摘要
背景:人脱嘌呤脱嘧啶核酸内切酶/氧化还原因子-1(APE1/Ref-1)基因与肿瘤的化放疗抵抗和预后密切相关,是肿瘤基因治疗的理想靶点。目的:以RNA干扰技术靶向沉默人胰腺癌细胞株APE1/Ref-1基因,观察该方法对细胞增殖和凋亡的影响及其能否增强胰腺癌细胞对吉西他滨的敏感性。方法:将靶向APE1/Ref-1基因的小干扰RNA(siRNA)转染人胰腺癌细胞株SW1990,半定量逆转录聚合酶链反应(RT-PCR)和蛋白质印迹法检测APE1/Ref-1基因和蛋白表达,CCK-8检测细胞增殖情况,流式细胞仪和Hoechst 33258染色检测细胞凋亡情况。结果:转染APE1 siRNA后,SW1990细胞APE1/Ref-1 mRNA和蛋白表达显著减低,蛋白表达抑制率为55.4%±3.6%;24 h、48 h和72 h时细胞增殖抑制率分别为41.7%±2.8%、24.8%±3.7%和21.3%±9.8%;吉西他滨组、si-APE1组和联合组均可见明显细胞凋亡,联合组早期凋亡率显著高于两者单用和空白对照组(19.8%±3.5%对7.7%±1.1%、8.4%±1.0%和2.7%±1.4%,P<0.05),凋亡核形态学变化最为明显。结论:沉默APE1/Ref-1基因能抑制SW1990细胞增殖,促进细胞凋亡,显著提高细胞对吉西他滨的敏感性。RN Ai沉默APE1/Ref-1基因联合吉西他滨化疗可能成为胰腺癌治疗的新的选择。
Background： Human apurinic/apyrimidinic endonuclease/redox factor-1 （APE1/Ref-I） has been demonstrated to be associated with radioresistance, chemoresistance and poor prognosis of various cancers and might be a promising target for gene therapy of cancer. Aims： To investigate the effect of APE1/Ref-1-targeted RNA interference on proliferation and apoptosis of human pancreatic cancer cell line, and whether this modality could enhance the sensitivity of pancreatic cancer cells to gemcitabine. Methods： Small interfering RNA （siRNA） directed against APE1/Ref-1 gene was transfected into human pancreatic cancer cell line SW1990. Gene and protein expressions of APE1/Ref-1 were detected by semi-quantitative reverse transcriptase polymerase chain reaction （RT-PCR） and Western blotting, cell proliferation was assessed by cell counting kit-8 （CCK-8）, and apoptosis by flow cytometry and Hoechst 33258 staining. Restdts： After transfeeted with siRNA directed against APE1/Ref-1 gene, the protein expression of APE1/Ref-1 in SW1990 cells reduced by 55.4%±3.6%, mRNA expression also decreased significantly; the inhibition rate of cell proliferation at 24, 48 and 72 hours was 41.7%±2.8%, 24.8%＋3.7% and 21.3%＋9.8%, respectively. Apoptosis could be observed in gemcitabine group, si-APE1 group and combination group; the rate of early apoptosis was significantly higher in combination group than that in gemcitabine group, si-APE1 group and blank control group （19.8%±3.5% vs. 7.7%±1.1%, 8.4%±1.0% and 2.7%±1.4%, P〈0.05）. The apoptotic karyomorphology was more typical in combination group. Conclusions： Silencing APE1/Ref-1 gene may inhibit cell proliferation, induce apoptosis, and significantly sensitize the SW1990 ceils to gemcitabine. Therefore, gene therapy with siRNA directed against APE1/Ref-1 gene combined with gemcitabine may be a novel approach for the treatment of pancreatic cancer.
出处
《胃肠病学》
2009年第10期580-584,共5页
Chinese Journal of Gastroenterology
