MicroRNA-122-3p inhibits tumor cell proliferation and induces apoptosis by targeting Forkhead box O in A549 cells

Wen, Wang; Yang, Jinsong; Yu, Fenglei; Li, Wenjie; Wang, Li; Zou, Haoyu; Long, Xia

doi:10.3892/ol.2017.7577

Cited by 10 publications

(13 citation statements)

References 32 publications

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“…Liver specific miR-122 is exclusively non-vesicular, with key roles in hepatocyte growth, metabolism, and homeostasis 19 . miR-122 is also associated with human liver cancer metastasis and lung cancer 20 .…”

Section: Discussionmentioning

confidence: 99%

Extracellular small non-coding RNA contaminants in fetal bovine serum and serum-free media

Mannerström

Paananen

Abu-Shahba

et al. 2019

Sci Rep

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In the research field of extracellular vesicles (EVs), the use of fetal bovine serum (FBS) depleted of EVs for in vitro studies is advocated to eliminate the confounding effects of media derived EVs. EVdepleted FBS may either be prepared by ultracentrifugation or purchased commercially. Nevertheless, these preparations do not guarantee an RNA-free FBS for in vitro use. In this study we address the RNA contamination issue, of small non-coding (nc)RNA in vesicular or non-vesicular fractions of FBS, ultracentrifugation EV-depleted FBS, commercial EV-depleted FBS, and in our recently developed filtration based EV-depleted FBS. Commercially available serum-and xeno-free defined media were also screened for small ncRNA contamination. Our small ncRNA sequencing data showed that all EVdepleted media and commercially available defined media contained small ncRNA contaminants. Out of the different FBS preparations studied, our ultrafiltration-based method for EV depletion performed the best in depleting miRNAs. Certain miRNAs such miR-122 and miR-203a proved difficult to remove completely and were found in all media. Compared to miRNAs, other small ncRNA (snRNA, Y RNA, snoRNA, and piRNA) were difficult to eliminate from all the studied media. Additionally, our tested defined media contained miRNAs and other small ncRNAs, albeit at a much lower level than in serum preparations. our study showed that no media is free of small ncRNA contaminants. therefore, in order to screen for baseline RNA contamination in culturing media, RNA sequencing data should be carefully controlled by adding a media sample as a control. this should be a mandatory step before performing cell culture experiments in order to eliminate the confounding effects of media. Fetal bovine serum (FBS) contains essential factors required for cell growth, metabolism, attachment, and stimulation of proliferation 1. Thus, it is the most widely used supplement for culturing human and animal cells. One of the major concerns that has become evident recently is the presence of large amounts of extracellular vesicles (EVs) in FBS 2-4. Secreted by most cell types, EVs are mediators of cell-to-cell communication and immune regulation 5,6. They carry the genetic material (DNA and RNA) as well as proteins, lipids and other molecules, the transfer of which can alter the functions of the recipient cells. EVs present in FBS are co-isolated with cell-derived EVs and therefore act as contaminants that, affect the reliability of the read-out from cell culture experiments, as FBS-derived EVs are structurally and functionally similar to cell-derived EVs 7. In addition, FBS-EVs taken up by cultured cells cause substantial physiological effects 2. To overcome this problem, different methods are used to deplete EVs from FBS. Ultracentrifugation (UC) is commonly used, but this method removes EVs only partially 3,4,8. Commercially available depleted FBS are produced using proprietary protocols to reach higher purity levels. Also, commercially available defined, serum-free and animal pr...

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Section: Discussionmentioning

confidence: 99%

Extracellular small non-coding RNA contaminants in fetal bovine serum and serum-free media

Mannerström

Paananen

Abu-Shahba

et al. 2019

Sci Rep

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“…By surrounding targeted tissues with pro-tumorigenic signals, cancer cell-derived sEVs can help in priming the pre-metastatic niche [67][68][69][70]. Only one miRNA with tumor suppressive functions (miR-122-3p) was found to be associated with sEVs [71]. As discussed above, tumor cells may release such miRNAs in the extracellular space to increase their ability to survive.…”

Section: Discussionmentioning

confidence: 98%

Circulating miRNAs in Small Extracellular Vesicles Secreted by a Human Melanoma Xenograft in Mouse Brains

Guglielmi

Nardella

Musa

et al. 2020

Cancers

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The identification of liquid biomarkers remains a major challenge to improve the diagnosis of melanoma patients with brain metastases. Circulating miRNAs packaged into tumor-secreted small extracellular vesicles (sEVs) contribute to tumor progression. To investigate the release of tumor-secreted miRNAs by brain metastasis, we developed a xenograft model where human metastatic melanoma cells were injected intracranially in nude mice. The comprehensive profiles of both free miRNAs and those packaged in sEVs secreted by the melanoma cells in the plasma demonstrated that most (80%) of the sEV-associated miRNAs were also present in serum EVs from a cohort of metastatic melanomas, included in a publicly available dataset. Remarkably, among them, we found three miRNAs (miR-224-5p, miR-130a-3p and miR-21-5p) in sEVs showing a trend of upregulation during melanoma progression. Our model is proven to be valuable for identifying miRNAs in EVs that are unequivocally secreted by melanoma cells in the brain and could be associated to disease progression.

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“…As important genes in the post-transcriptional regulation system, miRNAs have attracted much attention in the medical field around the world 24–26. miRNAs are involved in a wide range of biological processes 27.…”

Section: Discussionmentioning

confidence: 99%

<p>kshv-mir-k12-1-5p promotes cell growth and metastasis by targeting SOCS6 in Kaposi’s sarcoma cells</p>

Zhang

Xiong

2019

CMAR

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Background: Kaposi’s sarcoma (KS) is a highly disseminated angiogenic tumour of endothelial cells. Many deregulated miRNAs, including kshv-mir-k12-1-5p, have been identified in KS. kshv-mir-k12-1-5p plays important roles in KS. However, the underlying mechanism is not fully understood. The aim of this study was to investigate the exact functions of kshv-mir-k12-1-5p in KS cells. Materials and methods: The biological functions of kshv-mir-k12-1-5p were studied using CCK-8, apoptosis, migration and invasion assays. Bioinformatics software was used to identify the target gene (SOCS6) of kshv-mir-k12-1-5p. A dual luciferase assay, Western blot (WB) and quantitative real-time polymerase chain reaction (q-PCR) were performed to further verify the target gene. The underlying molecular mechanisms of kshv-mir-k12-1-5p in KS cells were also explored. Results: kshv-mir-k12-1-5p can promote the proliferation, migration and invasion of KS cells and inhibit cell apoptosis. Suppressor of cytokine signalling 6 (SOCS6) was identified as a direct target of kshv-mir-k12-1-5p, and kshv-mir-k12-1-5p can downregulate SOCS6 expression. In addition, knockdown of SOCS6 rescued the effects of kshv-mir-k12-1-5p inhibitor. Hence, a direct relationship between kshv-mir-k12-1-5p and SOCS6 was confirmed. Conclusions: kshv-mir-k12-1-5p promotes the malignant phenotype of KS cells by targeting SOCS6, suggesting that kshv-mir-k12-1-5p could be a potential therapeutic target for KS.

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MicroRNA-122-3p inhibits tumor cell proliferation and induces apoptosis by targeting Forkhead box O in A549 cells

Cited by 10 publications

References 32 publications

Extracellular small non-coding RNA contaminants in fetal bovine serum and serum-free media

Extracellular small non-coding RNA contaminants in fetal bovine serum and serum-free media

Circulating miRNAs in Small Extracellular Vesicles Secreted by a Human Melanoma Xenograft in Mouse Brains

<p>kshv-mir-k12-1-5p promotes cell growth and metastasis by targeting SOCS6 in Kaposi’s sarcoma cells</p>

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