Effect of Carthami Tinctorii Fructus Herbal-acupuncture Solution(CTF-HAS) on Gene Expression in HepG2 carcinomar cells

Oligonucleotide chip를 이용한 홍화자약침액(紅花子藥鍼液)이 간암세포주(肝癌細胞柱)의 유전자(遺傳子) 발현(發顯)에 미치는 영향(影響)

Objective : It has long been known about the osteogenic effect of CTF-HAS on bone tissues. However, it has not been determined the effect of CTF-HAS on cancer cells. The purpose of this study is to screen the CTF-HAS mediated differentially expressed genes in cancer cells such as HepG2 hepatoma cells lines. Oligonucleotide microarray approach were employed to screen the differential expression genes. Methods : CTF-HAS was prepared by boiling and stored at $-70^{\circ}C$ until use. Cells were treated with various concentrations of CTF-HAS(0.1, 0.5, 1.5, 10, $20mg/m{\ell}$) for 24 h. Cytotoxicity was tested by MTT assay. To screen the differentially expressed genes in cancer cells, cells were treated with $1.5mg/m{\ell}$ of CTF-HAS. For oligonucleotide microarray assay, total RNA was used for gene expression analysis using oligonucleotide genechip (Human genome U133 Plus 2.0., Affimatrix Co.). ResuIts : It has no cytotoxic effects on HepG2 cells in all concentrations (0.1, 0.5, 1.5, 10, $20mg/m{\ell}$). More than twofold up-regulated genes were 19 genes. The number of more than twofold down-regulated genes was 13. Discussion : This study showed the screening of CTF-HAS mediated differentially regulated genes using combined approaches of oligonucleotide microarray. The screened genes will be used for the better understanding in therapeutic effect of CTF-HAS on cancer field.

홍화자약침액(紅花子藥鍼液)의 항암효능(抗癌效能)을 밝히고자 간암세포주(肝癌細胞柱)에 최신 oligonucleotide chip assay 법을 통하여 대양(大量)의 유전자(遺傳子) 발현(發顯)을 분석(分析)한 결과(結果) 다음과 같은 결론(結論)을 얻었다. 1. MTT 분석(分析)에서 간암(肝癌) 및 위암세포주(胃癌細胞柱)는 홍화자약침액(紅花子藥鍼液) 1.5, 10, 20m/$m{\ell}$에서 대조군(對照群)에 비해 유의(有意)한 세포활성(細胞活性) 감소(減少)를 보였다. 2. 간암세포주(肝癌細胞柱)에 홍화자약침액(紅花子藥鍼液) 처치(處置) 시(時) 대조군(對照群)에 비해 발현(發顯)이 2배 이상 증가(增加)된 유전자(遺傳子)는 UCIA PMARARA fusion protein, human oral cancer candidate gene mRNA, EPPIN-3 등 19개였다. 3. 간암세포주(肝癌細胞柱)에 홍화자약침액(紅花子藥鍼液) 처치(處置) 시 (時)대조군(對照群)에 비해 유전자(遺傳子)의 발현(發顯)이 2배 이상 감소(減少)된 것은 BTF3, MMP11, paxillin, villinn 등 13개였다. 이상과 같이 홍화자약침액(紅花子藥鍼液)에 대한 간암세포주(肝癌細胞柱)의 유전자(遺傳子) 발현(發顯)을 oligonucleotide 분석(分析)으로 대량(大量) 검소(儉素)할 수 있었고, 심한 발현(發顯) 차이(差異)를 나타내는 각 유전자(遺傳子)는 암화(癌化) 과정(過程)이나, 홍화자약침액(紅花子藥鍼液)에 반응하는 유전자(遺傳子)로 치료제 개발을 위한 기초 자료로 활용할 수 있을 것으로 사료(思料)된다.