Th e Effect of RNA Interference Silencing ERCC1 on Proliferation And Apoptosis of Lun g Canc er A549 Cell
S ong Ying
Qiu Qin-wei
HeZ hi-min
Gu Yi-xue
Abstract：A bstract: Objce tive bjective: To investigate the effection of excision repair cross complementation 1 ( ERCC1) downr-egulated by RNA interference on proliferation and apoptosis in lung cancer A 549 cell line.Furthermore, to study the chemotherapy sensitivity of A549 cells with down-regulation ERCC1 to cisplatin.Methods:The negative control interfering RNA(NC) together with three small interfering RNAs (S1, S2 and S3) targeting ERCC1 gene was designed and synthe-sized.The siRNAs (S 1, S2, S3, S1+S2+S3 and NC) were transfected to A549 cell with lipofectamine 2000, respective-ly.The mRNA and protein expression levels of ERCC 1 were evaluated by real-time RT-PCR and western blot assay .All three siRNAs(S1, S2 and S3) could decrease the expression of ERCC1, but S1+S2+S3 reduced ERCC1 mostly.SiRNAs NC and S1+S2+S3 were transfected into A549 cell in 96 well-plot, the concentration of 0, 0.5, 1, 2, 4, 8μg/mL cDDP were added 24 hours later , MTS assay was used to measure cell survival rate and cDDP on the cells of the half inhibitory concentration (IC50).4 μg/mL cDDP was added after 48 hours of interfering NC and S1+S2+S3.The apoptosis rate were detected by flow cytometry .Results:At the concentration of 0, 0.5, 1, 2, 4, 8 μg/mL cDDP, the survival rates of A549 cell with NC were 100% ±3.21%, 94.86% ±7.14%, 89.79% ±3.29%, 66.67% ±5.40%, 33.31% ± 1.79%, 19.12% ±0.41%, respectively;While survival rates of A549 cell with S1+S2+S3 were 100% ±6.53%,71.63% ±8.23%, 59.33%? ±0.94%,? 37.42%? ±1.57%, 19.41%? ±? 1.41%, 12.75% ±? 0.27%. The cell survival rate of NC and S1+S2+S3 group has significant difference at the same concentration of cDDP (P <0. 05).The IC50 of S1+S2+S3and NC were (1.25 ±0.11) and (3.09 ±0.36) μg/mL(P<0.01).The cell apoptosis rates of NC and S1+S2+S3 were 8.06%±1.79% and 14.96%±0.47% without cDDP(P<0.05), but the apoptosis rate were 24.61%±1.98% and 43.90%±3.01% with cDDP(P <0.05).Conclusion: Down-regulation ERCC1 by RNAi can enhance the effects of cDDP on promoting apoptosis and inhibiting proliferation of the A 549 cells.The sensitivity of A549 cell to cisplatin can be enhanced by RNA interfering ERCC 1 gene.
Keywords：ERCC1RNA interferencecDDPlung cancer
Publication Date：2014-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 4-7 )