ISSN 0564-3783  

Main page
Information to authors
Editorial board
Mobile version

In Ukrainian

Export citations

Research of stem tumor cells subpopulation on models of breast cancer

Herheliuk T., Perepelytsina O., Chmelnytskaia Y., Kuznetsova G., Dzjubenko N., Raksha N., Gorbach O., Sydorenko M.


SUMMARY. Heterogenity of the tumor population, the presence of tumor stem cells in it, is one of the reasons for the resistance of tumors to antitumor therapy, recurrence and metastasis, as well as the complexity of treatment of the cancer. The aim of the study was to enrich of the breast adenocarcinoma cells, MCF-7, multicellular tumor spheroids (MTS) with cancer stem cells, enriched MTS (eMTS). Other aim was to investigate the resulting subpopulation of CSC in MTS by biochemical, immunological and cytological methods. According to the results of the study, it was found that in the conditions of lack of nutrient medium, with the addition of certain growth factors, the percentage of CSC in the cell population of MTS increased significantly. The obtained results demonstrated increasing of the CSC subpopulation. It was indicated according to biochemical, cytological and immunological methods simultaneously. Thus, the percentage of CD133+ cells increased from 12.47 to 82.08 %, Nestin+ from 31.3 to 82.58 %. According to immunohistochemical staining data the expression of
other markers of CSC: CD44, CD133, bmi1, also increased. The activity of aldehyde dehydrogenase in MCF-7 cells in monolayer culture was 0.07 mol/mg protein per minute and increased to 1.58 mol/mg of protein per minute in eMTS. The activity of glucose-6-phosphate dehydrogenase (G6FDG) in MCF-7 under conditions of monolayer growth was 934.6 148.3 × 106 mol/mg of protein per minute. At the same time in the enriched by CSC MTS the activity of G6FDG increased more than in 1.5 times. The activity of the lactate dehydrogenase (LDH) in MCF-7 cells in monolayer culture was 65.12 1.28 mol/mg of protein per minute, and in eMTS, decreased in 5.5 times. Thus, based on the analysis of the obtained data, we can assume that under conditions of enrichment of the tumor population by CSC the receptor and energy profile of MCF-7 cells changed. So, MTS are approaching to characteristics of metastatic micronode, and tumor cells are approaching to cancer stem cells.

Tsitologiya i Genetika 2022, vol. 56, no. 4, pp. 24-38

  1. Department of Biotechnical Problems of Diagnostics, Institute for Problems of Cryobiology and Cryomedicine, National Academy of Science of Ukraine, 03028, Kyiv, Ukraine
  2. Educational and Scientific Centre Institute of Biology and Medicine Kyiv, Ukraine
  3. National Cancer Institute of Ukraine, 03022, Kyiv, Ukraine

E-mail: olenaquil, vbpd-ipkk

Herheliuk T., Perepelytsina O., Chmelnytskaia Y., Kuznetsova G., Dzjubenko N., Raksha N., Gorbach O., Sydorenko M. Research of stem tumor cells subpopulation on models of breast cancer, Tsitol Genet., 2022, vol. 56, no. 4, pp. 24-38.

In "Cytology and Genetics":
T. S. Herheliuk, O. M. Perepelytsina, Yu. M. Chmelnytska, G. M. Kuznetsova, N. V. Dzjubenko, N. G. Raksha, O. I. Gorbach & M. V. Sydorenko Study of Cancer Stem Cell Subpopulations in Breast Cancer Models, Cytol Genet., 2022, vol. 56, no. 4, pp. 331342
DOI: 10.3103/S0095452722040041


Baccelli, I. and Trumpp, A., The evolving concept of cancer and metastasis stem cells, J. Cell Biol., 2012, vol. 198, pp. 281293.

Bapat, S., Mali, A., Koppikar, C., et al., Stem and progenitor-like cells contribute to the aggressive behavior of human epithelial ovarian cancer, Cancer Res., 2005, vol. 65, pp. 30253029.

Bjerkvig, R., Spheroid Culture in Cancer Research, Boca Raton: CRC Press, 1992.

Borlle, L., Dergham, A., Wund, Z., et al., Salinomycin decreases feline sarcoma and carcinoma cell viability when combined with doxorubicin, BMC Vet. Res., 2019, vol. 15, no. 1.

Brugnoli, F., Grassilli, S., Al-Qassab, Y., et al., CD133 in breast cancer cells: more than a stem cell marker, J. Oncol., 2019, vol. 2019, art. ID 7512632.

Colak, S. and Medema, J., Cancer stem cells important players in tumor therapy resistance, FEBS J., 2014, vol. 281, no. 21, pp. 47794791.

Collins, A., Berry, P., Hyde, C., et al., Prospective identification of tumorigenic prostate cancer stem cells, Cancer Res., 2005, vol. 65, pp. 1094610951.

Cui, J., Shi, M., Xie, D., et al., FOXM1 promotes the Warburg effect and pancreatic cancer progression via transactivation of LDHA expression, Clin. Cancer Res., 2014, vol. 20, no. 10, pp. 25952606.

Ehrmann, J., Kolar, Z., and Mokry, J., Nestin as a diagnostic and prognostic marker: immunohistochemical analysis of its expression in different tumours, J. Clin. Pathol., 2005, vol. 58, no. 2, pp. 222223.

Feng, Y., Xiong, Y., and Qiao, T., Lactate dehydrogenase A: A key player in carcinogenesis and potential target in cancer therapy, Cancer Med., 2018, vol. 7, no. 12.

Ghanbari Movahed, Z., Rastegari-Pouyani, M., Mohammadi, M., et al., Cancer cells change their glucose metabolism to overcome increased ROS: One step from cancer cell to cancer stem cell?, Biomed. Pharmacother., 2019, vol. 112, art. ID 108690.

Giatromanolaki, A., Sivridis, E., Gatter, K., et al., Lactate dehydrogenase 5 (LDH-5) expression in endometrial cancer relates to the activated VEGF/VEGFR2(KDR) pathway and prognosis, Gynecol. Oncol., 2006, vol. 103, no. 3, pp. 912918.

He, Q., Luo, X., Wang, K., et al., Isolation and characterization of cancer stem cells from high-grade serous ovarian carcinomas, Cell. Physiol. Biochem., 2014, vol. 33, no. 1, pp. 173184.

Herheliuk, T., Perepelytsina, O., Ugnivenko, A., et al., Investigation of multicellular tumor spheroids enriched for a cancer stem cell phenotype, Stem Cell Invest., 2019, vol. 6, art. ID 21.

Hermann, P., Huber, S., Herrler, T., et al., Distinct populations of cancer stem cells determine tumor growth and metastatic activity in human pancreatic cancer, Cell Stem Cell, 2007, vol. 1, no. 3, pp. 313323.

Hockmair, M., Rath, B., Klameth, L., et al., Effects of salinomycin and niclosamide on small cell lung cancer and small cell lung cancer circulating tumor cell lines, Invest. New Drugs, 2020, vol. 38, no. 4, pp. 46955.

Jiang, P., Du, W., and Wu, M., Regulation of the pentose phosphate pathway in cancer, Protein Cell, 2014, vol. 5, pp. 592602.

Jiang, W., Zhou, F., Li, N., et al., FOXM1-LDHA signaling promoted gastric cancer glycolytic phenotype and progression, Int. J. Clin. Exp. Pathol., 2015, vol. 8, no. 6, pp. 67566763.

Karakaya, H. and Ozkul, K., Measurement of glucose-6-phosphate dehydrogenase activity in bacterial cell-free extracts, Bio-Protoc., 2016, vol. 6, no. 19, art. ID e1949.

Ketola, K., Hilvo, M., Hyötyläinen, T., Vuoristo, A., et al., Salinomycin inhibits prostate cancer growth and migration via induction of oxidative stress, Brit. J. Cancer, 2012, vol. 106, pp. 99106.

Kim, Y., Siegler, E., Siriwon, N., and Wang, P., Therapeutic strategies for targeting cancer stem cells, J. Cancer Metastasis Treat., 2016, vol. 2, pp. 233242.

Kleeberger, W., Bova, G.S., and Nielsen, M.E., Roles for the stem cell associated intermediate filament Nestinin prostate cancer migration and metastasis, Cancer Res., 2007, vol. 67, no. 19, pp. 91999206.

Koukourakis, M., Kakouratos, C., and Kalamida, D., Hypoxia-inducible proteins HIF1α and lactate dehydrogenase LDH5, key markers of anaerobic metabolism, relate with stem cell markers and poor post-radiotherapy outcome in bladder cancer, Int. J. Radiat. Biol., 2016, vol. 92, no. 7, pp. 353363.

Krupkova, Jr., Loja, T., Zambo, I., and Veselska, R., Nestin expression in human tumors and tumor cell lines, Neoplasma, 2010, vol. 4, pp. 291298.

Kumar, V. and Gill, K.D., Determination of total lactate dehydrogenase activity in serum sample, in Basic Concepts in Clinical Biochemistry, A Practical Guide, Springer-Verlag, 2018, pp. 129130.


Kurpinska, A., Suraj, J., Bonar, E., et al., Proteomic characterization of early lung response to breast cancer metastasis in mice, Exp. Mol. Pathol., 2019, vol. 407, pp. 129140.

Ma, L., Lai, D., Liu, T., et al., Cancer stem-like cells can be isolated with drug selection in human ovarian cancer cell line SKOV3, Acta Biochim. Biophys. Sin., 2010, vol. 42, no. 9, pp. 593602.

Mukherjee, D. and Ahmad, R., Glucose-6-phosphate dehydrogenase activity during N'-nitrosodiethylamine-induced hepatic damage, Ach. Life Sci., 2015, vol. 9, pp. 5156.

Naujokat, C., Salinomycin in cancer: A new mission for an old agent, Mol. Med. Rep., 2015, vol. 3, no. 4, pp. 555559.

Neradil, J. and Veselska, R., Nestin as a marker of cancer stem cells, Cancer Sci., 2015, vol. 106, no. 7, pp. 803811.

Patra, K. and Hay, N., The phosphate pathway and cancer, Trends Biochem., 2014, vol. 39, pp. 347354.

Piras, F., Perra, M.T., Murtas, D., et al., The stem cell marker nestin predicts poor prognosis in human melanoma, Oncol. Rep., 2010, vol. 23, no. 1, pp. 1724.

Ramos-Martinez, J., The regulation of the pentose phosphate pathway: Remember Krebs, Arch. Biochem. Biophys., 2017, vol. 614, pp. 5052.

Rappa, G., Fodstad, O., and Lorico, A., The stem cell-associated antigen CD133 (Prominin-1) is a molecular therapeutic target for metastatic melanoma, Stem Cells, 2008, vol. 26, no. 12, pp. 30083017.

Resham, K., Patel, P., Thummuri, D., et al., Preclinical drug metabolism and pharmacokinetics of salinomycin, a potential candidate for targeting human cancer stem cells, Chem.-Biol. Interact., 2015, vol. 240, pp. 146152.

Sant, S., Johnston, P., et al., The production of 3D tumor spheroids for cancer drug discovery, Drug Discovery Today: Technol., 2017, vol. 23, pp. 2736.

Schneider, M., Huber, J., Hadaschik, B., et al., Characterization of colon cancer cells: a functional approach characterizing CD133 as a potential stem cell marker, BMC Cancer, 2012, vol. 12, art. ID 96.

Singh, Sh., Clarke, I., Terasaki, M., et al., Identification of a cancer stem cell in human brain, Cancer Res., 2003, vol. 63, no. 18, pp. 58215828.

Strojnik, T., Rosland, G.V., Sakariassen, P.O., et al., Neural stem cell markers, nestin and musashi proteins, in the progression of human glioma: correlation of nestin with prognosis of patient survival, Surg. Neurol., 2007, vol. 68, no. 2, pp. 133143.

Su, Y., Yu, Q., Wang, X., et al., JMJD2A promotes the Warburg effect and nasopharyngeal carcinoma progression by transactivating LDHA expression, BMC Cancer, 2007, vol. 17, art. ID 477.

Talaiezadeh, A., Shahriari, A., Tabandeh, M., et al., Kinetic characterization of lactate dehydrogenase in normal and malignant human breast tissues, Cancer Cell Int., 2015, vol. 15, art. ID 19.

Tang, Q., Zhao, Z.-Q., Li, J.-C., Liang, Y., et al., Salinomycin inhibits osteosarcoma by targeting its tumor stem cells, Cancer Lett., 2011, vol. 311, pp. 113121.

Taniguchi, M., Mori, N., and Iramina, C., Elevation of glucose 6-phosphate dehydrogenase activity induced by amplified insulin response in low glutathione levels in rat liver, Sci. World J., 2016, vol. 2016, art. ID 6382467.

Teranishi, N., Naito, Z., Ishiwata, T., et al., Identification of neovasculature using nestin in colorectal cancer, Int. J. Oncol., 2007, vol. 30, no. 3, pp. 593603.

Tropepe, V., Alton, K., Sachewsky, N., et al., Neurogenic potential of isolated precursor cells from early post-gastrula somitic tissue, Stem Cells Dev., 2009, vol. 18, no. 10, pp. 15331542.

Vassalli, G., Aldehyde Dehydrogenases: not just markers, but functional regulators of stem cells, Stem Cells Int., 2019, vol. 2019, art. ID 3904645.

Versini, A., Colombeau, L., and Hienzsch, A., Salinomycin derivatives kill breast cancer stem cells by lysosomal iron targeting, Chem. - Eur. J., 2020, vol. 26, no. 33.

Wang, H., Zhang, H., Zhu, Y., et al., Anticancer mechanisms of salinomycin in breast cancer and its clinical applications, Front. Oncol., 2021.

Wang, Y., Effects of salinomycin on cancer stem cell in human lung adenocarcinoma A549 cells, Med. Chem., 2011, vol. 7, no. 2, pp. 106111.

Wong, T., Che, N., and Ma, S., Reprogramming of central carbon metabolism in cancer stem cells, Biochim. Biophys. Acta, Mol. Basis Dis., 2017, vol. 1863, pp. 17281738.

Yin, A.H., Miraglia, S., Zanjani, E.D., et al., AC133, a novel marker for human hematopoietic stem and progenitor cells, Blood, 1997, vol. 90, no. 12, pp. 50025012.

Zdralevic, M., Marchiq, I., Cunhade, P., et al., Metabolic plasiticy in cancersdistinct role of glycolytic enzymes GPI, LDHs or membrane transporters MCTs, Front. Oncol., 2017.

Zhang, C., Tian, Y., Song, F., et al., Salinomycin inhibits the growth of colorectal carcinoma by targeting tumor stem cells, Oncol. Rep., 2015.

Zhao, Z., Lu, P., and Zhang, H., Nestin positively regulates the Wnt/β-catenin pathway and the proliferation, survival and invasiveness of breast cancer stem cells, Breast Cancer Res., 2014, vol. 16, art. ID 408.

Zhi, Q., Chen, X., Ji, J., Zhang, J., et al., Salinomycin can effectively kill ALDHhigh stem-like cells on gastric cancer, Biomedicine & Pharmacotherapy, 2011, vol. 65, no. 7, pp. 509515.

Copyright© ICBGE 2002-2023 Coded & Designed by Volodymyr Duplij Modified 28.11.23