VIBRATIONAL SPECTROSCOPY OF VIALBE; PAIRED TUMORIGENIC AND NON-TUMORIGENIC CELLS Page: 2 of 9
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Vibrational Spectroscopy of Viable, Paired Tumorigenic and Non-
Tumorigenic Cells
J. R. Mourant, Y. R. Yamada, S. Carpenter, A. Guerra, J. Schoonover, and J. P. Freyer
Los Alamos National Laboratory
Los Alamos, NM 87545
ABSTRACT
Infrared absorption of two pairs of non-tumorigenic and tumorigenic cells suspended in phosphate
buffered saline have been measured. Suspensions of cells with several different growth cycle
distributions were measured. The effect of both growth cycle and tumorigenity on the infrared
absorption spectrum will be presented. For example, changes in absorption in the phosphate
absorption region were observed for suspensions with different cell cycle distributions. We will
discuss the biochemistry which may cause these changes. We have found that spectra of isolated
nuclei allow the DNA spectra to be studied, without the confounding influence of RNA. Therefore,
the measurement of isolated nuclei may represent a method of detecting changes in DNA
architecture with cell cycle. As part of this exploratory study we have also examined the variation
in spectra with cell type and compared epithelial cells with fibroblast cells. Very little change was
observed. Similarly we saw very little change in the spectra of tumorigenic and non-tumorigenic
cells harvested with similar cell cycle distributions. Changes in the spectra were observed when
rapidly growing tumorigenic cells were compared to slowly replicating nontumorigenic cells.
INTRODUCTION
IR and Raman spectroscopic measurements can provide detailed molecular information.
Consequently, these spectroscopies have the potential to generate important biochemical
information for tissue diagnosis. Some of the medical applications of vibrational spectroscopy
currently under investigation include characterization of atherosclerosis (plaque in arteries), and
detection of cancer.
METHODS
Biochemical components of cells were obtained and measured in aqueous media. Type I "highly
polymerized" calf thymus DNA (Sigma) was dissolved in TE buffer (pH 8.02, 10mM TRIS, 1mM
EDTA). Type IV calf liver RNA was also dissolved in TE buffer. L-a-phosphotidylcholine and L-
x-phosphotidylethanolamine were dissolved in a buffer consisting of 0.01 M, pH 7.3 TRIS, 0.1 M
NaCl and 0.02 M sodium azide. Albumin was measured in a pH 7 phosphate buffer and lysozyme
was measured in pH 3.8 sodium citrate buffer.
Four types of cells derived from rat embryo fibroblast (REF) cells were used. M1 and Ratl cells are
immortalized but non-tumorigenic derivatives of REF cells. MR1 cells are a tumorigenic derivative
of M1 cells: specifically, these cells were transfected with the point-mutated T24Ha-ras-oncogene.
Similarly, Ratl-T1 cells are a tumorigenic derivative of Ratl cells obtained by a stable transfection
with the same mutant h-ras oncogene. Cells were cultured as monolayers in standard tissue culture
flasks using Dulbecco's modified eagle's medium containing 4.5 g/1 D-glucose, 5% (v/v) fetal calf
serum, 100 IU/ml penicillin, and 100 g/ml streptomycin.
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MOURANT, J. R.; YAMADA, Y. R. & AL, ET. VIBRATIONAL SPECTROSCOPY OF VIALBE; PAIRED TUMORIGENIC AND NON-TUMORIGENIC CELLS, article, December 1, 2001; New Mexico. (https://digital.library.unt.edu/ark:/67531/metadc719653/m1/2/: accessed April 25, 2024), University of North Texas Libraries, UNT Digital Library, https://digital.library.unt.edu; crediting UNT Libraries Government Documents Department.