Small hairpin RNA targeting at Bcl-2 increases cytarabine-induced apoptosis in Raji cells.
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.
He
Dongmei
D
Institute of Hematology, Medical College of Jinan University, Guangzhou, China. thedm@jnu.edu.cn
Fang
Baoying
B
D017209:Apoptosis / Q000187:drug effects* / Q000235:genetics*; D001483:Base Sequence; D045744:Cell Line, Tumor; D003561:Cytarabine / Q000494:pharmacology*; D017931:DNA Primers / Q000235:genetics; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D019254:Genes, bcl-2; D006801:Humans; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012334:RNA, Neoplasm / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection
D017931:DNA Primers; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D034741:RNA, Small Interfering; D003561:Cytarabine