MicroRNA-200b Regulates Cell Proliferation, Invasion, and Migration by Directly Targeting ZEB2 in Gastric Carcinoma

Kurashige, Junji; Kamohara, Hidenobu; Watanabe, Masayuki; Hiyoshi, Yukiharu; Iwatsuki, Masaaki; Tanaka, Yuta; Kinoshita, Koichi; Saito, Seiya; Baba, Yoshifumi; Baba, Hideo

doi:10.1245/s10434-012-2217-6

Cited by 158 publications

(117 citation statements)

References 38 publications

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“…To determine whether miR-335 directly targets ZEB2, a dual-luciferase reporter assay was performed; it was revealed that miR-335 significantly decreased the luciferase activity ZEB2, a member of the zinc finger family, functions as a transcriptional repressor of E-cadherin (37). ZEB2 has been identified as upregulated in various types of human cancer, including breast, gastric, head and neck, hepatocellular, ovarian and non-small cell lung carcinoma, and glioma (38)(39)(40)(41)(42)(43)(44). ZEB2 was previously identified as upregulated in PTC cell lines compared with normal thyroid cancer cells (45).…”

Section: Discussionmentioning

confidence: 99%

MicroRNA‑335 is downregulated in papillary thyroid cancer and suppresses cancer cell growth, migration and invasion by directly targeting ZEB2

Kan

Yang

2017

Oncol Lett

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Abstract. MicroRNAs (miRs) are a group of short, endogenous, non-protein-coding and single-stranded RNAs that regulate gene expression by binding to the 3'-untranslated region (3'UTR) of mRNAs, which results in their degradation or translational repression. The aim of the present study was to investigate the expression and function of miR-335 in human papillary thyroid cancer (PTC). Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was performed to quantify the relative miR-335 expression levels in PTC tissues and cell lines. The effect of miR-335 on the proliferation, migration and invasion of PTC cells was assessed by an MTT assay, and transwell migration and invasion assays, respectively. Dual-luciferase reporter assays were employed to explore whether miR-335 directly targeted the 3'UTR of the potential target gene zinc finger E-box binding homeobox 2 (ZEB2). RT-qPCR and western blotting were adopted to assess the effect of miR-335 on the mRNA and protein expression of ZEB2. RT-qPCR revealed that miR-335 was downregulated in PTC tissues and cell lines. The MTT assay and transwell migration and invasion assays demonstrated that the overexpression of miR-335 significantly inhibited the proliferation, migration and invasion of PTC cells. ZEB2 was identified as a direct target of miR-335 with computational analysis, which was confirmed with a dual-luciferase reporter assay, RT-qPCR and western blotting. The knockdown of ZEB2 significantly inhibited the proliferation, migration and invasion of PTC cells, indicating that ZEB2 may be a functional target of miR-335. Taken together, these findings suggested that miR-335 functioned as a tumor suppressor and suppressed the growth and metastatic behavior of PTC cells by targeting ZEB2. IntroductionThyroid cancer is the most common subtype of endocrine malignancy, with 300,000 new cases per year, and ~40,000 mortalities per year, worldwide (1). The incidence of thyroid cancer has increased in recent decades (2). It can be categorized into four major histologic groups, including papillary thyroid cancer (PTC), follicular thyroid cancer, poorly differentiated carcinoma and anaplastic thyroid cancer (3). PTC accounts for 80-90% of all thyroid cancer cases (4).A number of factors have been demonstrated to be associated with PTC progression, including genetic factors, environmental exposure, epigenetic alteration, nodular disease of the thyroid and radiation exposure (5,6). The prognosis of PTC patients is associated with their age, tumor size, lymph node invasion and distant metastasis (7). Currently, the standard therapeutic treatment for PTC is complete thyroidectomy followed by radioiodine and levothyroxine therapy (8). However, ~10% of patients with PTC develop recurrence and metastasis, which are associated with poor prognosis (9). Thus, it is necessary to characterize the molecular mechanisms of PTC initiation and development, and to develop novel, targeted therapies for PTC.The abnormal expression of microRNAs (miRNAs/miRs) has been implicated in ...

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Section: Discussionmentioning

confidence: 99%

MicroRNA‑335 is downregulated in papillary thyroid cancer and suppresses cancer cell growth, migration and invasion by directly targeting ZEB2

Kan

Yang

2017

Oncol Lett

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“…This gene belongs to the micro RNA family 200 of non-coding RNAs involved in regulation of cancer progression. miR200b is deregulated in different cancers 24 and its aberrant down-expression is also known to enhance tumor invasion [25][26][27] . Furthermore, a prognostic value of miR-200b in different cancers has been shown [28][29][30] .…”

Section: E Study Of Mir-200b Expression For Ht1080 In Droplets Microfmentioning

confidence: 99%

Chromatin immunoprecipitation in microfluidic droplets: towards fast and cheap analyses

et al. 2017

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Genetic organization is governed by the interaction of DNA with histone proteins, and differential modifications of these proteins is a fundamental mechanism of gene regulation. Histone modifications are primarily studied through chromatin immunoprecipitation (ChIP) assays, however conventional ChIP procedures are time consuming, laborious and require a large number of cells. Here we report for the first time the development of ChIP in droplets based on a microfluidic platform combining nanoliter droplets, magnetic beads (MB) and magnetic tweezers (MT). The droplet approach enabled compartmentalization and improved mixing, while reducing the consumption of samples and reagents in an integrated workflow. Anti-histone antibodies grafted to MB were used as a solid support to capture and transfer the target chromatin from droplets to droplets in order to perform chromatin immunoprecipitation, washing, elution and purification of DNA. We designed a new ChIP protocol to investigate four different types of modified histones with known roles in gene activation or repression. We evaluated the performances of this new ChIP in droplet assay in comparison with conventional methods. The proposed technology dramatically reduces analytical time from a few days to 7 hours, simplifies the ChIP protocol and decreases the number of cells required by 100 fold while maintaining a high degree of sensitivity and specificity. Therefore this droplet-based ChIP assay represents a new, highly advantageous and convenient approach to epigenetic analyses.

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“…Although a particular miRNA that specifically regulates cancer cell intravasation and extravasation has not yet been identified, it is still believed that these steps may also be regulated by miRNAs. miR-200 family is recognized as a master regulator of the epithelial phenotype by post transcriptionally suppression of the expression of the ZEB1 and ZEB2 EMT-inducing transcription factors in breast [28] and gastric cancer [29]. Acting in the opposite direction, ZEB1 and ZEB2, which promote not only tumor cell dissemination, but also the tumorinitiating capacity, has been shown to transcriptionally repress miR-200 family members, thereby establishing a double negative feedback loop that causes the reinforcement of cells in either the mesenchymal or epithelial state [25].…”

Section: Mirnas and Metastasismentioning

confidence: 99%

microRNA: New Players in Metastatic Process

Triulzi¹,

Iorio²,

Tagliabue³

et al. 2013

Oncogene and Cancer - From Bench to Clinic

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MicroRNA-200b Regulates Cell Proliferation, Invasion, and Migration by Directly Targeting ZEB2 in Gastric Carcinoma

Abstract: miR-200b regulates ZEB2 expression and thus controls metastasis in gastric cancer.

Cited by 158 publications

References 38 publications

MicroRNA‑335 is downregulated in papillary thyroid cancer and suppresses cancer cell growth, migration and invasion by directly targeting ZEB2

MicroRNA‑335 is downregulated in papillary thyroid cancer and suppresses cancer cell growth, migration and invasion by directly targeting ZEB2

Chromatin immunoprecipitation in microfluidic droplets: towards fast and cheap analyses

microRNA: New Players in Metastatic Process

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