Abstract
BackgroundCancer remains one of the leading causes of death worldwide, despite the possibilities to detect early onset of the most common cancer types. The search for the optimal therapy is complicated by the cancer diversity within tumors and the unsynchronized development of cancerous cells. Therefore, it is necessary to characterize cancer cell populations after treatment has been applied, because cancer recurrence is not rare. In our research, we concentrated on small cancer cell subpopulation (microcells) that has a potential to be cancer resistance source. Previously made experiments has shown that these cells in small numbers form in specific circumstances after anticancer treatment.MethodsIn experiments described in this research, the anticancer agents’ paclitaxel and doxorubicin were used to stimulate the induction of microcells in fibroblast, cervix adenocarcinoma, and melanoma cell lines. Mainly for the formation of microcells in melanoma cells. The drug-stimulated cells were then characterized in terms of their formation efficiency, morphology, and metabolic activity.ResultsWe observed the development of cancer microcells and green fluorescent protein (GFP) transfection efficiency after stress. In the time-lapse experiment, we observed microcell formation through a renewal process and GFP expression in the microcells. Additionally, the microcells were viable after anticancer treatment, as indicated by the nicotinamide adenine dinucleotide hydrogen phosphate (NADPH) enzyme activity assay results. Taken together, these findings indicate that cancer microcells are viable and capable of resisting the stress induced by anticancer drugs, and these cells are prone to chemical substance uptake from the environment.ConclusionMicrocells are not only common to a specific cancer type, but can be found in any tumor type. This study could help to understand cancer emergence and recurrence. The appearance of microcells in the studied cancer cell population could be an indicator of the individual anticancer therapy effectiveness and patient survival.
Highlights
Cancer remains one of the leading causes of death worldwide, despite the possibilities to detect early onset of the most common cancer types
Using 8-anilinonaphthalene-1-sulfonic acid (ANS)- Ethidium bromide (EtBr), nicotinamide adenine dinucleotide hydrogen phosphate NADPH assay, and time-lapse experiments with green fluorescent protein (GFP) transfection, we have shown that the microcells are viable and metabolically active
Cell lines Human cervical carcinoma (HeLa), SK-MEL-28 melanoma, and HS-68 cell lines were acquired from the American Type Culture Collection (ATCC) and maintained in a culture medium consisting of Dulbecco’s modified Eagle’s medium (DMEM; Thermo Scientific, IL, USA) supplemented with 10% fetal bovine serum (FBS)
Summary
Cancer remains one of the leading causes of death worldwide, despite the possibilities to detect early onset of the most common cancer types. Buiķis and colleagues have previously shown that human sarcoma cell cultures treated with thiophosphamide (ThioTEPA), a chemotherapeutic agent used to treat breast, ovarian and bladder cancers [3, 4] exhibit the development of unusual small-sized cells, called microcells. These cells are round or oval, with a small amount of cytoplasm, and are intensively stained by bromo-2deoxyuridine (BrdU) that is thymidine analog and it incorporates into DNA and shows cells in S-phase and methyl green-pyronine that stains nucleic acids both in DNA and RNA [4,5,6]. One defective macrocell may produce one or several microcells [4, 8]
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