大黄素对顺铂耐药卵巢癌细胞的逆转作用及其相关基因表达研究

The Reversal Effect of Emodin on Cisplatin Resistance in Ovarian Cancer Cells and the Expression of Resistance-associated Genes

为了研究大黄素对人卵巢癌耐药细胞株SKOV3/DDP细胞耐药逆转作用及其机制,本研究以卵巢癌SK-OV3和多药耐药细胞株SKOV3/DDP为研究对象,通过噻唑蓝(MTT)法测定SKOV3/DDP细胞的耐药指数和大黄素在无细胞毒浓度下对卵巢癌细胞耐受顺铂(DDP)的逆转作用;采用Real-time PCR技术检测耐药相关基因HIF-1α、STAT1、CK2α、GSTP1 mRNA表达情况。结果发现无细胞毒作用浓度的7.8 mg/L和3.9 mg/L大黄素能逆转SKOV3/DDP细胞对DDP的耐药性,对DDP的逆转倍数分别为1.91倍和1.30倍,与SKOV3比较,SKOV3/DDP细胞的HIF-1α、STAT1、CK2α、GSTP1 mRNA表达明显升高(P<0.01)。3.9 mg/L和7.8 mg/L大黄素作用均可下调HIF-1α、CK2α、STAT1 mRNA表达,存在剂量-效应依赖关系。7.8 mg/L大黄素作用可下调GSTP1 mR-NA表达,但3.9 mg/L大黄素作用不明显。7.8 mg/L和3.9 mg/L大黄素联合IC50浓度的DDP时,四个耐药相关基因的表达与单独化疗药组作用相比明显下调(P<0.01)。提示无细胞毒浓度的大黄素对卵巢癌细胞耐药逆转的作用可能是通过下调HIF-1α、STAT1、GSTP1、CK2α的表达起作用。

To inversitigate the drug resistance reversal effect and mechanism of emodin in cisplatinum-resistant ovarian cancer cells SKOV3/DDP（short for cisplatin） in vitro and study the expression of HIF-1α,STAT1,and GSTP1 resistance-associated genes.Methyl thiazolyl tetrazolium（MTT） method was used to detect the reversal index（RI） of SKOV3/DDP cells and the reversal effect of emodin.The resistance-associated genes expression of HIF-1α,CK2α,STAT1,and GSTP1 was detected by Real time PCR.The results showed that noncytotoxic dose of emodin reversed the cisplatin-resistance of SKOV-3/DDP,the reversal index（RI） was 1.91 and 1.30 respectively for 7.8 mg/L and 3.9 mg/L emodin.Compared with SKOV3 cells,expression level of HIF-1α,STAT1,CK2α,and GSTP1 mRNA in SKOV3/DDP was significantly raised（P0.01）.3.9 mg/L and 7.8 mg/L emodin could down regulated the gene expression of HIF-1α,CK2α,and STAT1 in a dose-dependent manner.Though 7.8 mg/L emodin down-regulated GSTP1 mRNA expression,3.9 mg/L emodin had no such effects.The mRNA expression level of those gene was significantly down-regulated when 7.8 mg/L and 3.9 mg/L emodin was combined with DDP（P0.01）.Emodin can down regulate the gene expression of HIF-1α,CK2α,STAT1,and GSTP1 on SKOV-3/DDP cells,which may account for the reversal effect of emodin on cisplatin resistance in ovarian cancer cells.