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作 者:俞立平[1] 农晓琳[1] 李佳荃[2] 胥航[1] 朴智[1]
机构地区:[1]广西医科大学口腔医学院口腔颌面外科,南宁530021 [2]广西医科大学医学科学实验中心,南宁530021
出 处:《中国实验方剂学杂志》2013年第9期237-241,共5页Chinese Journal of Experimental Traditional Medical Formulae
基 金:国家自然科学基金项目(30060082);教育部"春晖计划"科研启动基金(教外司留[2003]593-7);广西卫生厅重点科研项目(重200006;重200927);广西科学基金项目(2010GXNSFD013047;桂科回0836013)
摘 要:目的:探究青蒿琥酯(ART)对人腭部小涎腺腺样囊性癌(NACC)细胞凋亡、侵袭能力的影响。方法:NACC细胞经ART 100 mg.L-1干预后,吖啶橙/溴化乙锭(AO/EB)染色观察细胞形态学变化;Transwell小室检测对照组及ART 25,50,100 mg.L-1组细胞侵袭力;免疫细胞化学法测定各组细胞胞浆中血管内皮生长因子(VEGF)的蛋白表达量;实时荧光定量聚合酶链反应(Real-time qPCR)测定各组细胞VEGF mRNA的表达。结果:①经AO/EB染色,与对照组相比,ART 100 mg.L-1组可明显观察到凋亡细胞(细胞形态不规则,核染色质固缩);②Transwell实验中,与对照组相比,ART 25,50,100 mg.L-1组细胞跨膜数减少(P<0.01);③细胞免疫化学结果显示,与对照组相比,ART 25,50,100,200 mg.L-1各组胞浆中VEGF的蛋白阳性表达量显著降低(P<0.01);④Real-time qPCR测得各ART组VEGF基因相对表达量均明显低于对照组(P<0.01)。结论:青蒿琥酯可诱导NACC细胞的凋亡,抑制其侵袭能力,下调其VEGF的表达。Objective:To investigate the influence of artesunate(ART) on invasion and apoptosis of novel human palatal salivary gland cell line of adenoid cystic carcinoma(NACC).Method: After treated with ART 100 mg · L-1,apoptosis cells were observed after loaded with acridine orange/ethidium bromide dye(AO/EB).The anti-invasion effect of ART at 25,50,100 mg · L-1 were examined by transwell assay.The expression of vascular endothelial growth factor(VEGF) and its mRNA of NACC cells was evaluated by immunohistochemistry and RT-qPCR technique separately.Result:①Compared with the control group,apoptosis cells were observed after treated with ART 100 mg · L-1,with irregular and shrinked nucleolus;②The result of transwell assay showed that ART at 25,50,100 mg · L-1 could inhibit the transmembrane cells obviously compared with control group(P&lt;0.01);③Compared with the control group,the VEGF protein expression decreased obviously after stimulated with ART(P&lt;0.01);④Same result were observed in VEGF mRNA expression by RT-qPCR(P&lt;0.01).Conclusion: ART could induce apoptosis,decrease VEGF expression and invasion of NACC cells in vitro.
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