Growth inhibition and apoptosis of human B-cell lymphoma in vitro and             in vivo by Bcl-2 short hairpin RNA.
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.
Radioimmunoassay Center, the Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215004, PR China.
Yuchun
Y
Liu
Yongmei
Y
Shen
Chenhao
C
Qin
Yizhen
Y
Shi
Guang
G
Rong
Xilong
X
Yu
D000818:Animals; D017209:Apoptosis; D015153:Blotting, Western; D049109:Cell Proliferation; D005434:Flow Cytometry; D005822:Genetic Vectors; D006801:Humans; D007124:Immunoenzyme Techniques; D066298:In Vitro Techniques; D016393:Lymphoma, B-Cell / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D034741:RNA, Small Interfering