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

• Published in: Oncology reports Vol. 29; no. 1; pp. 244 - 252
• Main Authors: LIU, YUCHUN, SHEN, YONGMEI, QIN, CHENHAO, SHI, YIZHEN, RONG, GUANG, YU, XILONG
• Format: Journal Article
• Language: English
• Published: Greece D.A. Spandidos 01.01.2013; Spandidos Publications UK Ltd
• Subjects: Animals; Apoptosis; Bcl-2; Blotting, Western; Burkitt's lymphoma; Cell Proliferation; Enzymes; Flow Cytometry; Gene expression; Genetic Vectors; Humans; Immunoenzyme Techniques; In Vitro Techniques; Lymphoma; Lymphoma, B-Cell - genetics; Lymphoma, B-Cell - metabolism; Lymphoma, B-Cell - pathology; Male; Mice; Mice, Inbred BALB C; Mice, Nude; pGenesil-1 vector; Plasmids; Protein expression; Proteins; Proto-Oncogene Proteins c-bcl-2 - antagonists & inhibitors; Proto-Oncogene Proteins c-bcl-2 - genetics; Proto-Oncogene Proteins c-bcl-2 - metabolism; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; RNA polymerase; RNA, Messenger - genetics; RNA, Small Interfering - genetics; short hairpin RNA; Studies; Tumor Cells, Cultured; Tumors; United States > US
• ISSN: 1021-335X; 1791-2431
• DOI: 10.3892/or.2012.2081

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.