HEPG2 Hücrelerinde Apoptotik Proteinlerin ve Çoklu İlaç Direnci Geninin Ekspresyon Düzeyleri

Year 2012, Issue: 2, 119 - 132, 01.06.2012

Gözde Aydın Ayşe Ercan

Abstract

Bu çalışmada, kanser hücrelerinde işleyişi hasarlı olan apoptotik, anti-apoptotik mekanizmaların ve çoklu ilaç direnci yolakların incelenmesi amacıyla, karaciğer kanseri hücre hattı HepG2 kullanılmıştır. Bu çalışmanın amacı: HepG2 hücre hattını en etkili antikanser ilaçlardan biri olan Doksorubisin ile muamele ederek, apoptozun uyarılması ve apoptozda önemli rol oynayan birbiriyle ilişkili p53, Bcl-2 ve MDR-1 genlerinin ekspresyon düzeylerinin incelenmesidir. Bu amaçla hücreler ilaç ile muamele edilerek büyütülmüş, sonrasında RNA’ları saflaştırılmış, cDNA’ya dönüştürülmüş ve elde edilen cDNA’lar üç genin ekspresyon düzeyini belirlemek amacıyla RT-PCR cihazında/yönteminde kullanılmıştır. HepG2 hücrelerinde 50 nM, 200 nM, 400 nM ve 800 nM ilaç uygulanmasından sonra kontrol gruplarına kıyasla deney gruplarında, p53 geninin ekspresyon düzeylerinin sırasıyla 2,176, 1,718, 1,524, 1,225 kat arttığı, Bcl-2 gen ekspresyon düzeylerinin sırasıyla 4,182, 6,512, 5,641, 3,910 kat arttığı ve MDR-1 gen ekspresyon düzeyinin ise uygulanan ilaç dozundan bağımsız olarak değişmediği gözlenmiştir.

Keywords

Gen ekspresyonu, p53, bcl-2, mdr-1, HepG2

Gene Expression Levels of Apoptotic Proteins and Multidrug Resistance Genes in HEPG2 Cells

Abstract

In this study, liver cancer cell line HepG2 were used with the aim of examining the apoptotic-antiapoptotic mechanisms and multi drug resistance pathways which are defective in cancer cells. The aim of this study: is to trigger apoptosis by treatment HepG2 cancer cell line with one of the most effective anticancer drug Doxorubicin and to investigate the expression levels of p53, Bcl-2 and MDR-1 genes which are related with each other and play an important role in apoptosis. With this purpose, the cells were grown by treating with increasing doses of Doxorubicin. Their RNA’s were purified, converted into cDNAs and used in Real Time PCR device in order to determine the expression levels of these genes. It is observed in HepG2 cells, compared to the control groups after 50 nM, 200 nM, 400 nM ve 800 nM drug treatment, p53 gene expression level is respectively 2,176, 1,718, 1,524, 1,225 fold increased, Bcl-2 gene expression level is respectively 4,182, 6,512, 5,641, 3,910 fold increased and MDR-1 gene expression level is not dependent to the treated drug dose.

Keywords

Gene expression, p53, bcl-2, mdr-1, HepG2

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