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Silencing UHRF1 Inhibits Cell Proliferation and Promotes Cell Apoptosis in Retinoblastoma Via the PI3K/Akt Signalling Pathway

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Pathology & Oncology Research

Abstract

This study aimed to investigate the effect of silencing ubiquitin-like with PHD and RING finger domains 1 (UHRF1) on the proliferation and apoptosis of retinoblastoma (RB) cells and to clarify the molecular mechanism of the UHRF1 gene in the development of RB. Human RB WERI-Rb-1 cells were selected and assigned into a blank group (WERI-Rb-1 cells with no transfection), NC-shRNA group (WERI-Rb-1 cells infected with NC-shRNA virus) and UHRF1-shRNA group (WERI-Rb-1 cells infected with pGC-UHRF1-shRNA-LV-GFP# (39–1) virus). The mRNA and protein expression of UHRF1 was detected by RT-qPCR and Western blot analysis. The effect of silencing UHRF1 on the proliferation and apoptosis of WERI-Rb-1 cells was assessed by MTT assay, EdU assay, flow cytometry, and Hoechst staining. Furthermore, the expression of cell cycle-related factor (cyclin D1), apoptosis-related factors (caspase-9, Bcl-2 and Bax), and PI3K/Akt signalling pathway-related factors (p-PI3K, PI3K, p-Akt and Akt) were measured via Western blot analysis. The RNA interference plasmid UHRF1-shRNA was successfully constructed. After WERI-Rb-1 cells were infected with UHRF1-shRNA, decreased mRNA and protein expression of UHRF1 was found. WERI-Rb-1 cells infected with UHRF1-shRNA showed inhibited proliferative ability and increased apoptosis. In the UHRF1-shRNA group, more cells arrested at the G0/G1 phase and less cells at the S and G2/M phases. WERI-Rb-1 cells infected with UHRF1-shRNA had increased expression of caspase-9 and Bax and decreased expression of Bcl-2 expression and decreased levels of p-PI3K and p-Akt. In conclusion, our study demonstrated that silencing UHRF1 could inhibit the proliferation of RB cells and promote apoptosis. The mechanism may be caused by the downregulation of the proportion of Bcl-2/Bax expression and the promotion of the expression of caspase-9 through the PI3K/Akt signalling pathway.

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Authors and Affiliations

  1. Key Laboratory of Zoonosis, Ministry of Education Institutes, College of Veterinary Medicine, Jilin University, No. 5333 Xi’an Road, Changchun, 130062, People’s Republic of China

    Yang Liu & Zengshan Liu

  2. Department of Genetic Medicine Weill Medical College, Cornell University, New York, NY, 10065, USA

    Yang Liu

  3. Department of Colorectal and Stomach Cancer Surgery, Jilin Cancer Hospital, Changchun, 130062, People’s Republic of China

    Guodong Liang

  4. Changchun University of Chinese Medicine Affiliated Hospital, Changchun, 130062, People’s Republic of China

    Tingting Zhou

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Liu, Y., Liang, G., Zhou, T. et al. Silencing UHRF1 Inhibits Cell Proliferation and Promotes Cell Apoptosis in Retinoblastoma Via the PI3K/Akt Signalling Pathway. Pathol. Oncol. Res. 26, 1079–1088 (2020). https://doi.org/10.1007/s12253-019-00656-7

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