In vitro Cytotoxic Effect of Ethanol and Dimethyl Sulfoxide on Various Human Cell Lines

Elif Ayazoglu Demir; Selim Demir; Yüksel Aliyazıcıoğlu

doi:10.18016/ksutarimdoga.vi.702702

Research Article

Etanol ve Dimetil Sülfoksidin Çeşitli İnsan Hücre Hatları Üzerindeki in vitro Sitotoksik Etkisi

Year 2020, Volume: 23 Issue: 5, 1119 - 1124, 31.10.2020

Elif Ayazoglu Demir Selim Demir , Yüksel Aliyazıcıoğlu

https://doi.org/10.18016/ksutarimdoga.vi.702702

Cited By: 7

Abstract

Literatürde etanol ve dimetil sülfoksit (DMSO)’in bazı hücre hatları üzerindeki sitotoksik etkileri gösterilmiş olmasına rağmen, bu çözücülerin melanoma (VMM917), akciğer kanseri (A549), kolon kanseri (WiDr), normal kolon (CCD 841 CoN) ve fibroblast (BJ) hücreleri üzerindeki sitotoksik etkilerini belirleyen bir çalışma bulunmamaktadır. Bu çalışmanın amacı, etanol ve DMSO'nun yukarıda belirtilen hücrelere ek olarak insan meme (MCF-7), karaciğer (HepG2) ve serviks (HeLa) kanser hücreleri üzerindeki sitotoksik etkilerini belirlemektir. Bu amaçla, hücreler farklı konsantrasyonlarda (%0.1, %0.2, %0.4, %0.6, %0.8, %1, %2 ve % 4, hacim/hacim) etanol ve DMSO ile muamele edilmiş ve daha sonra MTT prosedürü uygulanmıştır. Elde edilen sonuçlar, etanol ve DMSO’nun incelenen tüm hücrelerde doza bağlı sitotoksisite sergilediğini göstermiştir. DMSO ve etanole en duyarlı hücreler sırasıyla WiDr ve VMM917 iken, en dirençli hücreler sırasıyla BJ ve A549 olarak belirlenmiştir. Sonuçlar, her bir hücre hattında etanol ve DMSO'nun sitotoksik etki sergilediği konsantrasyon aralığının farklı olduğunu ortaya koymuştur. Yanlış pozitif ve negatif sonuçlara neden olmamak için, hücre kültürü çalışmalarında öncelikle kullanılacak çözücülerin sitotoksik etkilerinin oluşmadığı konsantrasyon aralığı belirlenmelidir.

Keywords

Hücre kültürü, Sitotoksisite, Etanol, Dimetil sülfoksit, MTT yöntemi

References

Aliyazicioglu R, Demir S, Badem M, Sener SO, Korkmaz N, Demir EA, Ozgen U, Karaoglu SA, Aliyazicioglu Y, 2017. Antioxidant, antigenotoxic, antimicrobial activities and phytochemical analysis of Dianthus carmelitarum. Rec Nat Prod, 11(3): 270-284.
Aliyazicioglu Y, Demir S, Yaman SO, Sener SO, Demir EA, Aliyazicioglu R, Turan I, 2019. Phytochemical analysis of Dorycnium pentaphyllum and its antiproliferative effect on cervix cancer cells. KSU J Agric Nat, 22(Suppl 2): 365-373.
Ben Trivedi A, Kitabatake N, Doi E, 1990. Toxicity of dimethyl sulfoxide as a solvent in bioassay system with HeLa cells evaluated colorimetrically with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide. Agric Biol Chem, 54(11): 2961-2966.
Berg KCG, Eide PW, Eilertsen IA, Johannessen B, Bruun J, Danielsen SA, Bjørnslett M, Meza-Zepeda LA, Eknæs M, Lind GE, Myklebost O, Skotheim RI, Sveen A, Lothe RA, 2017. Multi-omics of 34 colorectal cancer cell lines-a resource for biomedical studies. Mol Cancer, 16(1): 116.
Casañas-Sánchez V, Pérez JA, Quinto-Alemany D, Díaz M, 2016. Sub-toxic ethanol exposure modulates gene expression and enzyme activity of antioxidant systems to provide neuroprotection in hippocampal HT22 cells. Front Physiol, 7: 312.
Clave S, Joya X, Salat-Batlle J, Garcia-Algar O, Vall O, 2014. Ethanol cytotoxic effect on trophoblast cells. Toxicol Lett, 225(2): 216-221.
Da Violante G, Zerrouk N, Richard I, Provot G, Chaumeil JC, Arnaud P, 2002. Evaluation of the cytotoxicity effect of dimethyl sulfoxide (DMSO) on Caco2/TC7 colon tumor cell cultures. Biol Pharm Bull, 25(12): 1600-1603.
de Abreu Costa L, Henrique Fernandes Ottoni M, Dos Santos MG, Meireles AB, Gomes de Almeida V, de Fátima Pereira W, Alves de Avelar-Freitas B, Eustáquio Alvim Brito- Melo G, 2017. Dimethyl sulfoxide (DMSO) decreases cell proliferation and TNF-α, IFN-γ, and IL-2 cytokines production in cultures of peripheral blood lymphocytes. Molecules, 22(11): E1789.
Demir S, Turan I, Aliyazicioglu Y, 2018a. Cytotoxic effect of Rhododendron luteum leaf extract on human cancer cell lines. KSU J Agric Nat, 21(6): 950-956.
Demir S, Turan I, Aliyazicioglu R, Ozer Yaman S, Aliyazicioglu Y, 2018b. Primula vulgaris extract induces cell cycle arrest and apoptosis in human cervix cancer cells. J Pharm Anal, 8(5): 307-311.
Demir S, Turan I, Aliyazicioglu Y, 2019a. Antioxidant properties of Primula vulgaris flower extract and its cytotoxic effect on human cancer cell lines. KSU J Agric Nat, 22(1): 78-84.
Demir S, Turan I, Misir S, Aliyazicioglu Y, 2019b. Selective cytotoxic effect of Dorycnium pentaphyllum extract on human breast, liver, and lung cancer cells. KSU J Agric Nat, 22(3): 473-479.
Demir S, Yaman SO, Sener SO, Demir EA, Aliyazicioglu R, Ozgen U, Mentese A, Deger O, Aliyazicioglu Y, 2020. Dorycnium pentaphyllum extract has antiproliferative effect on human cervix and colon cancer cells. Nutr Cancer, 72(3): 504-512.
Ferreira D, Silva AP, Nobrega FL, Martins IM, Barbosa-Matos C, Granja S, Martins SF, Baltazar F, Rodrigues LR, 2019. Rational identification of a colorectal cancer targeting peptide through phage display. Sci Rep, 9: 3958.
Forman S, Kas J, Fini F, Steinberg M, Ruml T, 1999. The effect of different solvents on the ATP/ADP content and growth properties of HeLa cells. J Biochem Mol Toxicol, 13(1): 11-15.
Galvao J, Davis B, Tilley M, Normando E, Duchen MR, Cordeiro MF, 2014. Unexpected low-dose toxicity of the universal solvent DMSO. FASEB J, 28(3): 1317-1330.
Gazdar AF, Girard L, Lockwood WW, Lam WL, Minna JD, 2010. Lung cancer cell lines as tools for biomedical discovery and research. J Natl Cancer Inst, 102(17): 1310-1321.
Hajighasemi F, Tajik S, 2017. Assessment of cytotoxicity of dimethyl sulfoxide in human hematopoietic tumor cell lines. IJBC, 9(2): 48-53.
Hebling J, Bianchi L, Basso FG, Scheffel DL, Soares DG, Carrilho MRO, Pashley DH, Tjäderhane L, de Souza Costa CA, 2015. Cytotoxicity of dimethyl sulfoxide (DMSO) in direct contact with odontoblast-like cells. Dent Mater, 31(4): 399-405.
Jamalzadeh L, Ghafoori H, Sariri R, Rabuti H, Nasirzade J, Hasani H, Aghamaali MR, 2016. Cytotoxic effects of some common organic solvents on MCF-7, RAW-264.7 and human umbilical vein endothelial cells. Avicenna J Med Biochem, 4(1): e33453.
Kade S, Herzog N, Schmidtke KU, Küpper JH, 2016. Chronic ethanol treatment depletes glutathione regeneration capacity in hepatoma cell line HepG2. J Cell Biotech, 1(2): 183-190.
Liu S, Gao G, Yan D, Chen X, Yao X, Guo S, Li G, Zhao Y, 2017. Effects of miR-145-5p through NRAS on the cell proliferation, apoptosis, migration, and invasion in melanoma by inhibiting MAPK and PI3K/AKT pathways. Cancer Med, 6(4): 819-833.
Mikami K, Haseba T, Ohno Y, 1997. Ethanol induces transient arrest of cell division (G2+M block) followed by G0/G1 block: dose effects of shortand longer-term ethanol exposure on cell cycle and cell functions. Alcohol Alcohol, 32(2): 145-152.
Misir S, Aliyazicioglu Y, Demir S, Turan I, Yaman SO, Deger O, 2018. Antioxidant properties and protective effect of Turkish propolis on t-BHP-induced oxidative stress in foreskin fibroblast cells. IJPER, 52(1): 94-100.
Misir S, Aliyazicioglu Y, Demir S, Turan I, Hepokur C, 2020. Effect of Turkish propolis on miRNA expression, cell cycle, and apoptosis in human breast cancer (MCF-7) cells. Nutr Cancer, 72(1):133-145.
Mosmann T, 1983. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods, 65: 55-63.
Schäuble S, Klement K, Marthandan S, Münch S, Heiland I, Schuster S, Hemmerich P, Diekmann S, 2012. Quantitative model of cell cycle arrest and cellular senescence in primary human fibroblasts. PLoS One, 7(8): e42150.
Singh M, McKenzie K, Ma X, 2017. Effect of dimethyl sulfoxide on in vitro proliferation of skin fibroblast cells. J Biotech Res, 8: 78-82.
Timm M, Saaby L, Moesby L, Hansen EW, 2013. Considerations regarding use of solvents in in vitro cell based assays. Cytotechnology, 65: 887-894.
Turan I, Demir S, Kilinc K, Aliyazicioglu Y, Alver A, Misir S, Ozer Yaman S, Akbulut K, Mentese A, Deger O, 2017. Morus rubra extract induces G1 cell cycle arrest and apoptosis in human lung and prostate cancer cells. IJPER, 51(1): 51-58.
Turan I, Demir S, Kilinc K, Yaman SO, Misir S, Kara H, Genc B, Mentese A, Aliyazicioglu Y, Deger O, 2018. Cytotoxic effect of Rosa canina extract on human colon cancer cells through repression of telomerase expression. J Pharm Anal, 8(6): 394-399.
Wu D, Bahaerguli K, Wu GR, Sheng L, 2010. Studies on the effect of DMSO as a solvent mediator on cell growth and viability. J Xinjiang Med Univ, 33: 489-491.
Yeo EJ, Lim HK, Park SC, 2000. Effect of short-term ethanol on the proliferative response of Swiss 3T3 cells to mitogenic growth factors. Exp Mol Med, 32(3): 161-169.
Yi X, Liu M, Luo Q, Zhuo H, Cao H, Wang J, Han Y, 2017. Toxic effects of dimethyl sulfoxide on red blood cells, platelets, and vascular endothelial cells in vitro. FEBS Open Bio, 7: 485-494.
Yuan C, Gao J, Guo J, Bai L, Marshall C, Cai Z, Wang L, Xiao M, 2014. Dimethyl sulfoxide damages mitochondrial integrity and membrane potential in cultured astrocytes. PLoS One, 9(9): e107447

In vitro Cytotoxic Effect of Ethanol and Dimethyl Sulfoxide on Various Human Cell Lines

Year 2020, Volume: 23 Issue: 5, 1119 - 1124, 31.10.2020

Elif Ayazoglu Demir Selim Demir , Yüksel Aliyazıcıoğlu

https://doi.org/10.18016/ksutarimdoga.vi.702702

Cited By: 7

Abstract

Although the cytotoxic effects of ethanol and dimethyl sulfoxide (DMSO) on some cell lines have been shown in the literature, there is no study about the cytotoxic effects of these solvents on common used human cell lines, such as melanoma (VMM917), lung cancer (A549), colon cancer (WiDr), normal colon (CCD 841 CoN) and fibroblast (BJ) cells. The aim of this study was to determine the cytotoxic effects of ethanol and DMSO on human breast (MCF-7), liver (HepG2) and cervix (HeLa) cancer cells in addition to above mentioned cells. For this purpose, the cells were treated with different concentrations (0.1%, 0.2%, 0.4%, 0.6%, 0.8%, 1%, 2%, and 4%, v/v) of ethanol and DMSO and then subjected to MTT assay. According to the results, ethanol and DMSO exhibited dose-dependent cytotoxicity in all cells studied. The most DMSO and ethanol sensitive cells were WiDr and VMM917, while the most resistant cells were determined as BJ and A549, respectively. The results revealed that the concentration range in which ethanol and DMSO exhibited cytotoxic effect in each cell line is different. In order not to cause false positive and negative results, the concentration range in which the solvents used in cell culture studies do not have cytotoxic effects should be determined.

Keywords

Cell culture, Cytotoxicity, Ethanol, Dimethyl sulfoxide, MTT assay

References

Aliyazicioglu R, Demir S, Badem M, Sener SO, Korkmaz N, Demir EA, Ozgen U, Karaoglu SA, Aliyazicioglu Y, 2017. Antioxidant, antigenotoxic, antimicrobial activities and phytochemical analysis of Dianthus carmelitarum. Rec Nat Prod, 11(3): 270-284.
Aliyazicioglu Y, Demir S, Yaman SO, Sener SO, Demir EA, Aliyazicioglu R, Turan I, 2019. Phytochemical analysis of Dorycnium pentaphyllum and its antiproliferative effect on cervix cancer cells. KSU J Agric Nat, 22(Suppl 2): 365-373.
Ben Trivedi A, Kitabatake N, Doi E, 1990. Toxicity of dimethyl sulfoxide as a solvent in bioassay system with HeLa cells evaluated colorimetrically with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide. Agric Biol Chem, 54(11): 2961-2966.
Berg KCG, Eide PW, Eilertsen IA, Johannessen B, Bruun J, Danielsen SA, Bjørnslett M, Meza-Zepeda LA, Eknæs M, Lind GE, Myklebost O, Skotheim RI, Sveen A, Lothe RA, 2017. Multi-omics of 34 colorectal cancer cell lines-a resource for biomedical studies. Mol Cancer, 16(1): 116.
Casañas-Sánchez V, Pérez JA, Quinto-Alemany D, Díaz M, 2016. Sub-toxic ethanol exposure modulates gene expression and enzyme activity of antioxidant systems to provide neuroprotection in hippocampal HT22 cells. Front Physiol, 7: 312.
Clave S, Joya X, Salat-Batlle J, Garcia-Algar O, Vall O, 2014. Ethanol cytotoxic effect on trophoblast cells. Toxicol Lett, 225(2): 216-221.
Da Violante G, Zerrouk N, Richard I, Provot G, Chaumeil JC, Arnaud P, 2002. Evaluation of the cytotoxicity effect of dimethyl sulfoxide (DMSO) on Caco2/TC7 colon tumor cell cultures. Biol Pharm Bull, 25(12): 1600-1603.
de Abreu Costa L, Henrique Fernandes Ottoni M, Dos Santos MG, Meireles AB, Gomes de Almeida V, de Fátima Pereira W, Alves de Avelar-Freitas B, Eustáquio Alvim Brito- Melo G, 2017. Dimethyl sulfoxide (DMSO) decreases cell proliferation and TNF-α, IFN-γ, and IL-2 cytokines production in cultures of peripheral blood lymphocytes. Molecules, 22(11): E1789.
Demir S, Turan I, Aliyazicioglu Y, 2018a. Cytotoxic effect of Rhododendron luteum leaf extract on human cancer cell lines. KSU J Agric Nat, 21(6): 950-956.
Demir S, Turan I, Aliyazicioglu R, Ozer Yaman S, Aliyazicioglu Y, 2018b. Primula vulgaris extract induces cell cycle arrest and apoptosis in human cervix cancer cells. J Pharm Anal, 8(5): 307-311.
Demir S, Turan I, Aliyazicioglu Y, 2019a. Antioxidant properties of Primula vulgaris flower extract and its cytotoxic effect on human cancer cell lines. KSU J Agric Nat, 22(1): 78-84.
Demir S, Turan I, Misir S, Aliyazicioglu Y, 2019b. Selective cytotoxic effect of Dorycnium pentaphyllum extract on human breast, liver, and lung cancer cells. KSU J Agric Nat, 22(3): 473-479.
Demir S, Yaman SO, Sener SO, Demir EA, Aliyazicioglu R, Ozgen U, Mentese A, Deger O, Aliyazicioglu Y, 2020. Dorycnium pentaphyllum extract has antiproliferative effect on human cervix and colon cancer cells. Nutr Cancer, 72(3): 504-512.
Ferreira D, Silva AP, Nobrega FL, Martins IM, Barbosa-Matos C, Granja S, Martins SF, Baltazar F, Rodrigues LR, 2019. Rational identification of a colorectal cancer targeting peptide through phage display. Sci Rep, 9: 3958.
Forman S, Kas J, Fini F, Steinberg M, Ruml T, 1999. The effect of different solvents on the ATP/ADP content and growth properties of HeLa cells. J Biochem Mol Toxicol, 13(1): 11-15.
Galvao J, Davis B, Tilley M, Normando E, Duchen MR, Cordeiro MF, 2014. Unexpected low-dose toxicity of the universal solvent DMSO. FASEB J, 28(3): 1317-1330.
Gazdar AF, Girard L, Lockwood WW, Lam WL, Minna JD, 2010. Lung cancer cell lines as tools for biomedical discovery and research. J Natl Cancer Inst, 102(17): 1310-1321.
Hajighasemi F, Tajik S, 2017. Assessment of cytotoxicity of dimethyl sulfoxide in human hematopoietic tumor cell lines. IJBC, 9(2): 48-53.
Hebling J, Bianchi L, Basso FG, Scheffel DL, Soares DG, Carrilho MRO, Pashley DH, Tjäderhane L, de Souza Costa CA, 2015. Cytotoxicity of dimethyl sulfoxide (DMSO) in direct contact with odontoblast-like cells. Dent Mater, 31(4): 399-405.
Jamalzadeh L, Ghafoori H, Sariri R, Rabuti H, Nasirzade J, Hasani H, Aghamaali MR, 2016. Cytotoxic effects of some common organic solvents on MCF-7, RAW-264.7 and human umbilical vein endothelial cells. Avicenna J Med Biochem, 4(1): e33453.
Kade S, Herzog N, Schmidtke KU, Küpper JH, 2016. Chronic ethanol treatment depletes glutathione regeneration capacity in hepatoma cell line HepG2. J Cell Biotech, 1(2): 183-190.
Liu S, Gao G, Yan D, Chen X, Yao X, Guo S, Li G, Zhao Y, 2017. Effects of miR-145-5p through NRAS on the cell proliferation, apoptosis, migration, and invasion in melanoma by inhibiting MAPK and PI3K/AKT pathways. Cancer Med, 6(4): 819-833.
Mikami K, Haseba T, Ohno Y, 1997. Ethanol induces transient arrest of cell division (G2+M block) followed by G0/G1 block: dose effects of shortand longer-term ethanol exposure on cell cycle and cell functions. Alcohol Alcohol, 32(2): 145-152.
Misir S, Aliyazicioglu Y, Demir S, Turan I, Yaman SO, Deger O, 2018. Antioxidant properties and protective effect of Turkish propolis on t-BHP-induced oxidative stress in foreskin fibroblast cells. IJPER, 52(1): 94-100.
Misir S, Aliyazicioglu Y, Demir S, Turan I, Hepokur C, 2020. Effect of Turkish propolis on miRNA expression, cell cycle, and apoptosis in human breast cancer (MCF-7) cells. Nutr Cancer, 72(1):133-145.
Mosmann T, 1983. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods, 65: 55-63.
Schäuble S, Klement K, Marthandan S, Münch S, Heiland I, Schuster S, Hemmerich P, Diekmann S, 2012. Quantitative model of cell cycle arrest and cellular senescence in primary human fibroblasts. PLoS One, 7(8): e42150.
Singh M, McKenzie K, Ma X, 2017. Effect of dimethyl sulfoxide on in vitro proliferation of skin fibroblast cells. J Biotech Res, 8: 78-82.
Timm M, Saaby L, Moesby L, Hansen EW, 2013. Considerations regarding use of solvents in in vitro cell based assays. Cytotechnology, 65: 887-894.
Turan I, Demir S, Kilinc K, Aliyazicioglu Y, Alver A, Misir S, Ozer Yaman S, Akbulut K, Mentese A, Deger O, 2017. Morus rubra extract induces G1 cell cycle arrest and apoptosis in human lung and prostate cancer cells. IJPER, 51(1): 51-58.
Turan I, Demir S, Kilinc K, Yaman SO, Misir S, Kara H, Genc B, Mentese A, Aliyazicioglu Y, Deger O, 2018. Cytotoxic effect of Rosa canina extract on human colon cancer cells through repression of telomerase expression. J Pharm Anal, 8(6): 394-399.
Wu D, Bahaerguli K, Wu GR, Sheng L, 2010. Studies on the effect of DMSO as a solvent mediator on cell growth and viability. J Xinjiang Med Univ, 33: 489-491.
Yeo EJ, Lim HK, Park SC, 2000. Effect of short-term ethanol on the proliferative response of Swiss 3T3 cells to mitogenic growth factors. Exp Mol Med, 32(3): 161-169.
Yi X, Liu M, Luo Q, Zhuo H, Cao H, Wang J, Han Y, 2017. Toxic effects of dimethyl sulfoxide on red blood cells, platelets, and vascular endothelial cells in vitro. FEBS Open Bio, 7: 485-494.
Yuan C, Gao J, Guo J, Bai L, Marshall C, Cai Z, Wang L, Xiao M, 2014. Dimethyl sulfoxide damages mitochondrial integrity and membrane potential in cultured astrocytes. PLoS One, 9(9): e107447

There are 35 citations in total.

Details

Primary Language	English
Journal Section	RESEARCH ARTICLE
Authors	Elif Ayazoglu Demir 0000-0001-9027-7633 Selim Demir 0000-0002-1863-6280 Yüksel Aliyazıcıoğlu 0000-0001-9474-4307
Publication Date	October 31, 2020
Submission Date	March 12, 2020
Acceptance Date	April 24, 2020
Published in Issue	Year 2020Volume: 23 Issue: 5

Cite

APA	Ayazoglu Demir, E., Demir, S., & Aliyazıcıoğlu, Y. (2020). In vitro Cytotoxic Effect of Ethanol and Dimethyl Sulfoxide on Various Human Cell Lines. Kahramanmaraş Sütçü İmam Üniversitesi Tarım Ve Doğa Dergisi, 23(5), 1119-1124. https://doi.org/10.18016/ksutarimdoga.vi.702702