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Bcl-2 is overexpressed in various types of human tumors, including Burkitt'slymphoma, and it is involved in tumorigenesis and chemoresistance, therefore,it is regarded as a potential target of gene therapy. In this study, RNA interferenceusing short hairpin RNA (shRNA)-mediated RNA interference was introduced intoBurkitt's lymphoma Raji cells to validate its effects on Bcl-2 expression andcell proliferation in vitro and in vivo. We constructed two types of Bcl-2 shRNAplasmid (pGenesil-1-Bcl-2-1 and pGenesil-1-Bcl-2-2) and negative control shRNAplasmid (pGenesil-1-NC) and stably transfected them into Raji cells. The expressionlevels of Bcl-2 mRNA and protein were assayed by RT-PCR, flow cytometry and westernblotting. Cell proliferation was determined by cell count assay. The antitumoractivities and apoptosis of the two types of Bcl-2 shRNA plasmid were evaluatedin BALB/c nude mice bearing Burkitt's lymphoma inoculated with Raji cells. Theresults 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&lt;0.05).The cell proliferation assay showed that Bcl-2 shRNA significantly inhibited thegrowth of Raji cells (P&lt;0.01). Furthermore, the tumor growth of the Bcl-2 shRNAcell group was dramatically lower and smaller than that of the negative controlor untransfected cell group (P&lt;0.01). Bcl-2 protein expression in the untransfectedand the pGenesil-1-NC group were markedly higher than that of the pGenesil-1-Bcl-2-1and the pGenesil-1-Bcl-2 group by immunohistochemistry (both P&lt;0.01) and theresults using transmission electron microscopy showed that Bcl-2 shRNA significantlyinduced Raji cell apoptosis. Additionally, the inhibition effect of pGenesil-1-Bcl-2-1was better than that of pGenesil-1-Bcl-2-2. It has been suggested that vector-basedBcl-2 shRNA could effectively reduce the expression of Bcl-2 and induce apoptosisand growth inhibition of Burkitt's lymphoma Raji cells. Vector-based Bcl-2 shRNAcould be a potential gene therapeutic strategy against human Burkitt's lymphoma.

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