ZIC2 promotes viability and invasion of human osteosarcoma cells by suppressing SHIP2 expression and activating PI3K/AKT pathways
Authors: Anmin Jin
Shuaihao Huang
Journal: Journal of cellular biochemistry (2018)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: PI3K/AKT; SHIP2; ZIC2; metastasis; osteosarcoma
Affiliations: Department of Orthopedics, Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong, China.

Abstract:  Osteosarcoma is a malignant tumor of the skeletal system. The zinc finger transcription factor ZIC2 has been reported to be highly expressed in human cancers. The present study evaluated the effects of ZIC2 and the possible underlying mechanisms in the human osteosarcoma cells. The expression levels of ZIC2 in human fetal osteoblastic cell line (hFOB1.19), osteosarcoma cell lines (U-2OS, SaoS2, and MG63), normal bone tissue, and osteosarcoma tumor were analyzed by Western blot, and real-time quantitative RT-PCR (qRT-PCR). Osteosarcoma cells with either overexpressed ZIC2 or suppressed ZIC2 were analyzed to determine cell viability, colony formation, and cell invasion. The expressions of SHIP2 and PI3K/AKT signal pathway-related proteins were analyzed by Western blot and qRT-PCR. We first showed that ZIC2 is highly expressed in osteosarcoma cells and tissues. Then we demonstrated that overexpression of ZIC2 promoted viability, migration, and invasion of osteosarcoma cells, whereas suppression of ZIC2 showed opposite effects. Furthermore, SHIP2 expression was negatively regulated by ZIC2. Importantly, ZIC2 overexpression activated the PI3K/AKT signal pathway; however, overexpressed SHIP2 inhibited these effects. Lastly, we showed that activation of the PI3K/AKT signal pathway is essential for the effects of ZIC2 on osteosarcoma cells, as the effects of ZIC2 on the osteosarcoma cells were reversed by a PI3K/AKT inhibitor. Overall, ZIC2 is highly expressed in osteosarcoma cells and tissues, and its overexpression promotes viability, invasion of osteosarcoma cells via SHIP2 suppression, and PI3K/AKT activation. Thus, ZIC2 can be considered as a novel drug target for osteosarcoma management. 

Retracted: Yes

PubPeer comments:
 The Western blots in this paper have an interesting appearance that appears to be shared with blots in other papers from other research groups, such as these papers published in the Journal of Cellular Biochemistry: * Ying Liu et al. 2019, DOI: 10.1002 jcb.29568 * Long Liu et al. 2020, DOI: 10.1002 jcb.29642 * Lin Zhang et al. 2019, DOI: 10.1002 jcb.28729 * Yugeng Li et al. 2019, DOI: 10.1002 jcb.29146 * Xiaoliang Yang et al. 2019, DOI: 10.1002 jcb.28829 * Ning Han et al. 2019, DOI: 10.1002 jcb.29638 All these papers share particular characteristics, such as similar title structure, similar graph design, and the appearance of the Western blots. Western blot panels within the same figure, and across figures and papers, appear to share the same background or parts of the background. In addition, the bands are regularly spaced, all have similar rounded edges, without the usual smudges and specks. The bands also sometimes have a particular fish-like shape that resembles jumping sardines. For example, here is Figure 5C. Although the bands are differently shaped, the background of these panels is more similar than expected. Colored panels draw attention to some of those background areas that look unexpectedly similar. Other Western blot photos in this paper share the same phenomenon. Could the authors provide the original, uncropped blots, please? ![file] , 
 If the corresponding author opts for  BrendaWillingham12192 as an email address, this camphor-leafed waxy plant is in no position to criticize. There is the problem that Gmail is not accessible in mainland China, so I do not know if Dr Bik s request for original blots will reach its target. ![file] , 
 #1 The same background to bands is recognisable from Fig 7 of [Ma et al (2018)] - here at left. ![file] , 
 Below, in the lower half, are Figs 2C D and 3C D, nominally Colony formation and Cell invasion images for osteosarcoma cell-line cells MG63 under various conditions of tranfection (or not) with oeZIC2 or siZIC2. Above are Figs 2C and 2D from [Tian et al (2018)] :  Subsequently, pEGFP-H3, pEGFP-K-RasG12V T35S plus pEGFP-H3, or pEGFP-K-RasG12V T35S plus pEGFP-H3K9Q (2u2009g) was transfected into cell, and **c number of colonies** and **d cell migration** were respectively determined by colony formation assay and transwell assay. Also, Fig 4D:  migration . ![file] Evidently these were not in the initially-submitted version of the latter paper, as we read in the [Open Peer Review records] that a reviewer requests  Representative pictures for transwell migration and soft-agar colony formation assays . The reviewer was presumably satisfied, and did not notice the overlap between panels of 2D and 4D (green boxes), casting doubt on their identification. Overlaps between the two papers are marked. Also colony image reuse (after rotation). There are no authors in common. , 
 Retracted [23 September 2020] . ------------------------------------------------------------------  ZIC2 promotes viability and invasion of human osteosarcoma cells by suppressing SHIP2 expression and activating PI3K AKT pathways,u201d by Shuaihao Huang and Anmin Jin, J Cell Biochem. 2018; 2248u20102257: The above article, published online on 31 August 2017 in Wiley Online Library , has been retracted by agreement between the authors, the journal s Editoru2010inu2010Chief, Prof. Dr. Christian Behl and Wiley Periodicals LLC. The retraction has been agreed following an investigation based on allegations raised by a third party. Raw experimental data was not accessible anymore and thus the validity of the overall results could not be confirmed. ,