姜琪琪,张 娜,郭 爱.rmhTNF对Δ133p53表达阳性胃癌细胞系MKN45的影响[J].中国肿瘤,2015,24(6):524-528.
rmhTNF对Δ133p53表达阳性胃癌细胞系MKN45的影响
Effect of rmhTNF on Human Gastric Cancer Cell Lines MKN45 with Δ133p53 Status
投稿时间:2014-10-07  
DOI:10.11735/j.issn.1004-0242.2015.06.A018
中文关键词:  rmhTNF  胃癌细胞系  Δ133p53  p53
英文关键词:rmhTNF  gastric cancer cell lines  Δ133p53  p53
基金项目:山东省优秀中青年科学家科研奖励基金 (BS2010SW034)
作者单位
姜琪琪 潍坊医学院 
张 娜 潍坊医学院 
郭 爱 潍坊医学院 
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中文摘要:
      摘 要:[目的] 利用rmhTNF-MKN45细胞模型,观察p53异构体Δ133p53及p53下游基因的表达。[方法] 不同浓度(50、100、200、400、500 IU/ml)重组改构人肿瘤坏死因子(rmhTNF)作用应用于MKN45,采用CCK-8法,观察其细胞增殖抑制率;通过巢式逆转录多聚酶链反应(nRT-PCR法)检测Δ133p53、p53、Gadd45α、PTEN、Mdm2、Bax和CyclinB1 mRNA的表达变化。[结果] 随rmhTNF作用浓度增加和作用时间(24、48、72h)延长,MKN45细胞抑制率明显增加(F=35.683,P<0.001;F=60.328,P<0.001)。随rmhTNF浓度增加,MKN45细胞中,p53、Gadd45α、PTEN、Bax基因表达上调,Δ133p53、Mdm2、CyclinB1基因表达下调。Δ133p53与p53、Gadd45α、PTEN、Bax表达呈负相关(r=-0.916,-0.894,-0.872,-0.971,P均<0.01),与Mdm2、CyclinB1表达呈正相关(r=0.924,0.943,P均<0.01)。[结论] 在肿瘤生长抑制效应中,Δ133p53异构体表现出与野生型p53拮抗作用,其机制可能与调控Gadd45α、PTEN、Mdm2、Bax 、CyclinB1等下游基因表达有关。
英文摘要:
      Abstract:[Purpose] To observe the expression and significant of the p53 isoforms Δ133p53 and p53 downstream molecules by rmhTNF-MKN45 gastric cancer cell line model. [Methods] The different concentrations (50、100、200、400、500 IU/ml) of recombined and modified human Tumor Necrosis Factor (rmhTNF) were used in gastric carcinoma cell lines MKN45. The growth inhibitory effect of rmhTNF was observed by CCK8 assay. The expression of p53 isoforms Δ133p53 and p53 downstream molecule Gadd45α,PTEN,Mdm2,Bax and CyclinB1 on the mRNA level was detected by nested reverse transcription PCR (nRT-PCR). [Results] With the increased concentration of rmhTNF and the role of the extension of time(24,48,72h),the cell inhibition rate increase gradually(F=35.683,P<0.001;F=60.328,P<0.001).In mRNA measurement,down-regulation of Δ133p53,Mdm2 and CyclinB1,up-regulation of p53,Gadd45α,PTEN and Bax was significant in MKN45 cells treated by rmhTNF. The expression of Δ133p53 was negatively related to p53,Gadd45α,PTEN and Bax (r=-0.916,-0.894,-0.872,-0.971;all P<0.01),but positively related to Mdm2 and CyclinB1 (r=0.924,=0.943;all P<0.01).[Conclusion] Δ133p53 is shown as an anti-p53 factor,which might be related to regulate Gadd45α,PTEN,Mdm2,Bax and CyclinB1 in the inhibitory effects of rmhTNF to gastric carcinoma.
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