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

Authors : Mustafa SONGUR, Sema BİLGİN, Seçil ERDEN TAYHAN, İsa GÖKÇE

Pages : 2943-2951

Doi:10.21597/jist.730267

View : 22 | Download : 7

Publication Date : 2020-12-15

Article Type : Research Paper

Abstract :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.
Keywords : Recombinant J2 C2, therapeutic protein, cell proliferation, cell migration, apoptosis