The neutral red assay can be used to evaluate cell viability during autophagy or in an acidic microenvironment in vitro.
The neutral red assay can be used to evaluate cell viability during autophagy or in an acidic microenvironment in vitro.
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中性红色测定可用于评估自噬或体外酸性微环境中的细胞活力。
DOI：
10.1080/10520295.2020.1802065
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发表时间：
2021-05
期刊：
影响因子：
--
通讯作者：
McNally LR
中科院分区：
文献类型：
--
作者：
Gomez-Gutierrez JG;Bhutiani N;McNally MW;Chuong P;Yin W;Jones MA;Zeiderman MR;Grizzle WE;McNally LR
肿瘤微环境中的恶劣条件，如缺氧和细胞外酸性pH值（pHe），导致有限或无细胞毒性的恶性化疗。用于确定新开发药物效力的标准MTT、ATPlite和蛋白酶测定在缺氧或酸性条件下应用时经常给出错误的结果。因此，需要开发一种在缺氧和酸性pHe的情况下不产生假阳性或假阴性结果的细胞毒性测定。我们评估了目前使用的细胞活力测定以及中性红染色，以评估卡铂、吉西他滨或氯喹治疗后在酸性pH微环境中生长的卵巢癌和胰腺癌细胞的活力。我们使用蛋白质印迹法对胱天蛋白酶原-9和裂解的胱天蛋白酶-9以及LC 3-I和LC 3-II进行了细胞活力验证。标准细胞活力测定表明细胞活力准确地在pHe 7.4，但不相关的诱导细胞凋亡或自噬在酸性pH。相比之下，我们的改良中性红测定法在一定的pH范围内准确地检测细胞活力，这一点通过其与诱导凋亡和自噬的相关性来证明。中性红染色是有效的评价在酸性pH或缺氧条件下化疗剂对细胞活力的影响。
Harsh conditions within the tumor microenvironment, such as hypoxia and extracellular acidic pH (pHe), inactivate some chemotherapies, which results in limited or no cytotoxicity. Standard MTT, ATPlite and protease assays that are used to determine the potency of newly developed drugs often give erroneous results when applied under hypoxic or acidic conditions. Therefore, development of a cytotoxicity assay that does not yield false positive or false negative results under circumstances of both hypoxia and acidic pHe is needed. We evaluated currently used cell viability assays as well as neutral red staining to assess viability of ovarian and pancreatic cancer cells grown in an acidic pHe microenvironment after treatment with carboplatin, gemcitabine or chloroquine. We validated cell viability using western blotting of pro-caspase-9 and cleaved-caspase-9, and LC3-I and –II. Standard cell viability assays indicated cell viability accurately at pHe 7.4, but was not correlated with induction of apoptosis or autophagy at acidic pHe. By contrast, our modified neutral red assay detected cell viability accurately over a range of pHe as demonstrated by its correlation with induction of apoptosis and autophagy. Neutral red staining is effective for evaluating the effect of chemotherapeutic agents on cell viability under acidic pHe or hypoxic conditions.
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DOI：
10.1016/s1387-2656(05)11004-7
发表时间：
2005-01-01
期刊：
BIOTECHNOLOGY ANNUAL REVIEW, VOL 11
影响因子：
--
作者：
Berridge, MV;Herst, PM;Tan, AS
通讯作者：
Tan, AS
DOI：
10.1158/1078-0432.ccr-15-0314
发表时间：
2015-10-15
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Kimbrough CW;Khanal A;Zeiderman M;Khanal BR;Burton NC;McMasters KM;Vickers SM;Grizzle WE;McNally LR
通讯作者：
McNally LR
影响因子：
3
作者：
Johno, Hisashi;Takahashi, Shuhei;Kitamura, Masanori
通讯作者：
Kitamura, Masanori
影响因子：
5.7
作者：
Balic, Anamaria;Draeby Sorensen, Morten;Heeschen, Christopher
通讯作者：
Heeschen, Christopher
影响因子：
2.2
作者：
Egger, Michael E.;Huang, Justin S.;McNally, Lacey R.
通讯作者：
McNally, Lacey R.