Effects of shRNA‐mediated silencing of PSMA7 on cell proliferation and vascular endothelial growth factor expression via the ubiquitin‐proteasome pathway in cervical cancer
| Autor: | Chenchen Ren, Xiaoan Zhang, Li Yang, Yan-Nan Chen, Hui Liu, Ling Liu, Guo-Mei Cheng |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
Adult
Vascular Endothelial Growth Factor A 0301 basic medicine Proteasome Endopeptidase Complex Physiology Clinical Biochemistry Uterine Cervical Neoplasms Apoptosis Young Adult 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Cell Line Tumor Humans RNA Small Interfering Cell Proliferation Ubiquitin Cell growth Chemistry Cell Cycle Checkpoints Cell Biology Middle Aged Cell cycle Molecular biology Gene Expression Regulation Neoplastic Vascular endothelial growth factor 030104 developmental biology Real-time polymerase chain reaction Proteasome Cell culture 030220 oncology & carcinogenesis Female RNA Interference Tumor Suppressor Protein p53 Signal transduction Cyclin-Dependent Kinase Inhibitor p27 Signal Transduction |
| Zdroj: | Journal of Cellular Physiology. 234:5851-5862 |
| ISSN: | 1097-4652 0021-9541 |
| DOI: | 10.1002/jcp.26408 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text