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[应用逆转录聚合酶链反应检测顺铂耐药结肠癌细胞中的耐药基因。I]

[Detection of drug resistant-gene in cisplatin-resistant colon carcinoma cells by RT-PCR assay. I].

作者信息

Funato T, Tone T, Miyachi H, Ishimori A

机构信息

Department of Clinical and Laboratory Medicine, Tohoku University School of Medicine, Sendai.

出版信息

Rinsho Byori. 1993 Jan;41(1):95-100.

PMID:8355415
Abstract

Expression of the TS and DNA polymerase beta genes as a drug resistant-gene were compared with the human colon carcinoma cell line HCT8S and a subline that was 4.5 fold resistant to cisplatin. Resistant cells (HCT8DDP) exhibited 3.4 fold and 2.5 fold increase in mRNA for both TS and DNA polymerase beta gene when compared with the parent cells by Northern blotting analysis, respectively. The RT (reverse transcription)-PCR (polymerase chain reaction) method has been modified to quantify a sequence of a drug resistance gene. This method exhibited greater sensitivity than conventional methods (Northern blotting analysis), requiring less than 1 fetogram of mRNA from cell lines. The RT-PCR assay could be an effective device in the early detection of resistance to chemotherapy.

摘要

将胸苷合成酶(TS)和DNA聚合酶β基因作为耐药基因的表达,与人类结肠癌细胞系HCT8S及对顺铂耐药4.5倍的一个亚系进行了比较。通过Northern印迹分析,与亲本细胞相比,耐药细胞(HCT8DDP)中TS和DNA聚合酶β基因的mRNA分别增加了3.4倍和2.5倍。逆转录(RT)-聚合酶链反应(PCR)方法已被改进以定量耐药基因的序列。该方法比传统方法(Northern印迹分析)具有更高的灵敏度,所需细胞系mRNA少于1飞克。RT-PCR检测可能是化疗耐药早期检测的一种有效手段。

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