Comparison of caspase-3 activation in tumor cells upon treatment of chemotherapeutic drugs using capillary electrophoresis

doi:10.1007/s13238-012-2008-7

Prot Cell

2012, Vol. 3

Issue (5) : 392-399 https://doi.org/10.1007/s13238-012-2008-7 PMID: 22528748

RESEARCH ARTICLE

Comparison of caspase-3 activation in tumor cells upon treatment of chemotherapeutic drugs using capillary electrophoresis

Shuang Sha^1,2, Honglin Jin^1,2, Xiao Li^1,2, Jie Yang^1,2, Ruiting Ai³, Jinling Lu^1,2(

)

1. Britton Chance Center for Biomedical Photonics-Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan 430074, China; 2. Key Laboratory of Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology, Wuhan 430074, China; 3. China National Center for Biotechnology Development, Beijing 100036, China

Download: PDF(486 KB) HTML
Export: BibTeX | EndNote | Reference Manager | ProCite | RefWorks

Abstract

Caspases play important roles in cell apoptosis. Measurement of the dynamics of caspase activation in tumor cells not only facilitates understanding of the molecular mechanisms of apoptosis but also contributes to the development, screening, and evaluation of anticancer drugs that target apoptotic pathways. The fluorescence resonance energy transfer (FRET) technique provides a valuable approach for defining the dynamics of apoptosis with high spatio-temporal resolution. However, FRET generally functions in the single-cell level and becomes ineffective when applied in the high throughput detection of caspase activation. In the current study, a FRET sensor was combined with capillary electrophoresis (CE) to achieve a high throughput method for cellular caspase detection. The FRET-based CE system is composed of a homemade CE system and a laser source for detecting the dynamics of caspase-3 in various cells expressing sensors of caspase-3 that have been treated with anticancer drugs, such as cell cycle-independent drug cisplatin and specific cell cycle drugs camptothecin and etoposide, as well as their combination with tumor necrosis factor (TNF). A positive correlation between the caspase-3 activation velocity and drug concentration was observed when the cells were treated with cisplatin, but cells induced by camptothecin and etoposide did not show any apparent correlation with their concentrations. Moreover, different types of cells presented distinct sensitivities under the same drug treatment, and the combination treatment of TNF and anticancer drugs significantly accelerated the caspase-3 activation process. Its high throughput capability and detection sensitivity make the FRET-based CE system a useful tool for investigating the mechanisms of anticancer drugs and anticancer drug screening.

Keywords apoptosis caspase-3 fluorescence resonance energy transfer (FRET) capillary electrophoresis (CE)

Corresponding Author(s): Lu Jinling,Email:lujinling@mail.hust.edu.cn

Issue Date: 01 May 2012

Cite this article:

Honglin Jin,Xiao Li,Jie Yang, et al. Comparison of caspase-3 activation in tumor cells upon treatment of chemotherapeutic drugs using capillary electrophoresis[J]. Prot Cell, 2012, 3(5): 392-399.

URL:

https://academic.hep.com.cn/pac/EN/10.1007/s13238-012-2008-7
https://academic.hep.com.cn/pac/EN/Y2012/V3/I5/392

1	Abbott, B.P., Abbott, R., Acernese, F., Adhikari, R., Ajith, P., Allen, B., Allen, G., Alshourbagy, M., Amin, R.S., Anderson, S.B., ., and the LIGO Scientific Collaboration & Virgo Collaboration. (2009). An upper limit on the stochastic gravitational-wave background of cosmological origin. Nature 460, 990-994 . doi: 10.1038/nature08278
2	Berezovski, M., Li, W.P., Poulter, C.D., and Krylov, S.N. (2002). Measuring the activity of farnesyltransferase by capillary electrophoresis with laser-induced fluorescence detection. Electrophoresis 23, 3398-3403 . doi: 10.1002/1522-2683(200210)23:19<3398::AID-ELPS3398>3.0.CO;2-Y
3	Bergeron, S., Beauchemin, M., and Bertrand, R. (2004). Camptothecin- and etoposide-induced apoptosis in human leukemia cells is independent of cell death receptor-3 and-4 aggregation but accelerates tumor necrosis factor-related apoptosis-inducing ligand-mediated cell death. Mol Cancer Ther 3, 1659-1669 .
4	Burton, E.R., and Libutti, S.K. (2009). Targeting TNF-alpha for cancer therapy. J Biol 8, 85. doi: 10.1186/jbiol189
5	Chen, A.K., and Tsourka, A. (2009). Imaging RNA in living cells with molecular beacons: current perspectives and challenges. J Innovative Opt Health Sci 2, 315-324 . doi: 10.1142/S179354580900070X
6	Chen, A.Y., and Liu, L.F. (1994). DNA topoisomerases: essential enzymes and lethal targets. Annu Rev Pharmacol Toxicol 34, 191-218 . doi: 10.1146/annurev.pa.34.040194.001203
7	de Jonge, M.J., Kaye, S., Verweij, J., Brock, C., Reade, S., Scurr, M., van Doorn, L., Verheij, C., Loos, W., Brindley, C., . (2004). Phase I and pharmacokinetic study of XR11576, an oral topoisomerase I and II inhibitor, administered on days 1-5 of a 3-weekly cycle in patients with advanced solid tumours. Br J Cancer 91, 1459-1465 .
8	Hara, K., Okamoto, M., Aki, T., Yagita, H., Tanaka, H., Mizukami, Y., Nakamura, H., Tomoda, A., Hamasaki, N., and Kang, D. (2005). Synergistic enhancement of TRAIL- and tumor necrosis factor alpha-induced cell death by a phenoxazine derivative. Mol Cancer Ther 4, 1121-1127 . doi: 10.1158/1535-7163.MCT-05-0067
9	Hsu, J.L., Chiang, P.C., and Guh, J.H. (2009). Tunicamycin induces resistance to camptothecin and etoposide in human hepatocellular carcinoma cells: role of cell-cycle arrest and GRP78. Naunyn Schmiedebergs Arch Pharmacol 380, 373-382 . doi: 10.1007/s00210-009-0453-5
10	Johnstone, R.W., Ruefli, A.A., and Lowe, S.W. (2002). Apoptosis: a link between cancer genetics and chemotherapy. Cell 108, 153-164 . doi: 10.1016/S0092-8674(02)00625-6
11	Jones, J., Heim, R., Hare, E., Stack, J., and Pollok, B.A. (2000). Development and application of a GFP-FRET intracellular cas pase assay for drug screening. J Biomol Screen 5, 307-318 . doi: 10.1177/108705710000500502
12	Joseph, J., Seervi, M., Sobhan, P.K., and Retnabai, S.T. (2011). High throughput ratio imaging to profile caspase activity: potential application in multiparameter high content apoptosis analysis and drug screening. PLoS One 6, e20114. doi: 10.1371/journal.pone.0020114
13	Lin, J., Zhang, Z., Zeng, S., Zhou, S., Liu, B.F., Liu, Q., Yang, J., and Luo, Q. (2006). TRAIL-induced apoptosis proceeding from caspase-3-dependent and-independent pathways in distinct HeLa cells. Biochem Biophys Res Commun 346, 1136-1141 . doi: 10.1016/j.bbrc.2006.05.209
14	Locke, S., and Figeys, D. (2000). Techniques for the optimization of proteomic strategies based on head column stacking capillary electrophoresis. Anal Chem 72, 2684-2689 . doi: 10.1021/ac0003293
15	Luo, K.Q., Yu, V.C., Pu, Y., and Chang, D.C. (2003). Measuring dynamics of caspase-8 activation in a single living HeLa cell during TNFalpha-induced apoptosis. Biochem Biophys Res Commun 304, 217-222 . doi: 10.1016/S0006-291X(03)00559-X
16	Mahajan, N.P., Harrison-Shostak, D.C., Michaux, J., and Herman, B. (1999). Novel mutant green fluorescent protein protease substrates reveal the activation of specific caspases during apoptosis. Chem Biol 6, 401-409 . doi: 10.1016/S1074-5521(99)80051-9
17	Namura, S., Zhu, J., Fink, K., Endres, M., Srinivasan, A., Tomaselli, K.J., Yuan, J., and Moskowitz, M.A. (1998). Activation and cleavage of caspase-3 in apoptosis induced by experimental cerebral ischemia. J Neurosci 18, 3659-3668 .
18	Neefjes, J., and Dantuma, N.P. (2004). Fluorescent probes for proteolysis: tools for drug discovery. Nat Rev Drug Discov 3, 58-69 . doi: 10.1038/nrd1282
19	Postiglione, I., Chiaviello, A., and Palumbo, G. (2011). Enhancing Photodynamyc Therapy Efficacy by Combination Therapy: Dated, Current and Oncoming Strategies. Cancers 3, 2597-2629 . doi: 10.3390/cancers3022597
20	Reinhold, W.C., Kouros-Mehr, H., Kohn, K.W., Maunakea, A.K., Lababidi, S., Roschke, A., Stover, K., Alexander, J., Pantazis, P., Miller, L., . (2003). Apoptotic susceptibility of cancer cells selected for camptothecin resistance: gene expression profiling, functional analysis, and molecular interaction mapping. Cancer Res 63, 1000-1011 .
21	Szlosarek, P.W., and Balkwill, F.R. (2003). Tumour necrosis factor alpha: a potential target for the therapy of solid tumours. Lancet Oncol 4, 565-573 . doi: 10.1016/S1470-2045(03)01196-3
22	Tsuruo, T., Naito, M., Tomida, A., Fujita, N., Mashima, T., Sakamoto, H., and Haga, N. (2003). Molecular targeting therapy of cancer: drug resistance, apoptosis and survival signal. Cancer Sci 94, 15-21 . doi: 10.1111/j.1349-7006.2003.tb01345.x
23	Tyas, L., Brophy, V.A., Pope, A., Rivett, A.J., and Tavaré, J.M. (2000). Rapid caspase-3 activation during apoptosis revealed using fluorescence-resonance energy transfer. EMBO Rep 1, 266-270 . doi: 10.1093/embo-reports/kvd050
24	Wu, Y.X., Xing, D., Luo, S.M., Tang, Y.H., and Chen, Q. (2006). Detection of caspase-3 activation in single cells by fluorescence resonance energy transfer during photodynamic therapy induced apoptosis. Cancer Lett 235, 239-247 . doi: 10.1016/j.canlet.2005.04.036
25	Yang, J., Zhang, Z., Lin, J., Lu, J., Liu, B.F., Zeng, S., and Luo, Q. (2007). Detection of MMP activity in living cells by a genetically encoded surface-displayed FRET sensor. Biochim Biophys Acta 1773, 400-407 . doi: 10.1016/j.bbamcr.2006.11.002
26	Yao, X., Panichpisal, K., Kurtzman, N., and Nugent, K. (2007). Cisplatin nephrotoxicity: a review. Am J Med Sci 334, 115-124 . doi: 10.1097/MAJ.0b013e31812dfe1e
27	Zarrine-Afsar, A., and Krylov, S.N. (2003). Use of capillary electrophoresis and endogenous fluorescent substrate to monitor intracellular activation of protein kinase A. Anal Chem 75, 3720-3724 . doi: 10.1021/ac034463+
28	Zhang, Z., Lin, J., Chu, J., Ma, Y., Zeng, S., and Luo, Q. (2008a). Activation of caspase-3 noninvolved in the bystander effect of the herpes simplex virus thymidine kinase gene/ganciclovir (HSV-tk/GCV) system. J Biomed Opt 13, 031209. doi: 10.1117/1.2937830
29	Zhang, Z., Yang, J., Lu, J., Lin, J., Zeng, S., and Luo, Q. (2008b). Fluorescence imaging to assess the matrix metalloproteinase activity and its inhibitor in vivo. J Biomed Opt 13, 011006. doi: 10.1117/1.2830659
30	Zhou, S., Lin, J., Du, W., Zhang, Z., Luo, Q., Liu, B.F., and Dai, Y. (2006a). Characterization of proteinase activation dynamics by capillary electrophoresis conjugating with fluorescent protein-based probe. J Chromatogr B Analyt Technol Biomed Life Sci 844, 158-162 . doi: 10.1016/j.jchromb.2006.06.038
31	Zhou, S.X., Lin, J.Q., Du, W., Zhang, Z.H., Luo, Q.M., Liu, B.F., and Dai, Y.Q. (2006b). Monitoring of proteinase activation in cell apoptosis by capillary electrophoresis with bioengineered fluorescent probe. Anal Chim Acta 569, 176-181 . doi: 10.1016/j.aca.2006.03.088
32	Zhu, L., Liu, T.C.-Y., Wu, M., Yuan, J.-Q., and Chen, T.-S. (2009). Extraocular Cellular Phototransduction. J Innovative Opt Health Sci 2, 93-100 . doi: 10.1142/S1793545809000358
33	Zuryn, A., Grzanka, A., Stepien, A., Grzanka, D., Debski, R., and Smolinski, D. (2007). Expression of cyclin A in human leukemia cell line HL-60 following treatment with doxorubicin and etoposide: the potential involvement of cyclin A in apoptosis. Oncol Rep 17, 1013-1019 .

[1]	Qiang Hong, Cong Li, Ruhong Ying, Heming Lin, Jingqiu Li, Yu Zhao, Hanhua Cheng, Rongjia Zhou. Loss-of-function of sox3 causes follicle development retardation and reduces fecundity in zebrafish[J]. Protein Cell, 2019, 10(5): 347-364.
[2]	Yuanlong Ge, Shu Wu, Zepeng Zhang, Xiaocui Li, Feng Li, Siyu Yan, Haiying Liu, Junjiu Huang, Yong Zhao. Inhibition of p53 and/or AKT as a new therapeutic approach specifically targeting ALT cancers[J]. Protein Cell, 2019, 10(11): 808-824.
[3]	Ping Wang, Zunpeng Liu, Xiaoqian Zhang, Jingyi Li, Liang Sun, Zhenyu Ju, Jian Li, Piu Chan, Guang-Hui Liu, Weiqi Zhang, Moshi Song, Jing Qu. CRISPR/Cas9-mediated gene knockout reveals a guardian role of NF-κB/RelA in maintaining the homeostasis of human vascular cells[J]. Protein Cell, 2018, 9(11): 945-965.
[4]	Haiyang Zhang,Jingjing Duan,Yanjun Qu,Ting Deng,Rui Liu,Le Zhang,Ming Bai,Jialu Li,Tao Ning,Shaohua Ge,Xia Wang,Zhenzhen Wang,Qian Fan,Hongli Li,Guoguang Ying,Dingzhi Huang,Yi Ba. Onco-miR-24 regulates cell growth and apoptosis by targeting BCL2L11 in gastric cancer[J]. Protein Cell, 2016, 7(2): 141-151.
[5]	Qian Fan,Xiangrui Meng,Hongwei Liang,Huilai Zhang,Xianming Liu,Lanfang Li,Wei Li,Wu Sun,Haiyang Zhang,Ke Zen,Chen-Yu Zhang,Zhen Zhou,Xi Chen,Yi Ba. miR-10a inhibits cell proliferation and promotes cell apoptosis by targeting BCL6 in diffuse large B-cell lymphoma[J]. Protein Cell, 2016, 7(12): 899-912.
[6]	Chao Lu,Yang Yang,Ran Zhao,Bingxuan Hua,Chen Xu,Zuoqin Yan,Ning Sun,Ruizhe Qian. Role of circadian gene Clock during differentiation of mouse pluripotent stem cells[J]. Protein Cell, 2016, 7(11): 820-832.
[7]	Fan Chen,Jiebo Chen,Jiacheng Lin,Anton V. Cheltsov,Lin Xu,Ya Chen,Zhiping Zeng,Liqun Chen,Mingfeng Huang,Mengjie Hu,Xiaohong Ye,Yuqi Zhou,Guanghui Wang,Ying Su,Long Zhang,Fangfang Zhou,Xiao-kun Zhang,Hu Zhou. NSC-640358 acts as RXRα ligand to promote TNFα-mediated apoptosis of cancer cell[J]. Protein Cell, 2015, 6(9): 654-666.
[8]	Xiangxuan Zhao,Yong Liu,Lei Du,Leya He,Biyun Ni,Junbo Hu,Dahai Zhu,Quan Chen. Threonine 32 (Thr32) of FoxO3 is critical for TGF-β-induced apoptosis via Bim in hepatocarcinoma cells[J]. Protein Cell, 2015, 6(2): 127-138.
[9]	Anna Gortat,Mónica Sancho,Laura Mondragón,Àgel Messeguer,Enrique Pérez-Payá,Mar Orzáez. Apaf1 inhibition promotes cell recovery from apoptosis[J]. Protein Cell, 2015, 6(11): 833-843.
[10]	Youguang Luo,Dengwen Li,Jie Ran,Bing Yan,Jie Chen,Xin Dong,Zhu Liu,Ruming Liu,Jun Zhou,Min Liu. End-binding protein 1 stimulates paclitaxel sensitivity in breast cancer by promoting its actions toward microtubule assembly and stability[J]. Protein Cell, 2014, 5(6): 469-479.
[11]	Xiao-Xi Guo,Yang Li,Chao Sun,Dan Jiang,Ying-Jia Lin,Feng-Xie Jin,Seung-Ki Lee,Ying-Hua Jin. p53-dependent Fas expression is critical for Ginsenoside Rh2 triggered caspase-8 activation in HeLa cells[J]. Protein Cell, 2014, 5(3): 224-234.
[12]	Guanghua Xu,Jing Wang,George Fu Gao,Cui Hua Liu. *Insights into battles between Mycobacterium tuberculosis* and macrophages**[J]. Protein Cell, 2014, 5(10): 728-736.
[13]	Yi Sun, Hua Li. Functional characterization of SAG/RBX2/ROC2/RNF7, an antioxidant protein and an E3 ubiquitin ligase[J]. Prot Cell, 2013, 4(2): 103-116.
[14]	Yide Mei, Mian Wu. Multifaceted functions of Siva-1: more than an Indian God of Destruction[J]. Prot Cell, 2012, 3(2): 117-122.
[15]	Xiaoping Zhang, Changjiang Weng, Yuan Li, Xiaoyan Wang, Chunsun Jiang, Xuemei Li, Youli Xu, Quan Chen, Lei Pan, Hong Tang. Human Bop is a novel BH3-only member of the Bcl-2 protein family[J]. Prot Cell, 2012, 3(10): 790-801.

Viewed

Full text

Abstract

Cited

Shared

Discussed