Knockdown of long non‑coding RNA PVT1 inhibits the proliferation of Raji cells through cell cycle regulation

Zheng,Chanli; Xiao,Yu; Li,Yangqiu; He,Dongmei

doi:10.3892/ol.2019.10450

Knockdown of long non‑coding RNA PVT1 inhibits the proliferation of Raji cells through cell cycle regulation

Authors:
- Chanli Zheng
- Yu Xiao
- Yangqiu Li
- Dongmei He
View Affiliations

Affiliations: Institute of Hematology, School of Medicine, Jinan University, Guangzhou, Guangdong 510632, P.R. China
Published online on: June 7, 2019 https://doi.org/10.3892/ol.2019.10450
Pages: 1225-1234
Copyright: © Zheng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Long non‑coding RNA plasmacytoma variant translocation 1 (PVT1) has been reported to be associated with oncogenesis. However, the functional role of PVT1 in Burkitt lymphoma has not yet been addressed. The purpose of the present study was to investigate the effect of PVT1 knockdown by small interfering RNA (siRNA) on the proliferation of Burkitt lymphoma Raji cells and to explore its possible mechanism of action. An effective siRNA targeting PVT1 was screened and the corresponding short hairpin RNA (shRNA) was reconstructed into a lentiviral vector. Cell proliferation and cell cycle distribution were assessed by Cell Counting kit‑8 assay and flow cytometry, respectively. Protein expression levels of c‑Myc, cyclin‑dependent kinase inhibitor1A (CDKN1A, P21) and cyclin E1 (CCNE1) were detected by western blotting. A polymerase chain reaction (PCR) array was used to analyse the expression of genes associated with the cell cycle. PVT1 knockdown markedly suppressed proliferation, and induced cell cycle arrest at the G0/G1 phase in Raji cells. Protein expression levels of c‑Myc and CCNE1 were reduced, whereas P21 protein expression was markedly increased following downregulation of PVT1 in Raji cells. The cell cycle PCR array revealed that 54 genes were upregulated and 26 genes were downregulated in Raji cells following PVT1 knockdown. Reverse transcription‑quantitative PCR demonstrated that cyclin G2 (CCNG2), CDKN1A, Retinoblastoma‑like 2 (RBL2, p130), HUS1 checkpoint homolog, cyclin dependent kinase inhibitor 3 (CDKN3) and cyclin dependent kinase inhibitor 1B (CDKN1B) expression were upregulated, whereas the expression levels of CCNE1, cyclin D1 (CCND1) and cell division cycle 20 (CDC20) were downregulated in Raji cells with PVT1 knockdown. In conclusion, PVT1 knockdown may inhibit the proliferation of Raji cells by arresting cells in G0/G1 phase. Furthermore, inhibition of cell proliferation may be associated with a reduction inc‑Myc expression and alterations in the expression levels of cell cycle‑associated genes.

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