Overexpression Of ERβ Participates In The Progression Of Liver Cancer Via Inhibiting The Notch Signaling Pathway
Authors: Yahua Wu
Xufeng Zhou
Benyi Yi
Yiping Zhang
Lili Wang
Journal: OncoTargets and therapy (2019)
Discipline: CLINICAL MEDICINE
Keywords: ERβ; HepG2 and Huh7; Notch signaling pathway; liver cancer; proliferation and apoptosis.
Affiliations: Department of Biochemistry and Molecular Biology, Basic Medical College of Jiujiang University, Jiujiang City, Jiangxi Province 332000, People's Republic of China.

Abstract:  Purpose: This study aimed to explore the role of Estrogen Receptor-β (ERβ)-mediated Notch signaling pathway in the regulation of proliferation and apoptosis in liver cancer cells. Methods: HepG2 cells (Pbi-EGFP-ER) were transfected with ERβ that mediated by liposome, and normal HepG2 cells (Blank) and empty plasmid-transfected HepG2 cells (Pbi-EGFP-C) were used as controls. Then, Huh7 cells were transfected with shERβ lentivirus to knock down ERβ expression. The Huh7 cells were divided into three groups including Blank, experimental group (shERβ) and negative group (shLuc). Then, qRT-PCR, Western blot, CCK-8 assay, cell scratch assay, Transwell assay, Annexin V-FITC and PI double staining were performed based on these groups. Finally, a mouse xenograft model was constructed to verify the regulation of ERβ on Notch signaling pathway in liver cancer. Results: In HepG2 cells, the ERβ expression in Pbi-EGFP-E group was higher than that in Blank and Bi-EGFP-C group. Overexpression of ERβ inhibited HepG2 cell proliferation, migration, invasion and Ki67 protein expression, as well as promoted apoptosis, Bcl-2 and Bax expression. Overexpression of ERβ decreased Notch1, Notch2 and Hes1 expression. In Huh7 cells, the effect of low ERβ expression was contrary to that of high ERβ expression. The shERβ + DAPT group reversed the effect of shERβ on the volume and weight of transplanted tumors. Conclusion: ERβ may inhibit the development of liver cancer and promote apoptosis via inhibiting the Notch pathway. 

Retracted: Yes

PubPeer comments:
 Figure 4A shows 4 flow cytometry panels that appear to contain repetitive elements indicated by boxes drawn in the same color ![file] , 
 #1 Some of these panels appear to have crystallised, rather than emerging from a plotter in the usual manner. ![file] , 
  Figure 1 The transfection of Pbi-EGFP-ERu03b2 and shERu03b2. A, GFP-positive [**HepG2**] cells . Marked to show overlap with [below] Fig 1A from [Zhang et al (2019)] .  Fig. 1. Transfection efficiency in **MG-63** cell lines. (A) The expression and transfection efficiency of ERu03b2 were observed under fluorescence microscope . ![file] Could the authors clarify which cell-line and plasmids were used? , 
  Figure 3 Effect of ERu03b2 on HepG2 and Huh7 cell migration. ... (Cu2013D) Transwell assay used to detect the invasion of HepG2 and Huh7 cells . ![file] I have only examined one panel in any detail. ![file] , 
 Fig 3C and 3D also have several repetitive features ![file] , 
 6A.  The mice xenografts after 30 days of Huh7 xenograft model construction. ![file] The regularity of the shadows is what concern me here. Either the tumours were hovering above a featureless white background all at the same height, or the entire image is a digital construction. , 
 #6 More on 6A: Note that the highlights for the third specimen shows the light is coming from the upper right, so a shadow should be on the left, not right side. The lighting on the 1st and 2nd specimen is coming from the top-left. Inconsistent lighting effects on what is supposed to be a single picture. ,