ISSN 16076729, Doklady Biochemistry and Biophysics, 2011, Vol. 437, pp. 105–108. © Pleiades Publishing, Ltd., 2011.
Original Russian Text © K.N. Kashkin, E.A. Musatkina, A.V. Komelkov, D.A. Sakharov, E.V. Trushkin, E.A. Tonevitsky, T.V. Vinogradova, E.P. Kopantzev, M.V. Zinovyeva,
O.V.Kovaleva, K.A. Arkhipova, I.B. Zborovskaya, A.G. Tonevitsky, E.D. Sverdlov, 2011, published in Doklady Akademii Nauk, 2011, Vol. 437, No. 6, pp. 837–841.
105
Paclitaxel is a natural substance from the group of
taxanes and one of the most widely used drugs in che
motherapy of malignant tumors, including the non
small cell lung cancer (NSCLC). Treatment of
patients with advanced NSCLC with paclitaxel alone
or in combination with other drugs led to complete or
partial tumor regression in at most 41% of patients [1].
Individual selection of drugs that are effective for a
particular patient is one of the hottest approaches in
the strategy to improve the results of chemotherapeu
tic treatment of cancer patients. This work is dedicated
to finding new markers and studying the resistance
mechanisms of lung cancer to paclitaxel.
The main mechanism of action of paclitaxel is the
blockade of cell division due to its specific binding to
β3 tubulin and stabilization of microtubules. An
increased expression of some βtubulin isotypes corre
lates with resistance to paclitaxel and the degree of
malignancy of tumors of the lung, prostate, ovary and
other tissues. The same result can be caused by muta
tions of βtubulins, although the clinical significance
of mutations is controversial (see reviews [2, 3]).
It is known that the resistance of tumor cells to
paclitaxel may be associated with an increased expres
sion or polymorphism of some ABCtransporters
responsible for multidrug resistance [4] as well as with
activity of growth factor receptors (EGFR and HER
2), proteins LIMK1, LIMK2, TGFB1, STMN1, spin
dle checkpoint proteins, apoptosis regulators, and
others [2, 3].
The use of DNA microchips [5–10] made it possi
ble to reveal the sets of genes whose expression corre
lates with chemoresistance of lung cancer cells. How
ever, the sets of genes that, with a certain probability,
may be involved in resistance development, which
were proposed by different authors, differ. In addition,
the genes detected in cell cultures may be uninforma
tive for predicting the response of patients to chemo
therapy [11]. At the same time, studies of clinical
specimens only may give ambiguous results due to
polymorphisms, which lead to a high interindividual
variability of systems involved in the development of
chemoresistance of cancer patients, as well as because
of chemotherapy regimens that usually include several
drugs.
To search for new informative markers and to reveal
the molecular mechanisms of drug resistance in
tumors, we studied the relationship between the sensi
tivity of lung cancer cells to paclitaxel and the expres
sion of a wide range of genes using the new microchip
platform Affymetrix Human GeneChip ST1.0, which
contained probes for over 28000 human mRNAs.
As a biological model, we used lung cancer cells of
six lines derived from ATCC (A549, NCIH292, NCI
H460, and NCIH1299) and ECACC (NCIH322
and NCIH358). Although the sensitivity of certain
lines to various drugs is known (http://dis
cover.nci.nih.gov/cellminer/), we redefined the IC50
of paclitaxel for all cells using the MTT technique [12]
in order to eliminate the effect of subculturing of cells
on their sensitivity to the drug. For each cell line, we
analyzed the results of at least three significant mea
surements (Fig. 1).
Hybridization of labeled samples prepared from
total cell RNA with microchips was performed using
the equipment and method of Affymetrix. The results
of hybridization were processed by the RMA algo
rithm using the xps library (Christian Stratowa;
http://www.bioconductor.org) in R system. Fluores
cence signals were filtered using I/INI algorithm [13];
signals with random variation were ignored.
Genes Potentially Associated with Resistance
of Lung Cancer Cells to Paclitaxel
K. N. Kashkin
a
, E. A. Musatkina
b
, A. V. Komelkov
b
, D. A. Sakharov
c
, E. V. Trushkin
c
,
E. A. Tonevitsky
c
, T. V. Vinogradova
a
, E. P. Kopantzev
a
, M. V. Zinovyeva
a
, O. V. Kovaleva
b
,
K. A. Arkhipova
b
, I. B. Zborovskaya
b
, Corresponding Member of the RAS A. G. Tonevitsky
c
,
and Academician E. D. Sverdlov
a
Received December 24, 2010
DOI: 10.1134/S1607672911020153
a
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry,
Russian Academy of Sciences,
ul. MiklukhoMaklaya 16/10, Moscow, 117997 Russia
b
Blokhin Cancer Research Center,
Russian Academy of Medical Sciences,
Kashirskoe sh. 24, Moscow, 115478 Russia
c
AllRussia Institute of Physical Culture and Athletics,
Elizavetinskii per. 10, Moscow, 105005 Russia
BIOCHEMISTRY, BIOPHYSICS
AND MOLECULAR BIOLOGY