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Acta Biochimica et Biophysica Sinica 2009 41(2):131-136; doi:10.1093/abbs/gmn015
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© The Author 2009. Published by ABBS Editorial Office in association with Oxford University Press on behalf of the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences.

Small hairpin RNA targeting at Bcl-2 increases cytarabine-induced apoptosis in Raji cells

Dongmei He* and Baoying Fang

Institute of Hematology, Medical College of Jinan University, Guangzhou 510632, China

* Correspondence address. Tel/Fax: +86-20-85220262; E-mail: thedm{at}jnu.edu.cn


  Abstract

Bcl-2, a prominent member of the Bcl-2 family proteins, is responsible for the dysregulation of apoptosis and resistance to chemotherapy. In this study, we investigated whether small hairpin RNA (shRNA) targeting at Bcl-2 mRNA could enhance cytarabine (Ara-C)-induced apoptosis in Raji cells. Bcl-2 shRNA was transfected into Raji cells and the expression levels of Bcl-2 mRNA and protein were assayed by RT–PCR and immunofluorescence. Cell proliferation was determined by MTT assay. Apoptosis was determined by morphological observation and flow cytometric analysis. Our results show that expression levels of Bcl-2 mRNA and protein from Raji cells transfected with Bcl-2 shRNA decreased, compared with either negative control shRNA group or untransfected cells group (P < 0.05). Viability of cells transfected with Bcl-2 shRNA was less than the cells transfected with control shRNA and untransfected Raji cells, respectively (P < 0.05). Bcl-2 shRNA combined with Ara-C significantly inhibited the growth of cells (P < 0.05). There was no difference in cell survival between control shRNA/Ara-C combination and cells treated with Ara-C alone. Using Giemsa staining, cells treated with Bcl-2 shRNA plus Ara-C at 48 h displayed changes of apoptosis. Apoptotic rates of Raji cells treated with Bcl-2 shRNA combined with Ara-C significantly increased (P < 0.05), compared with either control shRNA/Ara-C combination or Ara-C-treated cells alone. Our results suggest that the shRNA against Bcl-2 mRNA could increase Ara-C-induced apoptosis in Raji cells.

Keywords    Bcl-2; small hairpin RNA; Raji cell; cytarabine; apoptosis

Received: September 29, 2008; Accepted: November 25, 2008
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