中华消化病与影像杂志(电子版)
中華消化病與影像雜誌(電子版)
중화소화병여영상잡지(전자판)
2015年
3期
32-34
,共3页
姜琪琪%王莉%张红梅%王燕泽%季万胜
薑琪琪%王莉%張紅梅%王燕澤%季萬勝
강기기%왕리%장홍매%왕연택%계만성
肿瘤抑制蛋白质p53%氟尿嘧啶%胃肿瘤%细胞系,肿瘤
腫瘤抑製蛋白質p53%氟尿嘧啶%胃腫瘤%細胞繫,腫瘤
종류억제단백질p53%불뇨밀정%위종류%세포계,종류
Tumor suppressor protein p53%Fluorouracil%Stomach neoplasms%Cell line,tumor
目的:探讨p53异构体p53β在氟尿嘧啶(5-FU)干预胃癌细胞系MKN45及SGC-7901中表达的生物学效应。方法将不同浓度的5-FU作用于MKN45和SGC-7901细胞内24h,采用细胞增殖/毒性检测试剂盒( CCK-8试剂盒)检测细胞抑制率;巢式逆转录多聚酶链反应( nRT-PCR)检测p53和P53βmRNA的表达变化;Western-blot检测p53和P53β蛋白的表达变化。结果不同浓度(25、50、100μg/ml)的5-FU对MKN45细胞有抑制作用,抑制率分别为18.204%、31.354%、52.969%,随药物浓度的增加而增加;对SGC-7901细胞抑制率分别为26.305%、26.363%、25.780%,虽有抑制作用,但与药物浓度无关;浓度为100μg/ml 的5-FU作用24 h后,对MKN45细胞抑制率明显高于SGC-7901细胞。 nRT-PCR检测 MKN45中, p53和 P53βmRNA的表达随5-FU浓度的增加而增加。Western-blot检测MKN45中,p53和P53β蛋白的表达随5-FU浓度的增加而增加。结论5-FU对表达野生型p53的人胃癌细胞MKN45的抑制效应中,随5-FU浓度的增加,上调p53、p53β的表达,推测p53β在肿瘤生长抑制实验中发挥着重要的作用。
目的:探討p53異構體p53β在氟尿嘧啶(5-FU)榦預胃癌細胞繫MKN45及SGC-7901中錶達的生物學效應。方法將不同濃度的5-FU作用于MKN45和SGC-7901細胞內24h,採用細胞增殖/毒性檢測試劑盒( CCK-8試劑盒)檢測細胞抑製率;巢式逆轉錄多聚酶鏈反應( nRT-PCR)檢測p53和P53βmRNA的錶達變化;Western-blot檢測p53和P53β蛋白的錶達變化。結果不同濃度(25、50、100μg/ml)的5-FU對MKN45細胞有抑製作用,抑製率分彆為18.204%、31.354%、52.969%,隨藥物濃度的增加而增加;對SGC-7901細胞抑製率分彆為26.305%、26.363%、25.780%,雖有抑製作用,但與藥物濃度無關;濃度為100μg/ml 的5-FU作用24 h後,對MKN45細胞抑製率明顯高于SGC-7901細胞。 nRT-PCR檢測 MKN45中, p53和 P53βmRNA的錶達隨5-FU濃度的增加而增加。Western-blot檢測MKN45中,p53和P53β蛋白的錶達隨5-FU濃度的增加而增加。結論5-FU對錶達野生型p53的人胃癌細胞MKN45的抑製效應中,隨5-FU濃度的增加,上調p53、p53β的錶達,推測p53β在腫瘤生長抑製實驗中髮揮著重要的作用。
목적:탐토p53이구체p53β재불뇨밀정(5-FU)간예위암세포계MKN45급SGC-7901중표체적생물학효응。방법장불동농도적5-FU작용우MKN45화SGC-7901세포내24h,채용세포증식/독성검측시제합( CCK-8시제합)검측세포억제솔;소식역전록다취매련반응( nRT-PCR)검측p53화P53βmRNA적표체변화;Western-blot검측p53화P53β단백적표체변화。결과불동농도(25、50、100μg/ml)적5-FU대MKN45세포유억제작용,억제솔분별위18.204%、31.354%、52.969%,수약물농도적증가이증가;대SGC-7901세포억제솔분별위26.305%、26.363%、25.780%,수유억제작용,단여약물농도무관;농도위100μg/ml 적5-FU작용24 h후,대MKN45세포억제솔명현고우SGC-7901세포。 nRT-PCR검측 MKN45중, p53화 P53βmRNA적표체수5-FU농도적증가이증가。Western-blot검측MKN45중,p53화P53β단백적표체수5-FU농도적증가이증가。결론5-FU대표체야생형p53적인위암세포MKN45적억제효응중,수5-FU농도적증가,상조p53、p53β적표체,추측p53β재종류생장억제실험중발휘착중요적작용。
Objective To explore the expression of p53βand its biological effects in fluorouracil(5-FU) treated MKN45 and SGC-7901 gastric cancer cell lines.Methods MKN45 and SGC7901 cells were treated with 5-FU of different concentrations for 24 h.The growth inhibition rate was detected by CCK-8,and mRNA expressions of p53 and p53βwere measured by nested reverse transcription PCR ( nRT-PCR ) , and protein expressions of p53 and p53βwere measured by Western-blot.Results CCK-8 assay showed that the different concentrations of 5-FU(25,50 and 100μg/ml) had inhibiting effect on MNK45 cells,and the inhibition rates were 18.204%,31.354%,52.969%,which increased with the drug concentration increase. However,in SGC-7901 cells the inhibition rates were 26.305%,26.363%,25.780%, having statistically significance but not being associated with the drug concentration.After 24 h,the inhibition rate of 100μg/ml 5-FU in MKN45 was higher than that in SGS-7901.nRT-PCR and Western blot results showed that the expressions of p53 and p53βincreased with the increasing concentrations of 5-FU in MKN45 cells. Conclusions In the induction of 5-FU to the proliferation inhibition of MKN45 cells expressed wild? type p53,the up-regulation of p53βassociates with wide-type p53,which infers that p53βplays an important role in the inhibition effect of gastric cancer cell growth.