Effects of shRNA‐mediated silencing of PSMA7 on cell proliferation and vascular endothelial growth factor expression via the ubiquitin‐proteasome pathway in cervical cancer
Authors: Yan-Nan Chen
Chen-Chen Ren
Ling Liu
Xiao-An Zhang
Li Yang
Guo-Mei Cheng
Hui Liu
Journal: Journal of cellular physiology (2018)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: PSMA7; cervical cancer (CC); proliferation; ubiquitin-proteasome pathway (UPP); vascular endothelial growth factor (VEGF).
Affiliations: Department of Obstetrics and Gynecology, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou, China., Department of Imaging, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou, China., Department of Oncology, The First Affiliated Hospital of Kunming Medical University, Kunming, China.

Abstract:  This study aims to evaluate the effects of PSMA7 silencing on cervical cancer (CC) cell proliferation and vascular endothelial growth factor (VEGF) expression through the ubiquitin-proteasome pathway. CC tissues (n = 43) and normal tissues (n = 27) were first collected from patients. Human CC cell line (SiHa) and human normal cervical epithelial cells (H8) were obtained and classified into the normal, blank, negative control (NC), PSMA7-shRNA1, and PSMA7-shRNA2 groups, respectively. In situ hybridization was used to detect the expressions of wild-type and mutant p53 proteins. Immunofluorescence assay was carried out to test the activity of 20S proteasomes. Reverse transcription quantitative polymerase chain reaction and Western blot analysis were both performed to determine the expressions of PSMA7, ubiquitin, P27, P53, and VEGF in sample tissues and cells. 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay was used to analyze cell proliferation rates, and flow cytometry was used to analyze the cell cycle and the apoptotic rate. Compared with normal tissues, CC tissues showed increased expression levels of PSMA7, ubiquitin, p53, VEGF as well as increased activity of 20S proteasomes but exhibited a decrease in p27 expression. Compared with the blank and NC groups, the PSMA7-shRNA1 and PSMA7-shRNA2 groups all had decreased expression levels of PSMA7, ubiquitin, p53, and VEGF as well as decreased cell proliferation, 20S proteasomes activity, and cell number in the S phase, increased p27 expression, cell apoptosis and cell number in the G0/G1 phase. Our study demonstrated that PSMA7 silencing can suppress CC cell proliferation and VEGF expression in addition to promoting cell apoptosis through inhibiting the UPP signaling pathway. 

Retracted: No

PubPeer comments:
 Fig 7B,  protein bands of expression of PSMA7, ubiquitin, p53, p27 and VEGF detected by Western blotting . ![file] The loading control is a versatile band. With an additional lane at the left, it appears in Fig 8(a) from [Liu et al (2018)] and Fig. 9A of [Zhou et al (2018)] . ![] Another sighting, though now identified as beta-Actin: Fig 3A from [Gong et al (2018)] . ![] Fig. 8E from [Qin et al (2018)] : ![] Of course the other bands in Fig 7B seem to come from the same limited gamut as these other figures. Interested readers can find more examples collected in a spreadsheet: https: docs.google.com spreadsheets d 1r0MyIYpagBc58BRF9c3luWNlCX8VUvUuPyYYXzxWvgY edit#gid=262337249 , 
 Fig 10A. Wobbly, overhanging, hand-drawn flow-cytometry cell-cycle diagrams are frequent in this oeuvre. ![file] , 
 Fig 10C. ![file] The colored boxes are to call attention to similar panels in Fig 7C of [Pan et al (2018)] , ![] and 10A of [Li et al (2017)] . ![] ,