Antiangiogenic effect of celastrol on the growth of human glioma: an in vitro and in vivo study
Authors: Yu-lun Huang
You-xin Zhou
Journal: Chinese Medical Journal (2009)
Discipline: CLINICAL MEDICINE
Keywords: Celastrol; anticancer activity; glioma; angiogenesis; VEGF
Affiliations: Department of Neurosurgery, the First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China. 

Abstract:  Background: Celastrol is a major active component of Tripterygium wilfordii named "Thunder God Vine", which is widely used to treat rheumatoid arthritis in China. The present study aims to demonstrate that celastrol has potent anticancer activity against glioma in vitro and in vivo. Methods: Proliferation, migration, and tube formation of ECV-304 cells were determined by MTT and matrigel assays. The antiangiogenesis effect of celastrol was assessed by the chick chorioallantoic membrane assay and the in vivo matrigel plug assay. Tumor microvessels (MVD) were determined immunohistochemically with anti-CD34 antibody. Vascular endothelial growth factor (VEGF) expression was defined as positive if distinct staining of the cytoplasm was observed in at least 10% of tumor cells at the deepest invasive site, central portion and superficial part of the tumor. MVD was estimated by averaging the counts of three times at a x 200 field in the most vascularized area of the deepest invasive site. Results: Celastrol purified from T. wilfordii inhibited the proliferation of vascular endothelial cells (ECV-304) with an IC50 value of 1.33 microg/ml. Celastrol, at the concentration of 0.2 microg/ml, significantly inhibited cell migration and tube formation. Celastrol inhibited angiogenesis in a dose-dependent manner both in vitro and in vivo. Subcutaneous administration of celastrol 5 days a week for 4 consecutive weeks significantly reduced tumor volume in a dose-dependent manner in the SHG-44 xenograft model. Celastrol at each different dose level lowered the density of MVD significantly in tumor bearing nude mice compared to the control group. Immunohistochemistry experiments further revealed that celastrol also decreased the level of VEGFR-1 and VEGFR-2 expression, but not the level of VEGF expression. Conclusions: Celastrol elicits antiangiogenic effects in vitro and in vivo, and could be of potential use in the treatment of malignant cancers such as glioblastoma. 

Retracted: No

PubPeer comments:
 Fig 2.  miR-17-3P promotes the survival of human colorectal cancer cells. SW480 and LoVo cells were transfected by miR-17-3P inhibitor, mimic or negative miRNA (C-MiR) ![file] Fig 2 from [Liu et al (2017)] .  miR-107 promotes the survival of human CRC cells. SW480 and LoVo cells were transfected by the miR-107 inhibitor, mimic, or negative miRNA (C-MiR). ![file] Two papers report identical results, except for the miRNA. , 
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 Discussion:  By **online bioinformatic prediction** and luciferase reporter assay, we confirmed that there is a binding site of miR-17-3P in Par4 3u2032-UTR, and Par4 is a direct target of miR-17-3p in human CRC cells. This seems to allude to a paragraph of Results:  **Par4 is the target of miR-17-3P** Par4 is a tumor suppressor in various cancers, including human CRC (11). We predicated the potential targets of miR-17-3P using miRDB and TargetScan databases, and found that there is possible binding site of miR-17-3P on 3u2032-UTR of Par4 (Fig. 3A). To identify the prediction and elucidate the underlying mechanism by which miR-17-3P regulates CRC cells survival, we conducted the luciferase reporter assay in SW480 and LoVo cells. As seen in Fig. 3A, the wild or mutant 3u2032-UTR of Par4 were cloned into luciferase vector, respectively. Luciferase activities of wild Par4-3u2032UTR were strongly reduced by miR-17-3P mimic, but there was no difference in the luciferase activities of mutant Par4-3u2032UTR in CRC cells (Fig. 3B). ![file] Liu et al (2017) have the same paragraph:  **Par4 Is the Target of miR-107** Par4 is a tumor suppressor in various cancers, including human CRC15. We predicted the potential targets of miR-107 using Microrna.org and TargetScan databases and found that there is a possible binding site of miR-107 in the 3u2032-UTR of Par4 mRNA (Fig. 3A). To identify the prediction and elucidate the underlying mechanism by which miR-107 regulates CRC cell survival, we conducted the luciferase reporter assay in SW480 and LoVo cells. The wild or mutant 3u2032-UTR of Par4 mRNA was cloned into the luciferase vector (Fig. 3A). The luciferase activity of wild Par4-3u2032UTR was strongly reduced by the miR-107 mimic, but there was no difference in the luciferase activity of mutant Par4-3u2032UTR in CRC cells (Fig. 3B). ![file] , 
 Here I have compared the two References sections, as another reminder that the same paper has been published twice with very little alteration. ![file] Other products of a papermill specialising in this style of paper are collated in a spreadsheet: https: docs.google.com spreadsheets d 19wUtpXazLg2zKbTpDBuq0smG6-A7VHYifkieoyyuTcQ edit?usp=sharing ,