Abstract:

To observe the effect of 5-ALA photodynamic therapy (5-ALA　PDT) in combination with human telomerase reverse transcriptase mRNA antisense oligodeoxynucleotides (hTERT ASODN) on proliferation inhibition and apoptosis of human ovarian carcinoma 3AO cells,and to disscuse the mechanism of the killing effect of the  combination treatment on  human ovarian carcinoma 3AO cells.Methods MTT assay was used to screen he best appropriate combined parameters of different concentrations of 5-ALA and laser doses on proli
feration inhibition of human ovarian cancer 3AO  cells;the expression changes of  hTERT mRNA  were detected by RT-PCR after transfected with hTERT ASODN and sense oligodeoxynucleotides (SODN).Then human ovarian carcinoma 3AO cells were divided into blank control group,hTERT ASODN transfection group,hTERT SODN　transfection group,5-ALA PDT group,hTERT ASODN transfection combined with 5-ALA
PDT group,and  hTERT SODN transfection combined with 5-ALA PDT group.The inhibitory rates of  proliferation of  human ovarian carcinoma 3AO cells  we
re detected by MTT assay;the  apoptotic rates of  human ovarian carcinoma 3AO cells  were conducted with Annexin-V/PI and flow cytometry.Results The inhibitory rate of proliferation  of 5-ALA PDT on human ovarian carcinoma 3AO cells was dependent on the concentrations of photosensitizer and laser doses(P<0.05).The 24 h transfection in human ovarian carcinoma 3AO cells with different concentrations of hTERT ASODN diminished the abundance of hTERT mRNA in a concentration-dependent manner(P<0.05) and hTERT SODN had no effect on the expression of hTERT mRNA(P>0.05).The inhibitory rate of proliferation  of cells in hTERT ASODN transfection combined with 5-ALA PDT (0.50 mmol•L-1 5-ALA,2.50 J•cm-2 laser dose of PDT) group was  significantly increased compared with 5-ALA PDT group and hTERT ASODN transfection group(P<0.05).The apoptotic  rate of human ovarian carcinoma 3AO cells in hTERT ASODN transfection combined  with 5-ALA PDT group was 29.28%,which was significantly higher than those in  hTERT ASODN transfection group(12.79%) and 5-ALA PDT group (21.19%)(P<0.05).
Conclusion The  administration of 5-ALA PDT  combinated with hTERT ASODN display synergistic proliferation inhibition on human ovarian carcinoma 3AO cells,and the mechanism may be related to promoting apoptosis.
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Key words: 5-aminolevulinic acid, photodynamic therapy, human telomerase reverse transcriptase, combination treatment〖WT〗