RT Journal Article
SR Electronic
T1 The Effect of Overexpression of hTERT on Etoposide (VP-16)-lnduced Apoptosis in Raji Cells
JF Chinese Journal of Clinical Oncology
JO Cancer Biol Med
FD China Anti-Cancer Association
SP 622
OP 626
VO 2
IS 3
A1 Dongmei He
A1 Huan Zhang
A1 Gexiu Liu
A1 Wei Yu
YR 2005
UL http://www.cancerbiomed.org/content/2/3/622.abstract
AB OBJECTIVE To explore whether overexpression of human telomerase reverse transcriptase (hTERT) in Raji cells can protect against etoposide (VP-16)-induced apoptosis.METHODS A lipofectin-mediated gene transfection method was used to transfer the hTERT gene into Raji cells. The polymerase chain reaction enzyme-linked immunoassay was employed to determine telomerase activity. The expression levels of hTERT protein were assayed by immunofluorescence using a fluoresce isothiocyanate label. Cell viability was determined using the trypan blue dye exclusion assay. Apoptosis was assessed by morphological observation and flow cytometry analysis.RESULTS The results showed that there was a marked increase in both mean fluorescence intensity of hTERT-protein-positive cells and telomerase activity in hTERT-transfected Raji cells (P<0.05), but there was no difference in hTERT protein and telomerase activity levels between Raji cells and vector-transfected Raji cells (P >0.05). There were more viable cells at 48 h and 72 h after treatment of hTERT-transfected Raji cells with 10 μmol/L VP-16 compared to either vector-transfected Raji cells and Raji cells (P <0.05). Apoptosis rates at 72 h after treatment with 10 μmol/L VP-16 were 4.34 ±1.03% in hTERT-transfected Raji cells, 33.21 ± 3.12% in vector-transfected Raji cells, and 31.63 ± 3.06% in Raji cells. There was a significant difference in the percentage of apoptotic cells between hTERT-transfected Raji cells and either vector-transfected Raji cells or Raji cells (P<0.05).CONCLUSION Overexpression of telomerase by transfection of hTERT gene can protect against etoposide-induced apoptosis in Raji cells.