长链非编码RNA H19促进肝癌细胞增殖和侵袭

目的:肝癌是严重危害人类健康的一类恶性肿瘤,但其发病机制目前仍不明确.虽然长链非编码RNA的异常表达与多种肿瘤的发生密切相关,但在肝癌中的报导尚不多见.因此,本文将探讨长链非编码RNA H19在肝癌组织和正常组织中表达差异,进一步检测其对肝癌细胞增殖和侵袭活性的调控及其分子机制,为开发长链非编码RNA的临床诊断试剂提供实验依据.方法:收集20例肝癌手术患者的癌变组织和20例正常组织,通过real time PCR检测H19的表达差异.在转染或干扰掉该H19后,采用MTT的方法检测HepG2细胞的增殖活性,通过Transwell小室的方法检测HepG2细胞的侵袭活性.结果:real time PCR检测发现H19 mRNA在肝癌组织中高表达,而在正常组织中低表达.过表达H19导致HepG2细胞过度增殖,敲除该lncRNA,细胞增殖和侵袭活性被抑制.结论:H19在肝癌组织中的表达明显高于正常肝组织,并且过表达H19导致肝癌细胞增殖活性增加,而敲除H19会导致肝癌细胞的增殖和侵袭活性明显减弱.该LncRNA在肝癌组织中的过量表达有助于肝癌的临床诊断,同时H19在肝癌的恶性肿瘤生物活性中发挥着重要的作用,因此,H19是一个潜在的治疗肝癌的药物作用靶点,为开发新的抗肿瘤药物提供了新的治疗靶点.     

外泌体介导长链非编码RNA H19促进肝癌细胞增殖与转移

外泌体是由细胞分泌的直径为30～150 nm的小囊泡,含有丰富的mRNA、microRNA和长链非编码RNA(lncRNA)。目前,大多数外泌体研究都集中在mRNA和microRNA,而对lncRNA的生物学功能并不十分清楚。研究表明,肿瘤细胞外泌体 lncRNA H19在肿瘤细胞的增殖、迁移和侵袭中发挥了重要作用。本研究将筛选到的lncRNA H19高表达的肝癌细胞HCCLM3,分别收集其高表达lncRNA H19的外泌体和其下调lncRNA H19表达后的外泌体。然后,将收集到的外泌体分别添加到lncRNA H19低表达的肝癌细胞Hep3B和HepG2孵育液中。孵育24 h后,检测其对肿瘤细胞的增殖、迁移和侵袭能力的影响。结果显示,肝癌细胞HCCLM3可分泌大量的外泌体,且能被其他肿瘤细胞大量摄取;与下调lncRNA H19表达的外泌体相比,lncRNA H19高表达的外泌体能显著增强Hep3B和HepG2细胞的增殖、迁移和侵袭能力。而这一作用可通过激活PI3K/AKT/mTOR通路实现。上述结果表明,lncRNA H19高表达的肝癌细胞以外泌体方式,增强邻近肝癌细胞的增殖、迁移和侵袭能力,促进肝癌的发生与发展。     

Decreased miR-204 in H. pylori-Associated Gastric Cancer Promotes Cancer Cell Proliferation and Invasion by Targeting SOX4

Background

The molecular mechanism between Helicobacter pylori (H. pylori) infection and gastric cancer remained largely unknown. In this study, we determined the role of miRNA in H. pylori induced gastric cancer.

Methods and Results

We found that miR-204 was decreased in H. pylori positive tissues by qRT-PCR. Knockdown of miR-204 enhanced the invasion and proliferation ability of gastric cancer cells in vitro. Luciferase assay revealed that SOX4 was target gene of miR-204, which was found up-regulated in H. pylori positive tissues. Down-regulation of miR-204 and over-expression of SOX4 promoted epithelial-mesenchymal transition process.

Conclusion

Taken together, our findings demonstrated that miR-204 may act as a tumor suppressor in H. pylori induced gastric cancer by targeting SOX4.     

Overexpression of Long Non-Coding RNA HOTAIR Promotes Tumor Growth and Metastasis in Human Osteosarcoma

Human osteosarcoma usually presented a high tendency to metastatic spread and caused poor outcomes, however, the underlying mechanism was still largely unknown. In the present study, using a series of in vitro experiments and an animal model, we investigated the roles of HOX antisense intergenic RNA (HOTAIR) during the proliferation and invasion of osteosarcoma. According with our results, HOTAIR was commonly overexpressed in osteosarcoma, which significantly correlated with advanced tumor stage, highly histological grade and poor prognosis. In vitro and in vivo experiments demonstrated that knockdown of HOTAIR could notably suppress cellular proliferation, inhibit invasion and decrease the secretion of MMP2 and MMP9 in osteosarcoma. Collectively, our results suggested that HOTAIR might be a potent therapeutic target for osteosarcoma.     

MiR-196a促进人非小细胞肺癌细胞NCI-H1299侵袭转移的研究

目的:非小细胞肺癌发生、发展的分子机制仍是目前研究的热点与难点,新近研究表明microRNA在肿瘤的发展过程中起着重要的作用.本研究旨在探讨miR-196a在人非小细胞肺癌组织及细胞系中的表达水平,以及抑制miR-196a对非小细胞肺癌细胞侵袭转移能力的影响.方法:通过real-time PCR技术检测人非小细胞肺癌及细胞系中miR-196a的表达水平,通过转染miR-196a inhibitors抑制miR-196a的表达水平,并通过定量PCR检测转染效率.利用transwell实验检测下调miR-196a对NCI-H1299细胞的迁移、侵袭能力的影响.结果:相对于正常肺组织及细胞,在非小细胞肺癌组织和细胞中miR-196a的表达水平出现了显著的上调,NCI-H1299细胞中转染miR-196a inhibitors能显著抑制miR-196a的表达水平且抑制miR-196a的表达能降低NCI-H1299细胞的迁移、侵袭能力.结论:定量PCR结果显示miR-196a在非小细胞肺癌组织及细胞中表达显著上调,而封闭其表达能影响非小细胞肺癌细胞的迁移和侵袭功能,提示miR-196a的表达上调可能在非小细胞肺癌的发生、发展中起着关键的作用,并有可能作为将来非小细胞肺癌诊断、预后的分子靶标.     

LncRNA AC009686.2促进乳腺癌细胞增殖和侵袭

长非编码RNAs(long non-coding RNAs,LncRNAs)作为一类基因表达的调控因子,在多种肿瘤的发生发展中发挥关键作用,然而LncRNAs在乳腺癌中的作用及相关机制尚未完全阐明.为了寻找在乳腺癌发生发展中起关键作用的LncRNAs,本研究通过分析TCGA数据库发现,LncRNA AC009686.2...     

长链非编码RNAIFNG-AS1靶向miR-19b-1-5p调控oxLDL诱导的血管内皮细胞增殖、凋亡的机制研究

为了探讨长链非编码RNA干扰素活化基因的反义核糖核酸(lncRNA IFNG-AS1)对氧化型低密度脂蛋白(oxLDL)诱导的人脐静脉血管内皮细胞EVC-304增殖、凋亡的影响和调控机制,该研究采用100 μg/mL的oxLDL分别处理转染si-IFNG-AS1、miR-19b-1-5p mimics或共转染si-IF...     

LncRNA H19 Promotes Cell Proliferation,Migration, and Angiogenesis of Glioma by Regulating Wnt5a/β-Catenin Pathway via Targeting miR-342

Cellular and Molecular Neurobiology - Glioma is the most common malignant brain tumor and long non-coding RNAs (lncRNAs) have been reported to play an important role in the growth and angiogenesis...     

Long noncoding RNA H19 accelerates tenogenic differentiation by modulating miR-140-5p/VEGFA signaling

Rotator cuff tear (RCT) is a common tendon injury, but the mechanisms of tendon healing remain incompletely understood. Elucidating the molecular mechanisms of tenogenic differentiation is essential to develop novel therapeutic strategies in clinical treatment of RCT. The long non-coding RNA H19 plays a regulatory role in tenogenic differentiation and tendon healing, but its detailed mechanism of action remains unknown. To elucidate the role of H19 in tenogenic differentiation and tendon healing, tendon-derived stem cells were harvested from the Achilles tendons of Sprague Dawley rats and a rat model of cuff tear was established for the exploration of the function of H19 in promoting tenogenic differentiation. The results showed that H19 overexpression promoted, while H19 silencing suppressed, tenogenic differentiation of tendon-derived stem cells (TDSCs). Furthermore, bioinformatic analyses and a luciferase reporter gene assay showed that H19 directly targeted and inhibited miR-140-5p to promote tenogenic differentiation. Further, inhibiting miR-140-5p directly increased VEGFA expression, revealing a novel regulatory axis between H19, miR-140-5p, and VEGFA in modulating tenogenic differentiation. In rats with RTC, implantation of H19-overexpressing TDSCs at the lesion promoted tendon healing and functional recovery. In general, the data suggest that H19 promotes tenogenic differentiation and tendon-bone healing by targeting miR-140-5p and increasing VEGFA levels. Modulation of the H19/miR-140-5p/VEGFA axis in TDSCs is a new potential strategy for clinical treatment of tendon injury.Key words: lncRNA, miRNA, tendon stem cell, rotator cuff tear repair     

Inhibition of Glioma Cell Lysosome Exocytosis Inhibits Glioma Invasion

Cancer cells invade by secreting enzymes that degrade the extracellular matrix and these are sequestered in lysosomal vesicles. In this study, the effects of the selective lysosome lysing drug GPN and the lysosome exocytosis inhibitor vacuolin-1 on lysosome exocytosis were studied to determine their effect on glioma cell migration and invasion. Both GPN and vacuolin-1 evidently inhibited migration and invasion in transwell experiments and scratch experiments. There are more lysosomes located on the cell membrane of glioma cells than of astrocytes. GPN decreased the lysosome number on the cell membrane. We found that rab27A was expressed in glioma cells, and colocalized with cathepsin D in lysosome. RNAi-Rab27A inhibited lysosome cathepsin D exocytosis and glioma cell invasion in an in vitro assay. Inhibition of cathepsin D inhibited glioma cell migration. The data suggest that the inhibition of lysosome exocytosis from glioma cells plays an important modulatory role in their migration and invasion.     

miR-373 Inhibits Glioma Cell U251 Migration and Invasion by Down-Regulating CD44 and TGFBR2

Glioblastoma multiforme (GBM) is the most malignant glioma, unveiling the underlying mechanisms of its aggressiveness could promote the discovery of potential targets for effective treatment. MicroRNAs (miRNAs) are important participants in both development and disease, its involvement in cancers has long been recognized. In this study, we investigated the role of miRNA-373 (miR-373) in GBM cell line U251, demonstrated that although miR-373 does not affect cell growth of U251, it inhibits migration and invasion of U251. Forced expression of miR-373 down-regulates the expressions CD44 and TGFBR2, while knockdown of CD44 and TGFBR2 presents the similar phenotype as miR-373 overexpression, suggesting that CD44 and TGFBR2 are functional targets of miR-373, down-regulation of CD44 and TGFBR2 by miR-373 are partly responsible for the migration, and invasion suppressive role of miR-373 in U251.     

Up-Regulation of microRNA-183 Promotes Cell Proliferation and Invasion in Glioma By Directly Targeting NEFL

Glioblastoma multiforme (GBM) is the most common and lethal type of primary malignant brain tumor. In recent years, increasing reports suggest that discovery of microRNAs (miRNAs) might provide a novel therapeutical target for human cancers, including GBM. The expression and roles of microRNA-183 (miR-183) has been explored in several types of human cancers, including in GBM, and plays important roles in tumor initiation and progression. However, its biological functions in GBM remain largely unknown. In this study, we demonstrated that miR-183 was significantly up-regulated in astrocytoma tissues and glioblastoma cell lines. Introduction of miR-183 mimics into U251 cells could promoted, while its antisense oligos inhibited cell proliferation and invasion. Moreover, we identified neurofilament light polypeptide (NEFL) as a novel target gene of miR-183. The expression levels of NEFL are inversely correlated with that of miR-183 in human astrocytoma clinical specimens. In addition, NEFL-siRNA could significantly attenuate the inhibitory effects of knockdown miR-183 on the proliferation and invasion of U251 cells via mTOR signaling pathway. Overall, This study revealed that miR-183 promotes glioma cell proliferation by targeting NEFL, and also demonstrated that miR-183 could be a potential target for GBM treatment.     

Long Non-coding RNA HOXA10-AS Promotes Invasion and Migration of Hepatocellular Carcinoma Cells

Some studies have showed that long non-coding RNA (lncRNA) HOXA10-AS acts as an oncogene and regulates the invasion and metastasis of tumor cells. However, its mechanism in the invasion and migration of hepatocellular carcinoma (HCC) cells is unclear. The purpose of this study was to analyze the expression of HOXA10-AS in HCC tissues and its clinical significance, detect the influence of HOXA10-AS on the invasion and migration of HCC cells, and explore the mechanism of HOXA10-AS in promoting the invasion and migration of HCC cells. The results of quantitative real-time PCR (qRT-PCR) showed that the expression of HOXA10-AS was significantly upregulated in HCC tissues compared with the adjacent non-HCC tissues. Age and gender did not show significant correlation with HOXA10-AS expression, while tumor size, lymphatic metastasis and distant metastasis showed significant correlation with HOXA10-AS expression. Meanwhile, the expression of HOXA10-AS in HCC cells was higher than that in normal liver cells. After interfering with HOXA10-AS in HCC cell lines HepG2 and QGY7701, Transwell invasion and scratch experiments showed that the invasion and migration ability of HOXA10-AS cells in the HOXA10-AS group was significantly lower than that in the control group. Western blotting results showed that the expression levels of vimentin and N-cadherin were significantly lower than those of the control group, while the E-cadherin expression was significantly increased. The TGFβ1/Smads signaling pathway was inhibited after HOXA10-AS interference. In summary, HOXA10-AS promotes the invasion and migration of HCC cells by the TGFβ1/Smads signaling pathway.