[KSCI] Korea Science Citation Index Service

http://dx.doi.org/10.4333/KPS.2006.36.5.309

Simultaneous Evaluation of Cellular Vitality and Drug Penetration in Multicellular Layers of Human Cancer Cells

Al-Abd Ahmed Mohammed (Department of Biomedical Sciences, College of Medicine, The Catholic University of Korea)
Lee Joo-Ho (Department of Biomedical Sciences, College of Medicine, The Catholic University of Korea)
Kuh Hyo-Jeong (Department of Biomedical Sciences, College of Medicine, The Catholic University of Korea)

Publication Information

Journal of Pharmaceutical Investigation / v.36, no.5, 2006 , pp. 309-314 More about this Journal

Abstract

The multicellular layers(MCL) of human cancer cells is a three dimensional(3D) in vitro model for human solid tumors which has been used primarily for the assessment of avascular penetration of anti-cancer drugs. For anti-cancer drugs with penetration problem, MCL represents a good experimental model that can provide clinically relevant data. Calcein-AM is a fluorescent dye that demonstrates the cellular vitality in a graded manner in cancer cell culture system. In the present study, we evaluated the use of calcein-AM for determination of anti-proliferative activity of anti-cancer agents in MCL model of DLD-1 human colorectal cancer cells. Optical sectioning of confocal imaging was compromised with photonic attenuation and penetration barrier in the deep layers of MCL. By contrast, fluorescent measurement on the cryo-sections provided a feasible alternative. Cold pre-incubation did not enhance the calcein-AM distribution to a significant degree in MCL of DLD-1 cells. However, the simultaneous determination of drug penetration and cellular vitality appeared to be possible in drug treated MCL. In conclusion, these data suggest that calcein-AM can be used for the simultaneous determination of drug-induced anti-proliferative effect and drug penetration in MCL model.

Keywords

Multicellular layers(MCL); Solid tumor; Penetration; Calcein-AM; Confocal imaging;

Citations & Related Records

Reference

1	R.C. Bates, N.S. Edwards and J.D. Yates, Spheroids and cell survival. Crit. Rev. Oncol. Hematol, 36, 61-74 (2000) DOI ScienceOn
2	B.C. Baguley and E.S. Marshall, In vitro modelling of human tumour behaviour in drug discovery programmes. Eur. J. Cancer, 40, 794-801 (2004) DOI ScienceOn
3	K.O. Hicks, F.B. Pruijn, J.R. Sturman, W.A. Denny and W.R. Wilson, Multicellular resistance to tirapazamine is due to restricted extravascular transport: A pharmacokinetic/pharma- codynamic study in HT29 multicellular layer cultures. Cancer Res., 63, 5970-7 (2003)
4	M. Wartenberg, J. Hescheler, H. Acker, H. Diedershagen and H. Sauer, Doxorubicin distribution in multicellular prostate cancer spheroids evaluated by confocal laser scanning microscopy and the 'optical probe technique'. Cytometry, 31, 137-45 (1998) DOI ScienceOn
5	C.M. Lee and I.F. Tannock, Inhibition of endosomal seque- stration of basic anticancer drugs: Influence on cytotoxicity and tissue penetration. Br J. Cancer, 94, 863-9 (2006) DOI ScienceOn
6	K. Yazawa, N.H. Tsuno, J. Kitayama, K. Kawai, Y. Okaji, M. Asakage, E. Sunami, S. Kaisaki, N. Hori, T. Watanabe, K. Takahashi and H. Nagawa, Selective inhibition of cyclooxy- genase-2 inhibits colon cancer cell adhesion to extracellular matrix by decreased expression of beta1 integrin. Cancer Sci., 96, 93-9 (2005) DOI ScienceOn
7	A.A. Neyfakh, Use of fluorescent dyes as molecular probes for the study of multidrug resistance. Exp. Cell Res., 174, 168-76 (1988) DOI ScienceOn
8	C. Martin, J. Walker, A. Rothnie and R. Callaghan, The expression of P-glycoprotein does influence the distribution of novel fluorescent compounds in solid tumour models. Br J. Cancer, 89, 1581-9 (2003) DOI ScienceOn
9	T. Nederman and P. Twentyman, Spheroids for studies of drug effects. Recent Results Cancer Res, 95, 84-102 (1984)
10	E. Smitskamp-Wilms, H.M. Pinedo, G. Veerman, V.W. Ruiz van Haperen and G.J. Peters, Postconfluent multilayered cell line cultures for selective screening of gemcitabine. Eur. J. Cancer, 34, 921-6 (1998) DOI ScienceOn
11	S.H. Jang, M.G. Wientjes and J.L. Au, Determinants of paclitaxel uptake, accumulation and retention in solid tumors. Invest New Drugs, 19, 113-23 (2001) DOI ScienceOn
12	J.M. Padron and G.J. Peters, Cytotoxicity of sphingoid marine compound analogs in mono- and multilayered solid tumor cell cultures. Invest New Drugs, 24, 195-202 (2006) DOI
13	M.T. Santini, G. Rainaldi and P.L. Indovina, Apoptosis, cell adhesion and the extracellular matrix in the three-dimensional growth of multicellular tumor spheroids. Crit. Rev. Oncol. Hematol, 36, 75-87 (2000) DOI ScienceOn
14	M. Wartenberg and H. Acker, Quantitative recording of vitality patterns in living multicellular spheroids by confocal microscopy. Micron, 26, 395-404 (1995) DOI ScienceOn
15	B. Desoize, Contribution of three-dimensional culture to cancer research. Crit. Rev. Oncol. Hematol, 36, 59-60 (2000) DOI ScienceOn
16	R. Grantab, S. Sivananthan and I.F. Tannock, The penetration of anticancer drugs through tumor tissue as a function of cellular adhesion and packing density of tumor cells. Cancer Res., 66, 1033-9 (2006) DOI ScienceOn
17	A.H. Kyle, L.A. Huxham, A.S. Chiam, D.H. Sim and A.I. Minchinton, Direct assessment of drug penetration into tissue using a novel application of three-dimensional cell culture. Cancer Res., 64, 6304-9 (2004) DOI ScienceOn
18	J.K. Tunggal, T. Melo, J.R. Ballinger and I.F. Tannock, The influence of expression of P-glycoprotein on the penetration of anticancer drugs through multicellular layers. Int. J. Cancer, 86, 101-7 (2000) DOI ScienceOn
19	A.I. Minchinton and I.F. Tannock, Drug penetration in solid tumours. Nat. Rev. Cancer, 6, 583-92 (2006) DOI ScienceOn
20	R.M. Phillips, P.M. Loadman and B.P. Cronin, Evaluation of a novel in vitro assay for assessing drug penetration into avascular regions of tumours. Br J. Cancer, 77, 2112-9 (1998) DOI ScienceOn
21	D.S. Cowan, K.O. Hicks and W.R. Wilson, Multicellular membranes as an in vitro model for extravascular diffusion in tumours. Br J. Cancer Suppl, 27, S28-31 (1996)