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使用乳酸脱氢酶对新鲜和连续肿瘤细胞培养物进行化学敏感性测试。

Chemosensitivity testing of fresh and continuous tumor cell cultures using lactate dehydrogenase.

作者信息

Coley H M, Lewandowicz G, Sargent J M, Verrill M W

机构信息

CRC Centre for Cancer Therapeutics, Institute of Cancer Research, Sutton, Surrey, U.K.

出版信息

Anticancer Res. 1997 Jan-Feb;17(1A):231-6.

PMID:9066657
Abstract

We have examined the use of the LDH (lactate dehydrogenase) assay for chemosensitivity testing in established and primary cultures of sarcoma, leukaemia and ovarian cancer in parallel with the MTT assay. The method we describe is rapid, sensitive and ideal for 96-well plate assays using adherent or suspension cultures. Excellent agreement between the two methods was observed (r = 0.936) using a variety of antitumour agents, with some notable exceptions. In the Bax (human synovial sarcoma) cell line MTT colour production by control cells was very low, thus MTT-->formazan production could not be relied upon as a definitive end point equating with cell number. In contrast, colour production of control cells using the LDH assay was significantly greater and all cultures tested were suitable for titration of chemosensitivity. There was a discrepancy between IC50 values obtained either by cell counting or MTT in the HTB88 (human leiomyosarcoma) line treated with 5-FU (59.9 microM vs > 200 microM, respectively). However, cell counting agreed well with the LDH assay (IC50 47.3 microM). Whilst the MTT assay remains a reliable method for chemosensitivity testing, the LDH assay may prove more appropriate in certain experimental settings.

摘要

我们已将乳酸脱氢酶(LDH)检测法与MTT检测法并行用于肉瘤、白血病和卵巢癌的传代培养及原代培养的化学敏感性检测。我们所描述的方法快速、灵敏,适用于使用贴壁或悬浮培养物的96孔板检测。使用多种抗肿瘤药物时,观察到两种方法之间具有极佳的一致性(r = 0.936),但也有一些明显的例外情况。在Bax(人滑膜肉瘤)细胞系中,对照细胞产生的MTT颜色非常低,因此不能将MTT转化为甲臜产物作为等同于细胞数量的确定终点。相比之下,使用LDH检测法时对照细胞产生的颜色明显更深,并且所有测试的培养物都适合化学敏感性滴定。在用5-氟尿嘧啶(5-FU)处理的HTB88(人平滑肌肉瘤)细胞系中,通过细胞计数或MTT获得的IC50值之间存在差异(分别为59.9 microM和> 200 microM)。然而,细胞计数与LDH检测法结果吻合良好(IC50为47.3 microM)。虽然MTT检测法仍然是化学敏感性检测的可靠方法,但在某些实验环境中,LDH检测法可能更合适。

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