氧化苦参碱对宫颈癌HeLa细胞侵袭转移的抑制作用  

Effect of oxymatrine treatment on the invasion of human cervical cancer cells

作　　者：高庆[1] 高美[1] 屈萍[2] 李岐佩[1] 陈茜[1] 

机构地区：[1]西安交通大学第二附属医院妇产科,陕西西安710004 [2]西安高新医院妇产科,陕西西安710075

出　　处：《西安交通大学学报（医学版）》2016年第3期447-450,459,共5页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：陕西省自然科学基金研究计划资助项目(No.2014JM4143)~~

摘　　要：目的观察氧化苦参碱对宫颈癌HeLa细胞侵袭转移的作用及其相关分子机制。方法 MTT法检测氧化苦参碱的抗增殖活性;Transwell小室法检测氧化苦参碱对HeLa细胞增殖的影响;Real-time PCR法检测氧化苦参碱对宫颈癌HeLa细胞侵袭转移能力的影响;Western blot法检测氧化苦参碱对MMP-2、MMP-9、AKT及p-AKT表达水平的影响。结果氧化苦参碱能够有效抑制HeLa细胞的增殖,与对照组(0mg/mL)相比,氧化苦参碱高于0.8mg/mL时出现明显差异。低于细胞毒性浓度(0.8mg/mL)的氧化苦参碱能够明显抑制HeLa细胞的侵袭能力,随着氧化苦参碱浓度的升高穿过基质胶到达小室底端的细胞数目明显减少,0.1、0.2和0.4mg/mL组分别为对照组的(77.07±20.43)%、(53.95±18.17)%、(20.35±11.20)%;0.1、0.2、0.4mg/mL组MMP-2的RNA水平分别为对照组的(82.76±8.71)%、(39.51±12.79)%、(21.53±5.38)%,0.4mg/mL组MMP-2的蛋白水平为对照组的(64.69±16.52)%;0.4mg/mL组AKT的磷酸化水平为对照组的(41.27±7.13)%。结论氧化苦参碱能够明显抑制宫颈癌HeLa细胞的侵袭转移,这种作用可能是通过调节AKT信号通路的活性进而调节MMP-2的表达来实现的。本研究为氧化苦参碱的抗肿瘤机制增添了新的内容。Objective To investigate the anticancer effect of oxymatrine on cervical cancer cell line （HeLa）. Mothocls MTT assay was used to detect the anti-proliferative effect of oxymatrine. Transwell chamber was used to detect the anti-metastatic effect of oxymatrine. Real-time PCR was used to detect the mRNA levels of MMP-2 and MMP-9. Western blot was used to detect the protein levels of MMP-2, MMP-9, AKT, p-AKT and GADPH. Plesults We found that application of oxymatrine significantly inhibited the growth of HeLa cells at the concentration above 0.8 mg/mL. We also found that oxymatrine （0.1, 0.2 and 0.4 mg/mL） inhibited the invasion of HeLa ceils under cytotoxic dose, which was （77.07± 20.43） %, （53.95 ± 18.17） % and （20.35±11.20） % of cells that migrated through the matrigel when compared with those of non-oxymatrine treatment group （P〈0.05）. Further research found that oxymatrine （0.1, 0.2 and 0.4 mg/mL） could reduce the expression of MMP-2 at the mRNA level, i.e. （82.76 -＋- 8.71）%, （39.51 ~ 12.79）% and （21.53±5. Z8）% of the expression level when compared with that of non-oxymatrine treatment group （P〈0.05）. The protein expression level of MMP-2 in 0.4 mg/mL group was （64.69 ± 16.52）% of non-oxymatrine treatment group （P〈0.05）. The phosphorylation level of AKT in 0.4 mg/mL group was （41.27±7. 13）% of non-oxymatrine treatment group （P〈0.05）. Oonclusion Oxymatrine can inhibit the invasion of HeLa cells by reducing the expression of MMP-2 via inhibiting the activity of AKT signal pathway. All together, our findings bring new insights into the mechanism of the anticancer effects induced by oxymatrine treatment.

关 键 词：氧化苦参碱 宫颈癌 侵袭 基质金属蛋白酶 

分 类 号：R737.33[医药卫生—肿瘤]