青蒿琥酯对腺样囊性癌NACC细胞凋亡和侵袭的影响

Effect of Artesunate on Apoptosis and Invasion of Novel Human Palatal Salivary Gland Cell Line of Adenoid Cystic Carcinoma in vitro

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.