Araştırma Makalesi
BibTex RIS Kaynak Göster

Investigation of Anti-proliferative and Anti-migratory Properties of Recombinant J2-C2 Against Tumor Cells

Yıl 2020, Cilt: 10 Sayı: 4, 2943 - 2951, 15.12.2020
https://doi.org/10.21597/jist.730267

Öz

Cancer is a big public health problem in many parts of the world. A novel anti-tumor protein (J2-C2) was previously isolated from Arca inflata and it was reported that this protein had anti-proliferative effect on some human tumor cell lines such as A549, HepG2 and SPC-A-1. In this study, firstly, J2-C2 was produced by recombinant techniques in the Escherichia coli strain BL21 (DE3) pLysE and this protein was purified by Ni-NTA agarose affinity chromatography. Expressed recombinant J2-C2 was analyzed with SDS-PAGE. 75.5 mg ml-1 of J2-C2 was achieved from a 600 mL culture. Then using HT-29, MCF7 and PC3 cancer cell lines, we showed the effect of recombinant of J2-C2 on cell proliferation, migration and apoptosis in a cell specific manner. Cell viability was measured using MTT assay. Additionally, real-time-qPCR was applied to analyze the transcript levels of apoptosis related genes such as Bcl-2, Bax and p53. The 2–ΔΔCt method was performed to determine the relative changes in gene transcription. Moreover, scratch wound healing assay was performed to evaluate the effect of J2-C2 on cancer cell migration. Consequently, we found that recombinant J2-C2 did not have a significant effect on cell viabilities of MCF7, PC3 and HT29 in concentration-dependent manner. Furthermore, our results showed that recombinant J2-C2 declined HT29, MCF7 cell migration. However, we did not observe the same results for PC3 cancer cell line.

Destekleyen Kurum

TUBİTAK-BIDEB 2209B

Kaynakça

  • Andersen DC, Krummen L, 2002. Recombinant protein expression for therapeutic applications. Current Opinion in Biotechnology. 13 (2), 117–123.
  • Assenberg R, Wan PT, Geisse S, Mayr, LM, 2013. Advances in recombinant protein expression for use in pharmaceutical research. Current Opinion in Structural Biology, 23 (3), 393-402.
  • Cabrita LD, Chow MKM, Bottomley SP, 2004. A Practical Guide To Protein Expression and Refolding From Inclusion Bodies. Biotechnolgy Annual Review, 10, 31–50.
  • Cheok C F, 2012. Protecting normal cells from the cytotoxicity of chemotherapy. Cell Cycle 11 (12) 2227-2232.
  • Fan M, Wen Y, Ye D, Jin Z, Zhao P, Chen D, Lu X, He Q, 2019. Acid-Responsive H2-Releasing 2D MgB2 Nanosheet for Therapeutic Synergy and Side Effect Attenuation of Gastric Cancer Chemotherapy. Advanced Healthcare Materials. 8 (13).
  • Qian CN, Mei Y, Zhang J, 2017. Cancer metastasis: issues and challenges. Chinese Journal of Cancer, 36(38).
  • Ispir E, İkiz M, İnan A, Sünbül, AB, Erden Tayhan S, Bilgin S, Elmastaş M, 2019. Synthesis, structural characterization, electrochemical, photoluminescence, antiproliferative and antioxidant properties of Co(II), Cu(II) and Zn(II) complexes bearing the azo-azomethine ligands. Journal of Molecular Structure, 1182, 63–71 (2019).
  • Inan A, Sünbül AB, İkiz B, Erden Tayhan S, Bilgin S, Elmastaş M, Sayın K, Ceyhan G, Köse M, İspir E, 2018. Half-sandwich Ruthenium(II) arene complexes bearing the azo-azomethine ligands: Electrochemical, computational, antiproliferative and antioxidant properties. Journal of Organometallic Chemistry, 870, 76–89.
  • Li H, Su J, Jiang J, Li Y, Gan Z, Ding Y, Li Y, Liu J, Wang S, Ke Y, 2019. Characterization of polysaccharide from Scutellaria barbata and its antagonistic effect on the migration and invasion of HT-29 colorectal cancer cells induced by TGF-β1. International Journal of Biological Macromolecules, 131, 886-895.
  • Li J J, Li Q, 2008. Isolation and characterization of twelve novel microsatellite loci in the ark shell Scapharca broughtonii.Conserv. Genet., 9, 1055-1057.
  • Ortega A, 2003. A new role for GABA: inhibition of tumor cell migration. Trends in Pharmacological Sciences, 24 (4), 151-154.
  • Priego S, Feddi F, Ferrer P, Mena S, Benlloch M, Ortega A, Carretero J, Obrador E,Asensi M,and Estrela JM, 2008. Natural polyphenols facilitate elimination of HT-29 colorectalcancer xenografts by chemoradiotherapy: a Bcl-2- and superoxide dismutase 2-dependent mechanism. Molecular Cancer Therapeutics, 7(10).
  • Rao X, Huang X, Zhou Z, Lin X, 2013. An improvement of the 2ˆ(–delta delta CT) method for quantitative real-time polymerase chain reaction data analysis. Biostat Bioinforma Biomath. 3(3), 71–85.
  • Rosano GL, Ceccarelli EA, 2014. Recombinant protein expression in Escherichia coli: Advances and challenges. Frontier in Microbiology, 5, 1–17.
  • Saito Y, Kitagawa W, Kumaga T, Tajima N, Nishimiya Y, Tamano K, Yasutake Y, Tamura T, Kameda T, 2019. Developing a codon optimization method for improved expression of recombinant proteins in actinobacteria scientific reports. Scientific Reports. 9:8338.
  • Swiech K, Picanço-Castro V, Covas T, Covas DT, 2012. Human cells: New platform for recombinant therapeutic protein production. Protein Expression and Purification. 84, 147–153.
  • Weber K, Pringle J, Osborn M, 1972. Measurement of molecular weights by electrophoresis on SDS-acrylamide gel. Methods in Enzymology, 26, 3–27.
  • Xu J, Chen Z, Song L, Chen L, Zhu J, Lv S, Yu R, 2013. A New in Vitro Anti-Tumor Polypeptide Isolated from Arca inflata. Marine Drugs, 11 (12), 4773.
  • Zhu P, Zhao N, Sheng D, Hou J, Hao C, Yang X, Zhu B, Zhang S, Han Z, Wei L, Zhang L, 2016. Inhibition of Growth and Metastasis of Colon Cancer by Delivering 5-Fluorouracil-loaded Pluronic P85 Copolymer Micelles. Scientific Reports, 6, 1-11.
  • Zhu J, Xu J, Wang Y, Li C, Chen Z, Song L, Gao J, Yu R, 2017. Purification and structural characterization of anti-tumor protein from Arca inflata. International Journal of Biological Macromolecules. 105, 103–110.

Investigation of Anti-proliferative and Anti-migratory Properties of Recombinant J2-C2 Against Tumor Cells

Yıl 2020, Cilt: 10 Sayı: 4, 2943 - 2951, 15.12.2020
https://doi.org/10.21597/jist.730267

Öz

Cancer is a big public health problem in many parts of the world. A novel anti-tumor protein (J2-C2) was previously isolated from Arca inflata and it was reported that this protein had anti-proliferative effect on some human tumor cell lines such as A549, HepG2 and SPC-A-1. In this study, firstly, J2-C2 was produced by recombinant techniques in the Escherichia coli strain BL21 (DE3) pLysE and this protein was purified by Ni-NTA agarose affinity chromatography. Expressed recombinant J2-C2 was analyzed with SDS-PAGE. 75.5 mg ml-1 of J2-C2 was achieved from a 600 mL culture. Then using HT-29, MCF7 and PC3 cancer cell lines, we showed the effect of recombinant of J2-C2 on cell proliferation, migration and apoptosis in a cell specific manner. Cell viability was measured using MTT assay. Additionally, real-time-qPCR was applied to analyze the transcript levels of apoptosis related genes such as Bcl-2, Bax and p53. The 2–ΔΔCt method was performed to determine the relative changes in gene transcription. Moreover, scratch wound healing assay was performed to evaluate the effect of J2-C2 on cancer cell migration. Consequently, we found that recombinant J2-C2 did not have a significant effect on cell viabilities of MCF7, PC3 and HT29 in concentration-dependent manner. Furthermore, our results showed that recombinant J2-C2 declined HT29, MCF7 cell migration. However, we did not observe the same results for PC3 cancer cell line.

Kaynakça

  • Andersen DC, Krummen L, 2002. Recombinant protein expression for therapeutic applications. Current Opinion in Biotechnology. 13 (2), 117–123.
  • Assenberg R, Wan PT, Geisse S, Mayr, LM, 2013. Advances in recombinant protein expression for use in pharmaceutical research. Current Opinion in Structural Biology, 23 (3), 393-402.
  • Cabrita LD, Chow MKM, Bottomley SP, 2004. A Practical Guide To Protein Expression and Refolding From Inclusion Bodies. Biotechnolgy Annual Review, 10, 31–50.
  • Cheok C F, 2012. Protecting normal cells from the cytotoxicity of chemotherapy. Cell Cycle 11 (12) 2227-2232.
  • Fan M, Wen Y, Ye D, Jin Z, Zhao P, Chen D, Lu X, He Q, 2019. Acid-Responsive H2-Releasing 2D MgB2 Nanosheet for Therapeutic Synergy and Side Effect Attenuation of Gastric Cancer Chemotherapy. Advanced Healthcare Materials. 8 (13).
  • Qian CN, Mei Y, Zhang J, 2017. Cancer metastasis: issues and challenges. Chinese Journal of Cancer, 36(38).
  • Ispir E, İkiz M, İnan A, Sünbül, AB, Erden Tayhan S, Bilgin S, Elmastaş M, 2019. Synthesis, structural characterization, electrochemical, photoluminescence, antiproliferative and antioxidant properties of Co(II), Cu(II) and Zn(II) complexes bearing the azo-azomethine ligands. Journal of Molecular Structure, 1182, 63–71 (2019).
  • Inan A, Sünbül AB, İkiz B, Erden Tayhan S, Bilgin S, Elmastaş M, Sayın K, Ceyhan G, Köse M, İspir E, 2018. Half-sandwich Ruthenium(II) arene complexes bearing the azo-azomethine ligands: Electrochemical, computational, antiproliferative and antioxidant properties. Journal of Organometallic Chemistry, 870, 76–89.
  • Li H, Su J, Jiang J, Li Y, Gan Z, Ding Y, Li Y, Liu J, Wang S, Ke Y, 2019. Characterization of polysaccharide from Scutellaria barbata and its antagonistic effect on the migration and invasion of HT-29 colorectal cancer cells induced by TGF-β1. International Journal of Biological Macromolecules, 131, 886-895.
  • Li J J, Li Q, 2008. Isolation and characterization of twelve novel microsatellite loci in the ark shell Scapharca broughtonii.Conserv. Genet., 9, 1055-1057.
  • Ortega A, 2003. A new role for GABA: inhibition of tumor cell migration. Trends in Pharmacological Sciences, 24 (4), 151-154.
  • Priego S, Feddi F, Ferrer P, Mena S, Benlloch M, Ortega A, Carretero J, Obrador E,Asensi M,and Estrela JM, 2008. Natural polyphenols facilitate elimination of HT-29 colorectalcancer xenografts by chemoradiotherapy: a Bcl-2- and superoxide dismutase 2-dependent mechanism. Molecular Cancer Therapeutics, 7(10).
  • Rao X, Huang X, Zhou Z, Lin X, 2013. An improvement of the 2ˆ(–delta delta CT) method for quantitative real-time polymerase chain reaction data analysis. Biostat Bioinforma Biomath. 3(3), 71–85.
  • Rosano GL, Ceccarelli EA, 2014. Recombinant protein expression in Escherichia coli: Advances and challenges. Frontier in Microbiology, 5, 1–17.
  • Saito Y, Kitagawa W, Kumaga T, Tajima N, Nishimiya Y, Tamano K, Yasutake Y, Tamura T, Kameda T, 2019. Developing a codon optimization method for improved expression of recombinant proteins in actinobacteria scientific reports. Scientific Reports. 9:8338.
  • Swiech K, Picanço-Castro V, Covas T, Covas DT, 2012. Human cells: New platform for recombinant therapeutic protein production. Protein Expression and Purification. 84, 147–153.
  • Weber K, Pringle J, Osborn M, 1972. Measurement of molecular weights by electrophoresis on SDS-acrylamide gel. Methods in Enzymology, 26, 3–27.
  • Xu J, Chen Z, Song L, Chen L, Zhu J, Lv S, Yu R, 2013. A New in Vitro Anti-Tumor Polypeptide Isolated from Arca inflata. Marine Drugs, 11 (12), 4773.
  • Zhu P, Zhao N, Sheng D, Hou J, Hao C, Yang X, Zhu B, Zhang S, Han Z, Wei L, Zhang L, 2016. Inhibition of Growth and Metastasis of Colon Cancer by Delivering 5-Fluorouracil-loaded Pluronic P85 Copolymer Micelles. Scientific Reports, 6, 1-11.
  • Zhu J, Xu J, Wang Y, Li C, Chen Z, Song L, Gao J, Yu R, 2017. Purification and structural characterization of anti-tumor protein from Arca inflata. International Journal of Biological Macromolecules. 105, 103–110.
Toplam 20 adet kaynakça vardır.

Ayrıntılar

Birincil Dil İngilizce
Bölüm Moleküler Biyoloji ve Genetik / Moleculer Biology and Genetic
Yazarlar

Mustafa Songur 0000-0002-8581-9992

Sema Bilgin

Seçil Erden Tayhan

İsa Gökçe 0000-0002-5023-9947

Yayımlanma Tarihi 15 Aralık 2020
Gönderilme Tarihi 1 Mayıs 2020
Kabul Tarihi 19 Temmuz 2020
Yayımlandığı Sayı Yıl 2020 Cilt: 10 Sayı: 4

Kaynak Göster

APA Songur, M., Bilgin, S., Erden Tayhan, S., Gökçe, İ. (2020). Investigation of Anti-proliferative and Anti-migratory Properties of Recombinant J2-C2 Against Tumor Cells. Journal of the Institute of Science and Technology, 10(4), 2943-2951. https://doi.org/10.21597/jist.730267
AMA Songur M, Bilgin S, Erden Tayhan S, Gökçe İ. Investigation of Anti-proliferative and Anti-migratory Properties of Recombinant J2-C2 Against Tumor Cells. Iğdır Üniv. Fen Bil Enst. Der. Aralık 2020;10(4):2943-2951. doi:10.21597/jist.730267
Chicago Songur, Mustafa, Sema Bilgin, Seçil Erden Tayhan, ve İsa Gökçe. “Investigation of Anti-Proliferative and Anti-Migratory Properties of Recombinant J2-C2 Against Tumor Cells”. Journal of the Institute of Science and Technology 10, sy. 4 (Aralık 2020): 2943-51. https://doi.org/10.21597/jist.730267.
EndNote Songur M, Bilgin S, Erden Tayhan S, Gökçe İ (01 Aralık 2020) Investigation of Anti-proliferative and Anti-migratory Properties of Recombinant J2-C2 Against Tumor Cells. Journal of the Institute of Science and Technology 10 4 2943–2951.
IEEE M. Songur, S. Bilgin, S. Erden Tayhan, ve İ. Gökçe, “Investigation of Anti-proliferative and Anti-migratory Properties of Recombinant J2-C2 Against Tumor Cells”, Iğdır Üniv. Fen Bil Enst. Der., c. 10, sy. 4, ss. 2943–2951, 2020, doi: 10.21597/jist.730267.
ISNAD Songur, Mustafa vd. “Investigation of Anti-Proliferative and Anti-Migratory Properties of Recombinant J2-C2 Against Tumor Cells”. Journal of the Institute of Science and Technology 10/4 (Aralık 2020), 2943-2951. https://doi.org/10.21597/jist.730267.
JAMA Songur M, Bilgin S, Erden Tayhan S, Gökçe İ. Investigation of Anti-proliferative and Anti-migratory Properties of Recombinant J2-C2 Against Tumor Cells. Iğdır Üniv. Fen Bil Enst. Der. 2020;10:2943–2951.
MLA Songur, Mustafa vd. “Investigation of Anti-Proliferative and Anti-Migratory Properties of Recombinant J2-C2 Against Tumor Cells”. Journal of the Institute of Science and Technology, c. 10, sy. 4, 2020, ss. 2943-51, doi:10.21597/jist.730267.
Vancouver Songur M, Bilgin S, Erden Tayhan S, Gökçe İ. Investigation of Anti-proliferative and Anti-migratory Properties of Recombinant J2-C2 Against Tumor Cells. Iğdır Üniv. Fen Bil Enst. Der. 2020;10(4):2943-51.