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May 22-23, 2019 | Rome, Italy
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IMMUNOLOGY
AND CANCER THERAPY
2
nd
Global Summit on
Immunotherapy 2019
Immunology Case Reports | Volume 3
EXPERIENCE OF THE USE OF NSC-631570 (UKRAIN) IN THE TREATMENT OF MALIG-
NANT MELANOMA
Wassil Nowicky
1
and
Larysa Skivka
2
1
Ukrainian Anti-Cancer Institute, Austria
2
Taras Shevchenko National University of Kyiv, Austria
Introduction:
Malignant melanoma is one of the most deadly skin cancers. At the early stages of melanoma
development the patients are treated surgically, but the advanced disease is virtually incurable. Numerous
clinical investigations have been conducted to improve efficiency of melanoma treatment. Nevertheless, biol-
ogists and clinicians continue working on new possible methodology of treatment and keep up to search new
therapeutic agents as drug resistance is a commonly observed problem. NSC-631570 is an anti-cancer agent
created on the basis of alkaloids from the plant
Chelidonium majus
. For more than 20 years NSC-631570 had
been used for cancer treatment. Monotherapy and combined application of NSC-631570 are successfully used
for the treatment of malignant melanoma since 1996.
Aim:
The purpose of this study is to describe the experience of the use of NSC-631570 in the treatment of ma-
lignant melanoma and to disclose of some mechanisms of the preparation action.
Materials & Methods:
B-16 melanoma cells of C57BL/6 mice were kindly supplied by the Bank of Cell Cultures
and Transplantable Experimental Tumors of R.E. Kavetsky Institute of Experimental Pathology, Oncology and
Radiobiology (Kyiv, Ukraine) MM-4 cells, exhibiting a relatively low metastatic potential, were established from
the primary lesion of subcutaneously injected B16 cells. MM-4M2 cell lines established by two sequential pas-
sages of lung metastases of MM-4 cells after intravenous injection are highly metastatic. Cells were cultured
in
vitro
in Dulbecco’s Modified Eagle Medium (DMEM; Sigma, St. Louis, MO, USA) supplemented with 10% fetal
calf serum (FCS), penicillin (100 U/ml) and streptomycin (100 μg/ml) at 37ºC in 5% CO2. To determine the effects
of NSC-631570 on cell viability, cells were treated with NSC6-31570 at the different concentrations (1.6, 3.2, 6.4,
12.5, 25, 50, 100 and 200 μg/ml) for 24hrs and 48hrs periods. Cell viability was determined by MTT test. HMGB1
level in conditioned medium was evaluated by ELISA. TAP mRNA was examined by RT-PCR and TAP protein in
ELL Lysates was determined by Western blot.
Results:
Treatment of B16 melanoma cells with NSC-631570 at apoptogenic concentrations induced dose-de-
pendent tumor cell death accompanied by dose dependent release of HMGB1, more in high-metastasizing
cells. The levels of HMGB1 in the cell probes treated with the drug exhibited strong correlation with the levels
of cell death. Author found a significant increase in the number of mRNA for TAP1 in melanoma B16 cells after
treatment with NSC-631570 at the non-apoptogenic concentration, whereas only a trace quantity of the TAP1
mRNA was found in untreated cells: Treatment of melanoma B16 cells with NSC-631570 increased expression
of mRNA encoding TAP2 by ~3-fold (p< 0.05); increase of TAP1-proteins expression was confirmed by Western
blot analysis. It is known that correction of TAP1 and/or TAP2 defects in B16 mouse melanoma enhances the
cell surface expression of MHC class I molecules and significantly reduces the rate of subcutaneous tumor
growth and pulmonary metastatic burden.
Wassil Nowicky et al., Immunol Case Rep 2019, Volume 3