Title: Changes in the expression of several apoptosis-regulated genes in apoptosis of human neuroblastoma cell line SH-SY5Y induced by 2,3-dioxoindoline
Abstract: Objective:To investigate the molecular mechanism underlying the effects of 2,3-dioxoindoline(Isatin,ISA) on the apoptosis of human neuroblastoma cell line.Methods: Human neuroblastoma cell line SH-SY5Y cells were divided into 4 groups: control group and 3 treatment groups treated with ISA(100,200,and 400 μmol/L) for 48 h.Fluorescent staining and DNA ladder analysis were used to examine the apoptosis of SH-SY5Y cells.Reverse transcription-polymerase chain reaction(RT-PCR) was used to mea-sure the mRNA expressions of VEGF,bcl-2,bax,sruvivin,and p53;flow cytometry was used to detect the protein expression of caspase 3.Results: When treated with 400 μmol/L ISA for 48 h,SH-SY5Y cells showed the typical apoptotic morphologic changes including chromatin clumping,nuclear fragmentation,and chromatin condensation;soft agar electrophoresis detected the obvious DNA ladder changes after 400 μmol/L ISA treatment(P0.05).RT-PCR showed that along with the increase in the concentration of ISA,the mRNA expression levels of bcl-2,VEGF,and survivin were all decreased(P0.05) but the expression of bax remained unchanced(P0.05),while the expression of p53 was increased in a concentration-dependent manner(P0.05).The positive rates of activated caspase-3 in SH-SY5Y cells were also significantly increased to 18.38%,26.93%,and 35.66% after treatment with ISA(100,200,and 400 μmol/L) compared with that in control group(11.23%,P0.05).Conclusion:ISA significantly induces apoptosis of human neuroblastoma SH-SY5Y cells,the action mechanism may be associated with the up-regulating p53 transcription and down-regulating bcl-2 and survivin transcription and increasing the intracellular level of active caspase-3.ISA is found to inhibit VEGF transcription,which is very important to suppress the growth and metastasis of tumor.
Publication Year: 2009
Publication Date: 2009-01-01
Language: en
Type: article
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