The role of interactions of CYP1A1, HBX and NF-kB in the development of hepatocelular carcinoma

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Tezin Türü: Yüksek Lisans

Tezin Yürütüldüğü Kurum: Pamukkale Üniversitesi, Fen-Edebiyat Fakültesi, Biyoloji Bölümü, Türkiye

Tezin Onay Tarihi: 2013

Tezin Dili: Türkçe

Öğrenci: Tuğba Koç

Asıl Danışman (Eş Danışmanlı Tezler İçin): Alaattin Şen

Eş Danışman: Naciye Lale Tufan

Özet:

In this study, the potential interactions among Hepatitis BX (HBX), Cytochrome P450 1A1 (CYP1A1) and Nuclear Factor Kappa B (NF-?B) proteins in Hepatitis B virus mediated human liver cancer cell line (HepG2) were studied using molecular approaches. For this purpose, CYP1A1 and NF-kB (RelA) genes were transiently transfected into HepG2 cells. After transfection, HBX, CYP1A1 and RelA levels were detected by measuring the related mRNA expressions using Reverse Transcriptase Polymerase Change Reaction (RT-PCR). In addition, CYP1A1 protein level was also determined by measuring CYP1A-directed EROD activity and western blot using specific anibodies against human CYP1A1. Similarly, RelA expressions were investigated by westren blotting. Furthermore, the effect of CYP1A1 and RelA expressions on the survival of HepG2 cells were monitored using cytotoxicity assays. Finally, the protein-protein interactions between CYP1A1 and HBX was determined by co-immunoprecipitation analysis. The transfection efficiency of CYP1A1 and RelA genes in HepG2 cells were observed between 20%-40%. The level mRNA expression of CYP1A1 and RelA genes and EROD activity further confirmed the low level of tranfection efficiencies. CYP1A1 and RelA did not show any toxic effects on the survival of the HepG2 cells. Western blot studies demonstrated the low level expression of CYP1A1 protein in these cells. Co-immunoprecipitation analysis clearly depicted that there is no protein-protein interaction between CYP1A1 and HBX in the HepG2 cells. Therefore, HBX CYP1A1 proteins do not interact with each other in HBX transfected human liver cancer cells. Consequently CYP1A1 gene may not play a role in the mechanism of HBV. Key words: Hepatit B X protein, Cytohrome P450 1A1, Nuclear Factor Kappa B