摘要
目的观察穿心莲内酯（Andrographolide，AD）对宫颈癌细胞迁移和侵袭的抑制情况，并探讨其分子机制。方法体外培养宫颈癌细胞系HeLa细胞，经100nmol/L佛波酯（phorbol-12myristate-13acetate，PMA）诱导24h后，加入不同浓度（0，1，5，10μmol／L）的AD继续孵育24h。噻唑蓝比色（MTF）法检测HeLa细胞的生长抑制情况；小室侵袭实验分析AD对PMA诱导HeLa细胞的迁移与侵袭情况。Westernblot检测基质金属蛋白酶-9（matrix metalloproteinase-9，MMP-9）的表达以及核因子κB（Nuclear Factorkappa B，NF-κB）p65亚基的转位。结果0—10μmoL／LAD对HeLa细胞的生长增殖无明显毒性作用。10μmoL／LAD也能使HeLa细胞迁移和侵袭率分别降低49％和52％。Westernblot结果显示AD能以剂量依赖性方式抑制PMA诱导的MMP-9表达，并能以剂量依赖性方式抑制NF—κBp65亚基核转位。结论AD抑制NF—κB介导的MMP-9表达，从而影响宫颈癌细胞的迁移和侵袭。
Objective To observe the inhibitory effect of andrographolide （AD） on the migration and invasion of human cervical cancer cells, and investigate the molecular mechanisms underlying the inhibitory effect of AD. Methods Human cervical cancer cell lines HeLa cells were cuhured in vitro. After induced by 100 nmol/L of phorbol-12 - myristate-13 - acetate（PMA） for 24 h, different concentration of AD was co-incubated for another 24 h. Cell viability was detected by methyl thiazolyl tetrazoliym （MTT） assay. Chamber invasion assay was used to observe the migration and invasion, respectively. Expression of matrix metalloproteinase-9 （MMP-9） and translocation of nuclear factor kappa B （NF-κB） p65 subunit were detected by Western blot. Results The concentration （0 - 10 μmol/L） of AD exhibited a non-eytotoxic effect on HeLa cells. 10μmol/L of AD decreased PMA-indueed migration by 49% and invasion by 52%. Western blot showed that AD suppressed PMA-induced MMP-9 protein expression in a dose-dependent manner. Treatment of AD could decrease the translation of p65 in HeLa cells. Conclusions AD inhibits MMP-9 expression mediated by NF-κB, which lead to the suppression of migration and invasion of cervical cancer cells.
出处
《中国医师杂志》
CAS
2013年第3期304-306,共3页
Journal of Chinese Physician
关键词
穿心莲内酯
药理学
NF-κB
宫颈肿瘤
病理学
肿瘤细胞
培养的
病理学
ANDROGRAPHOLIDE/pharmacology
NF-kappa B
Uterine cervical neoplasms/pathology
Tumor cells, cultured/pathology
