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

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。

Abstract:

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<0.01); ③Compared with the control group, the VEGF protein expression decreased obviously after stimulated with ART (P<0.01); ④Same result were observed in VEGF mRNA expression by RT-qPCR (P<0.01). Conclusion: ART could induce apoptosis, decrease VEGF expression and invasion of NACC cells in vitro.