能的机制。方法:以10、20、40和80μmol/L姜黄素分别处理CNE-2Z细胞24、48 h后,MTT法检测其对细胞的生长抑制作用。应用Transwell小室进行人工重组基底膜(matrigel)侵袭和运动实验,观察姜黄素对CNE-2Z细胞侵袭和转移的影响。RT-PCR和Western blot分别检测不同浓度姜黄素作用后细胞中表皮生长因子受体(epidermal growth factor receptor,EGFR)表达水平的变化。结果:应用姜黄素处理后,人鼻咽癌CNE-2Z细胞生长受到抑制,且作用呈时效-量效依赖关系;同时细胞侵袭和迁移能力明显降低;EGFR基因和蛋白表达亦明显减弱。结论:姜黄素能够减弱人鼻咽癌CNE-2Z细胞的体外侵袭和转移能力,其机制可能与降低EGFR的表达有关。     

A suppressive role of ionizing radiation-responsive miR-29c in the development of liver carcinoma via targeting WIP1

Hepatocellular carcinoma (HCC) is the third most common cause of cancer-related deaths worldwide, and it has been linked to radiation exposure. As a well-defined oncogene, wild-type p53-induced phosphatase 1 (WIP1) plays an inhibitory role in several tumor suppressor pathways, including p53. WIP1 is amplified and overexpressed in many malignancies, including HCC. However, the underlying mechanisms remain largely unknown. Here, we show that low-dose ionizing radiation (IR) induces miR-29c expression in female mouse liver, while inhibiting its expression in HepG2, a human hepatocellular carcinoma cell line which is used as a model system in this study. miR-29c expression is downregulated in human hepatocellular carcinoma cells, which is inversely correlated with WIP1 expression. miR-29c attenuates luciferase activity of a reporter harboring the 3′UTR binding motif of WIP1 mRNA. Ectopic expression of miR-29c significantly represses cell proliferation and induces apoptosis and G1 arrest in HepG2. In contrast, the knockdown of miR-29c greatly enhances HepG2 cell proliferation and suppresses apoptosis. The biological effects of miR-29c may be mediated by its target WIP1 which regulates p53 activity via dephosphorylation at Ser-15. Finally, fluorescence in situ hybridization (FISH) and immunohistochemical analyses indicate that miR-29c is downregulated in 50.6% of liver carcinoma tissues examined, whereas WIP1 is upregulated in 45.4% of these tissues. The expression of miR-29c inversely correlates with that of WIP1 in HCC. Our results suggest that the IR-responsive miR-29c may function as a tumor suppressor that plays a crucial role in the development of liver carcinoma via targeting WIP1, therefore possibly representing a target molecule for therapeutic intervention for this disease.     

MicroRNA-10b induced by Epstein-Barr virus-encoded latent membrane protein-1 promotes the metastasis of human nasopharyngeal carcinoma cells

MicroRNA-10b (miR-10b) has been reported to facilitate the metastasis of breast cancer. However, little is known about the role of miR-10b in the metastasis of nasopharyngeal carcinoma (NPC). Here, we show that high levels of miR-10b expression in Epstein-Barr virus (EBV)-positive latent membrane protein-1 (LMP1)-expressing NPC cells, and its expression is down-regulated by silencing LMP1 or Twist. Induction of miR-10b over-expression in LMP1-silent C666-1 cells promoted significant wound healing and transmembrane invasiveness in vitro. More importantly, miR-10b over-expression promoted the metastasis of NPC and accelerated the death of tumor-bearing nude mice. These findings strongly suggest that miR-10b positively regulates the metastasis of NPC.     

MicroRNA-195 suppresses tumor cell proliferation and metastasis by directly targeting BCOX1 in prostate carcinoma

Elucidation of the downstream targets regulated by the metastasis-suppressive miRNAs can shed light on the metastatic processes in prostate cancer (PCa). We conducted microarray analyses and found that miR-195 was significantly decreased in metastatic PCa. Low miR-195 expression is an independent prognostic factor for poor biochemical recurrence-free and overall survival. Forced expression of miR-195 in PCa cells drastically inhibits proliferation, migration and invasion in vitro and inhibits tumor growth and metastasis in vivo. BCOX1 is identified as a direct target of miR-195 in PCa, and is found to be drastically increased in metastatic PCa. BCOX1 knockdown phenotypically copies miR-195-induced phenotypes, whereas forced expression of BCOX1 reverses the effects of miR-195. Collectively, this is the first report unveils that loss of miR-195 expression and thus uncontrolled BCOX1 upregulation might drive PCa metastasis.

Electronic supplementary material

The online version of this article (doi:10.1186/s13046-015-0209-7) contains supplementary material, which is available to authorized users.     

IFIT3对鼻咽癌CNE⁃2R细胞侵袭转移及EMT的影响

目的 探讨干扰素诱导的具有四肽重复序列3（interferon induced protein with tetratricopeptide repeats 3,IFIT3）在鼻咽癌CNE⁃2R细胞中的表达及其对细胞侵袭、转移和上皮⁃间质转化（epithelial⁃mesenchymal transition,EMT）能力的影响。方法 采用RT⁃qPCR和 Western blot检测IFIT3在鼻咽癌细胞CNE⁃2R和正常鼻咽上皮细胞NP69中的表达水平。鼻咽癌CNE⁃2R细胞分别感染IFIT3 shRNA的3个序列LV1、LV2、LV3,并设立阴性对照组（NC组）;利用RT⁃qPCR和 Western blot检测上皮表型E⁃cadherin及间质表型N⁃cadherin和Vimentin的表达,采用划痕实验及Transwell 实验检测细胞的迁移和侵袭能力。结果 IFIT3在鼻咽癌CNE⁃2R细胞中的mRNA和蛋白表达水平均较NP69细胞显著上调（均P<0.001）。IFIT3在LV1组、LV2组和LV3组中的mRNA和蛋白表达水平均低于NC组（均P<0.001）,且以LV2组和LV3组的表达水平最低。与NC组相比,LV2组和LV3组的细胞迁移率显著下降,迁移和侵袭细胞数显著减少,E⁃cadherin蛋白表达水平显著升高,N⁃cadherin和Vimentin蛋白表达水平显著降低（均P<0.001）。结论 IFIT3在鼻咽癌CNE⁃2R细胞中表达上调,沉默IFIT3后CNE⁃2R细胞的侵袭、转移能力以及EMT显著被抑制。      

MicroRNA profiling identifies MiR-195 suppresses osteosarcoma cell metastasis by targeting CCND1

Metastasis is a leading cause of mortality for osteosarcoma patients. The molecular pathological mechanism remains to be elucidated. In the previously study, we established two osteosarcoma cell lines with different metastatic potentials. Differential expressed genes and proteins regarding metastatic ability have been identified. MicroRNAs are important regulators in tumorigenesis and tumor progression. In this study, microRNA microarray was used to assess the differential expressed miRNAs level between these two cell lines. One of the top ranked miRNAs-miR-195 was identified highly expressing in lowly metastatic cells. It was showed that over-expression of miR-195 substantially inhibits migration and invasion of osteosarcoma cells in vitro and pulmonary metastasis formation in vivo. Meanwhile, CCND1 was identified as the target gene of miR-195 and further studied. More importantly, Using real-time PCR, we evaluated the expression of miR-195 and CCND1 in osteosarcoma samples from 107 frozen biopsy tissues and 99 formalin- or paraformalin-fixed, paraffin-embedded (FFPE) tissues. Results indicated lowly expressed miR-195 or highly CCND1 correlated with positive overall survival and their expression inverse relate to each other. In summary, our study suggests miR-195 function as a tumor metastasis suppressor gene by down-regulating CCND1 and can be used as a potential target in the treatment of osteosarcoma.     

MiR-20a inhibits cutaneous squamous cell carcinoma metastasis and proliferation by directly targeting LIMK1

Background MicroRNA-20a (miR-20a) plays a key role in tumorigenesis and progression. But its function is reverse in different kinds of malignant tumor, and its role and mechanism in cutaneous squamous cell carcinoma (CSCC) remains unclear. Object To determine the miR-20a’s roles in CSCC and confirm whether LIMK1 is a direct target gene of miR-20a. Methods First miR-20a and LIMK1 expression levels were detected in six pairs of CSCC tissues and corresponding normal skin by qRT-PCR. Then MTT assays and colony formation assays were performed to evaluate the impact of miR-20a on cell proliferation. In addition, scratch migration assays and transwell invasion assays were performed to check miR-20a’s effect on cell metastasis. Since LIMK1 (LIM kinase-1) was predicted as a target gene of miR-20a, the changes of LIMK1 protein and mRNA were measured by western blot and qRT-RCR methods after miR-20a overexpression. Moreover the dual reporter gene assay was performed to confirm whether LIMK1 is a direct target gene of miR-20a. Finally LIMK1 mRNA and miR-20a in other 30 cases of CSCC pathological specimens were determined and a correlation analysis was evaluated. Results The miR-20a significantly low-expressed in CSCC tissues compared with that in matched normal tissues while LIMK1 has a relative higher expression. MiR-20a inhibited A431 and SCL-1 proliferation and metastasis. Both of LIMK1 protein and mRNA levels were downregulated after miR-20a overexpression. The dual reporter gene assays revealed that LIMK1 is a direct target gene of miR-20a. Furthermore, qRT-PCR results of LIMK1 mRNA and miR-20a in 30 cases of CSCC pathological specimens showed miR-20a is inversely correlated with LIMK1 expression. Conclusion Our study demonstrated that miR-20a is involved in the tumor inhibition of CSCC by directly targeting LIMK1 gene. This finding provides potential novel strategies for therapeutic interventions of CSCC.     

miR-375调控PDGFC基因表达抑制肝癌血管生成

  目的  miR-375已被证明能够抑制肝癌细胞增殖、迁移和侵袭,促进肝癌细胞凋亡,本研究旨在探索miR-375对肝癌新生血管生成的影响及其分子机制。  方法  利用血管生成相关的功能实验探索miR-375对肝癌新生血管生成的影响;使用生物信息学方法预测miR-375潜在的与血管生成相关的功能靶基因;在肝癌细胞系中过表达和下调表达miR-375,验证miR-375对靶基因的调控作用,并利用双荧光素酶报告实验明确其分子机制;靶基因功能挽救实验明确miR-375通过调控靶基因的表达发挥抑制肝癌新生血管生成的作用。  结果  miR-375能够抑制肝癌新生血管生成;血小板源性生长因子C（PDGFC）是miR-375的潜在功能靶基因;miR-375能够直接作用于PDGFC基因mRNA 3'-非编码端（3'-UTR）而抑制PDGFC基因表达;miR-375能够通过抑制PDGFC基因表达抑制肝癌新生血管生成。  结论  miR-375能够调控PDGFC基因表达抑制肝癌新生血管生成。      

By downregulating TIAM1 expression,microRNA-329 suppresses gastric cancer invasion and growth

Gastric cancer (GC) is one of the most common malignant tumors worldwide. Emerging evidence has shown that abnormal microRNAs (miRNAs) expression is involved in tumorigenesis. MiR-329 was previously reported to act as a tumor suppressor or oncogene in some types of cancer. However, its function in gastric cancer (GC) is unclear. Here, we found that miR-329 was down-regulated in GC compared with adjacent controls. Enforced expression of miR-329 inhibited proliferation, migration and invasion of gastric cancer cells in vitro. We identified T lymphoma invasion and metastasis 1 (TIAM1) gene as potential target of miR-329. MiR-329 levels inversely correlated with TIAM1 expression in GC. Importantly, TIAM1 rescued the miR-329-mediated inhibition of cell invasion and proliferation. Finally, reintroduction of miR-329 significantly inhibited tumor formation of GC in the xenograft mice. Our findings suggest that miR-329 is a tumor suppressor and potential therapeutic target of GC     

MicroRNA-138 suppresses invasion and promotes apoptosis in head and neck squamous cell carcinoma cell lines

Metastasis is a critical event in the progression of head and neck squamous cell carcinoma (HNSCC). To identify microRNAs associated with HNSCC metastasis, six paired HNSCC cell lines with different metastatic potential were examined. Using microarrays, a panel of differentially expressed microRNAs was identified, including reduction of miR-138 in highly metastatic cells. Ectopic transfection of miR-138 suppressed cell invasion and led to cell cycle arrest and apoptosis. Knockdown of miR-138 enhanced cell invasion and suppressed apoptosis. Thus, our results suggested miR-138 acts as a tumor suppresser and may serve as a therapeutic target for HNSCC patients at risk of metastasis.     

miR-1284过表达对人胃癌SGC-7901细胞基因表达谱及侵袭转移的影响

背景与目的：人胃癌组织微小RNA表达谱芯片筛选研究报道miR-1284与胃癌淋巴结转移有关,但其在胃癌中的具体作用鲜见报道。该研究探讨miR-1284过表达对人胃癌SGC-7901细胞基因表达谱及侵袭转移的影响。方法：以慢病毒介导miR-1284过表达转染胃癌SGC-7901细胞为实验组(LV-miR-1284组),转染空载体慢病毒载体的细胞为阴性对照组(LV-NC-GFP组),未转染慢病毒载体的细胞为空白组(Control组)。采用荧光定量PCR检测各组细胞miR-1284的表达,芯片检测各组细胞差异表达基因,生物信息学预测miR-1284的靶基因,Transwell侵袭实验检测各组细胞侵袭能力,裸鼠皮下移植瘤模型检测各组细胞转移能力。结果：PCR结果显示,与LV-NC-GFP组和Control组比较,LV-miR-1284组细胞miR-1284表达明显增高;芯片结果显示,LV-miR-1284组有20个基因表达显著上调,17个基因表达显著下调;生物信息学预测miR-1284的靶基因有138个基因;Transwell侵袭实验结果显示,LV-miR-1284组侵袭细胞数为70.00±2.37,其细胞侵袭能力较LV-NCGFP组(168.67±4.55)和Control组(170.33±3.08)明显减弱;体内裸鼠实验结果显示,LV-miR-1284组肝脏转移率为14.29%,其细胞转移能力较LV-NC-GFP组(85.71%)和Control组(85.71%)明显减弱。结论：miR-1284过表达能抑制人胃癌SGC-7901细胞侵袭转移,其机制可能与调控SUMO1、JUN基因表达有关。     

MiR-125a suppresses tumor growth,invasion and metastasis in cervical cancer by targeting STAT3

MiR-125a has been characterized as a tumor suppressor in several cancers. However, the role of miR-125a in cervical cancer is unknown. In this study, we found the expression of miR-125a was downregulated in cervical cancer patients, and negatively correlated with the tumor size, FIGO stage, and preoperative metastasis. Kaplan-Meier analysis showed that miR-125a expression predicted favorable outcome for cervical cancer patients. Dual luciferase assays identified the STAT3 gene as a novel direct target of miR-125a. Functional studies showed that miR-125a overexpression significantly suppressed the growth, invasion and epithelial-mesenchymal transition (EMT) of cervical cancer cells both in vitro and in vivo via decreasing STAT3 expression. Moreover, miR-125a conferred to G2/M cell cycle arrest, accompanied by inhibition of several G2/M checkpoint proteins. Mechanistically, inactivation of miR-125a during cervical carcinogenesis was caused by HPV suppression of p53 expression. Clinically, STAT3, the expression of which, predicted poorer outcome, was inversely correlated with miR-125a in cervical cancer. These data highlight the importance of miR-125a in the cell proliferation and progression of cervical cancer, and indicate that miR-125a may be a useful therapeutic target for cervical cancer.     

MiR-23a sensitizes nasopharyngeal carcinoma to irradiation by targeting IL-8/Stat3 pathway

Radioresistance poses a major challenge in nasopharyngeal carcinoma (NPC) treatment, but little is known about how miRNA regulates this phenomenon. In this study, we investigated the function and mechanism of miR-23a in NPC radioresistance, one of downregulated miRNAs in the radioresistant NPC cells identified by our previous microarray analysis. We observed that miR-23a was frequently downregulated in the radioresistant NPC tissues, and its decrement correlated with NPC radioresistance and poor patient survival, and was an independent predictor for reduced patient survival. In vitro radioresponse assays showed that restoration of miR-23a expression markedly increased NPC cell radiosensitivity. In a mouse model, therapeutic administration of miR-23a agomir dramatically sensitized NPC xenografts to irradiation. Mechanistically, we found that reduced miR-23a promoted NPC cell radioresistance by activating IL-8/Stat3 signaling. Moreover, the levels of IL-8 and phospho-Stat3 were increased in the radioresistance NPC tissues, and negatively associated with miR-23a level. Our data demonstrate that miR-23a is a critical determinant of NPC radioresponse and prognostic predictor for NPC patients, and its decrement enhances NPC radioresistance through activating IL-8/Stat3 signaling, highlighting the therapeutic potential of miR-23a/IL-8/Stat3 signaling axis in NPC radiosensitization.     

微小RNA-375对鼻咽癌细胞侵袭迁移及JAK2／STAT3 信号通路的影响

目的 探讨微小RNA-375（miR-375）调节鼻咽癌细胞侵袭、迁移的作用及对Janus激酶2／信号转导与转录激活子3（JAK2／STAT3）信号通路的靶向调控。方法 选取鼻咽癌CNE-1细胞并采用Lipofectamine 2000脂质体分别转染miR-375模拟物（miR-375组）和miR-375 阴性对照（NC组）,同时设不行转染的CNE-1细胞为对照组,采用实时荧光定量 PCR（QPCR）检测转染48 h各组miR-375的表达情况;划痕实验和Transwell 小室实验分别检测各组细胞的迁移能力和侵袭能力变化;Western blotting检测JAK2／STAT3信号通路中JAK2、STAT3蛋白的变化情况。结果 QPCR检测显示,miR-375在miR-375组的表达量为10.74±1.21,高于NC组的1.09±0.14和对照组的1.12±0.17,差异均有统计学意义（P＜0.05）;miR-375组的细胞迁移率为（18.22±1.78）％,低于对照组的（43.67±2.60）％和NC组的（49.70±1.31）％,差异均有统计学意义（P＜0.05）。miR-375组的穿膜细胞数为（45.76±4.34）个,亦低于对照组的（144.98±3.65）个和NC组的（126.23±6.32）个,差异均有统计学意义（P＜0.05）;miR 375组JAK2、STAT3蛋白的表达水平分别为0.31±0.6和0.27±0.05,均低于对照组的0.54±0.05和0.41±0.06以及NC组的0.50±0.06和0.43±0.07（P＜0.05）,NC组和对照组以上指标的差异均无统计学意义（P＞0.05）。结论 上调miR-375表达可抑制CNE-1细胞的侵袭和迁移能力,可能与抑制JAK2／STAT3 信号通路的激活有关。     

miR-107靶向FoxM1对鼻咽癌细胞生长和侵袭及迁移影响

目的microRNA与鼻咽癌(nasopharyngeal carcinoma,NPC)发生发展密切相关,但miR-107在鼻咽癌中的作用机制仍不清楚。本研究检测miR-107在鼻咽癌组织中的表达水平,并初步探讨其对鼻咽癌细胞增殖、迁移和侵袭影响的分子机制。方法收集2013-02-02-2013-12-24于佛山市第一人民医院经病理活检确诊的86例NPC组织和42例慢性鼻咽炎组织。采用原位杂交技术检测组织中miR-107的表达。分析miR-107表达与鼻咽癌患者临床特征及5年生存率的关系。双荧光素酶实验检测miR-107和人叉头框蛋白M1(forkhead box M1,FoxM1)在鼻咽癌细胞中的相互作用。在鼻咽癌细胞系CNE-2中转染miR-107 mimics和mimics NC,并设立mock对照组,采用实时荧光定量PCR法检测FoxM1 mRNA的表达量,蛋白质印迹实验检测FoxM1蛋白的表达量。然后通过CCK8、划痕和Transwell小室法检测鼻咽癌细胞的增殖、迁移和侵袭能力。结果miR-107在鼻咽癌组织中的阳性表达率(15.12%,13/86)低于慢性鼻咽炎组织(83.33%,35/42),差异有统计学意义,χ^2=96.37,P<0.001;与T分期(χ^2=24.816,P<0.001)和N分期(χ^2=29.368,P<0.001)有关联,且鼻咽癌miR-107阳性患者的5年生存率高于miR-107阴性组,χ^2=6.380,P=0.043。miR-107靶向调控FoxM1的3′UTR,降低了荧光素酶的表达活性,F=363.600,P<0.001。miR-107 mimics组FoxM1mRNA(F=267.600,P<0.001)和蛋白表达水平(F=101.300,P<0.001)低于mimics NC和mock组。与mimics NC和mock组相比,miR-107 mimics组细胞的增殖(F=1192.000,P<0.001)和迁移、侵袭(F=210.600,P<0.001)能力均降低。结论miR-107在鼻咽癌患者中低表达,通过靶向抑制FoxM1的表达而抑制鼻咽癌细胞的增殖、迁移和侵袭能力。     

MiR-195 suppresses non-small cell lung cancer by targeting CHEK1

MiR-195 suppresses tumor growth and is associated with better survival outcomes in several malignancies including non-small cell lung cancer (NSCLC). Our previous study showed high miR-195 plasma levels associated with favorable overall survival of non-smoking women with lung adenocarcinoma. To further elucidate role of miR-195 in NSCLC, we conducted in vitro experiment as well as clinical studies in a cohort of 299 NSCLC samples. We demonstrated that miR-195 expression was lower in tumor tissues and was associated with poor survival outcome. Overexpression of miR-195 suppressed tumor cell growth, migration and invasion. We discovered that CHEK1 was a direct target of miR-195, which decreased CHEK1 expression in lung cancer cells. High expression of CHEK1 in lung tumors was associated with poor overall survival. Our results suggest that miR-195 suppresses NSCLC and predicts lung cancer prognosis.     

MiR-373 targeting of the Rab22a oncogene suppresses tumor invasion and metastasis in ovarian cancer

Metastasis is major cause of mortality in patients with ovarian cancer. MiR-373 has been shown to play pivotal roles in tumorigenesis and metastasis; however, a role for miR-373 in ovarian cancer has not been investigated. In this study, we show that the miR-373 expression is down-regulated in human epithelial ovarian cancer (EOC) and inversely correlated with clinical stage and histological grade. Ectopic overexpression of miR-373 in human EOC cells suppressed cell invasion in vitro and metastasis in vivo, and the epithelial–mesenchymal transition process. Silencing the expression of miR-373 resulted in an increased migration and invasion of EOC cells. Using integrated bioinformatics analysis, gene expression arrays, and luciferase assay, we identified Rab22a as a direct and functional target of miR-373 in EOC cells. Expression levels of miR-373 were inversely correlated with Rab22a protein levels in human EOC tissues. Rab22a knockdown inhibited invasion and migration of EOC cells, increased E-cadherin expression, and suppressed the expression of N-cadherin. Moreover, overexpression of Rab22a abrogated miR-373-induced invasion and migration of EOC cells. Taken together, these results demonstrate that miR-373 suppresses EOC invasion and metastasis by directly targeting Rab22a gene, a new potential therapeutic target in EOC.