Growth inhibition and apoptosis of human B-cell lymphoma in vitro and in vivo by Bcl-2 short hairpin RNA

Liu,Yuchun; Shen,Yongmei; Qin,Chenhao; Shi,Yizhen; Rong,Guang; Yu,Xilong

doi:10.3892/or.2012.2081

Growth inhibition and apoptosis of human B-cell lymphoma in vitro and in vivo by Bcl-2 short hairpin RNA

Authors:
- Yuchun Liu
- Yongmei Shen
- Chenhao Qin
- Yizhen Shi
- Guang Rong
- Xilong Yu
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Affiliations: Radioimmunoassay Center, the Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215004, P.R. China, Clinical Laboratory, the Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215004, P.R. China
Published online on: October 16, 2012 https://doi.org/10.3892/or.2012.2081
Pages: 244-252

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Abstract

Bcl-2 is overexpressed in various types of human tumors, including Burkitt's lymphoma, and it is involved in tumorigenesis and chemoresistance, therefore, it is regarded as a potential target of gene therapy. In this study, RNA interference using short hairpin RNA (shRNA)-mediated RNA interference was introduced into Burkitt's lymphoma Raji cells to validate its effects on Bcl-2 expression and cell proliferation in vitro and in vivo. We constructed two types of Bcl-2 shRNA plasmid (pGenesil-1-Bcl-2-1 and pGenesil-1-Bcl-2-2) and negative control shRNA plasmid (pGenesil-1-NC) and stably transfected them into Raji cells. The expression levels of Bcl-2 mRNA and protein were assayed by RT-PCR, flow cytometry and western blotting. Cell proliferation was determined by cell count assay. The antitumor activities and apoptosis of the two types of Bcl-2 shRNA plasmid were evaluated in BALB/c nude mice bearing Burkitt's lymphoma inoculated with Raji cells. The results showed that the expression levels of Bcl-2 mRNA and protein decreased, compared with either the pGenecil-1-NC or the untransfected cell group (P<0.05). The cell proliferation assay showed that Bcl-2 shRNA significantly inhibited the growth of Raji cells (P<0.01). Furthermore, the tumor growth of the Bcl-2 shRNA cell group was dramatically lower and smaller than that of the negative control or untransfected cell group (P<0.01). Bcl-2 protein expression in the untransfected and the pGenesil-1-NC group were markedly higher than that of the pGenesil-1-Bcl-2-1 and the pGenesil-1-Bcl-2 group by immunohistochemistry (both P<0.01) and the results using transmission electron microscopy showed that Bcl-2 shRNA significantly induced Raji cell apoptosis. Additionally, the inhibition effect of pGenesil-1-Bcl-2-1 was better than that of pGenesil-1-Bcl-2-2. It has been suggested that vector-based Bcl-2 shRNA could effectively reduce the expression of Bcl-2 and induce apoptosis and growth inhibition of Burkitt's lymphoma Raji cells. Vector-based Bcl-2 shRNA could be a potential gene therapeutic strategy against human Burkitt's lymphoma.

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