Poster presentation
325
EFFECT OF Cuscuta europeae GLYCOPROTEINS
ON CANCER CELLS
Z.S. Khashimova, K.A. Kakhorova, M.K. Salakhutdinova,
М.S. Аbdullahodzaeva, Yu.I. Oshchepkova
Institute of Bioorganic Chemistry, Academy of Sciences, Tashkent, Uzbekistan
Despite the existence of a number of theories of ontogenesis, including those aimed
at creating new anticancer drugs, there are many outstanding oncological problems.
This indicates not only the complexity, but also the
need to develop new approaches,
both to cancer chemotherapy and to understanding the overall problem. One such
approach is early cancer diagnosis and treatment. So far, among oncological diseases,
cervical cancer remains one of the most pressing problems in oncogynecology.
Professor M.S. Abdullahodzhaeva and co-workers
found distinctive dispersed
luminous particles (DLP) in polarized light on the surface of epithelial cells of the
cervix of cancer patients. Such dispersed luminous particles (DLP) appeared only in
cases of diagnosed cervical cancer. Normally, as well as in inflammatory or dystrophic
process in the cervix, these particles were not detected.
In this regard, the purpose of
this work was to investigate DLP in cultured cancer cells and study the effect of various
anticancer drugs on them. A transplanted HeLa cell culture was used to study the nature
of DLP.
HeLa cells in an amount of 1 million/ml were placed to 24-well plates and cultured in
5 ml of DMEM growth medium containing an antimycotic antibiotic, L-glutamine, 10%
fetal calf serum in a CO2 incubator for 24 hours. Then the cells were removed from the
surface plastic with versin,
washed with culture medium, and the cell suspension was
placed to a defatted glass slide, on which a non-fixed smear was made with another glass
slide. The smear dried in air for 3 to 5 minutes. After that,
the glass slide was placed
smear down on the stage of the «Axiovert 40 MAT» reflective optical microscope, where
the DLP were visualized. When viewed under a microscope in polarized light, 40% of the
cells showed DLP, i.e. not all cells in transplanted HeLa cancer culture contain such
structures. Subsequently, we studied clinical antitumor drugs with a known mechanism
of action, namely, methotrexate and cisplatin on HeLa cell culture. To do this, HeLa
cells were seeded into 96-well plates in an amount of 20000–30000 cells/mL in 100 μL
of DMEM medium with 10% fetal calf serum and cultured at 37°C in a CO2 incubator.
A day later, the preparations were administered at doses of 100, 10, and 1 μg/ml per 100
μl of the medium, and the cells were cultured for 24 hours. Cells without drug exposure
served as control. Cytotoxic activity was assessed by incorporation of MTT into cells. A
high cytotoxic activity of drugs (70-90%) on HeLa cells was established. Microscopic
examination revealed DLP in control cells, but no intracellular DLP were found in the
experimental samples.
Since DLP is observed on the surface of epithelial cancer cells, we studied the effect
of carbohydrate-containing proteins on HeLa cells. Proteins
were obtained from the
seeds of Cuscuta europeae by extraction with 80% saline ammonium sulfate. The
content of carbohydrates was determined and the hemagglutinating activity of the
protein was established. Proteins exhibit high cytotoxic activity on HeLa cells.
Microscopic examination did not reveal DLP in the experimental samples, in contrast to
the control ones.