A new multiplex assay allowing simultaneous detection of the inhibition of cell proliferation and induction of cell death
A new multiplex assay allowing simultaneous detection of the inhibition of cell proliferation and induction of cell death
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DOI：
10.1002/jcb.20531
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发表时间：
2005-09-01
影响因子：
4
通讯作者：
Zerza-Schnitzhofer, G
中科院分区：
文献类型：
--
作者：
Wesierska-Gadek, J;Gueorguieva, M;Zerza-Schnitzhofer, G
在人类恶性肿瘤的化学疗法中使用的不同抗癌药物的功效在肿瘤组织之间变化，并且很大程度上取决于治疗剂同时抑制细胞增殖和通过细胞凋亡消除恶性细胞的能力。特别是，早期凋亡变化的检测似乎是重要的，因为细胞凋亡的早期阶段不同于那些坏死。因此，开发一种新的测试，允许快速和伴随筛选的抗增殖和促凋亡作用的抗癌药物的数量是非常感兴趣的。为此，我们选择了一个实验模型，顺铂（CP）对人宫颈癌HeLaS 3细胞的抗增殖和促凋亡作用。如前所述，暴露于CP的HeLaS 3导致伴随的细胞增殖抑制和诱导凋亡的剂量和时间依赖性的方式。在本研究中，我们进行了两个独立的方法。在第一种方法中，我们分别使用CellTiter-Glo（TM）发光细胞活力测定和Caspase-Glo（TM）3/7测定在两个单独的微量滴定板中检测细胞增殖和半胱天冬酶-3/7的活性。在第二种方法中，我们通过使用CellTiter-Blue（TM）细胞活力测定和Caspase-Glo（TM）3/7测定的多重测定在一个微量滴定板中顺序地测定相同的参数。这两种方法给出了非常相似的结果，表明这种新的多重测定为同时检测细胞数量和半胱天冬酶-3/7的活化提供了重要的优势。新的多重检测试剂盒提供了一系列优于标准检测试剂盒的优势。
The efficacy of distinct anti-cancer drugs used in the chemotherapy of human malignancies varies between tumor tissues and depends largely on the ability of the therapeutic agents to simultaneously inhibit cell proliferation and to eliminate malignant cells by apoptosis. Especially, detection of early apoptotic changes seems to be important because early stages of apoptosis differ from those of necrosis. Therefore, the development of a novel test allowing fast and concomitant screening of the anti-proliferative and pro-apoptotic action of a number of anti-cancer drugs is of great interest. For this purpose, we choose as an experimental model a well characterized anti-proliferative and pro-apoptotic effect of cisplatin (CP) on human cervical carcinoma HeLaS3 cells. As previously reported, exposure of HeLaS3 to CP resulted in a concomitant inhibition of cell proliferation and induction of apoptosis in a dose- and time-dependent manner. In the present study we performed two independent approaches. In the first approach, we examined the cell proliferation and activity of caspases-3/7 in two separate microtiter plates using the CellTiter-Glo (TM) Luminescent Cell Viability Assay and the Caspase-Glo (TM) 3/7 Assay, respectively. In the second approach, we determined the same parameters sequentially in one microtiter plate by a mutiplexing assay using CellTiter-Blue (TM) Cell Viability Assay and Caspase-Glo (TM) 3/7Assay. The both approaches gave very similar results indicating that this new multiplexing assay offers an important advantage for simultaneous detection of cell number and activation of caspases-3/7. The new multiplexing assay offers a range of benefits over standard assays.