摘要
目的观察重楼皂苷Ⅰ(polyphyllinⅠ, PPⅠ)对A549细胞转移的影响,并探讨其抑制细胞转移的分子机制。方法以人肺腺癌A549细胞为研究对象,利用台盼蓝活力检测实验和Transwell侵袭实验分别探究PPⅠ对肿瘤细胞增殖和转移的影响;采用Real-time quantitative PCR实验和Western Blot实验探究PPⅠ对基质金属蛋白酶-2(matrix metalloproteinase-2, MMP-2)、基质金属蛋白酶-9(matrix metalloproteinase-9, MMP-9)和核因子-κB(nuclear factor kappa B, NF-κB)的亚基p65 mRNA和蛋白的表达,并探讨了NF-κB/p65与MMP-2、MMP-9的关系。结果与空白对照组相比,PPⅠ给药组从0.4μmol·L^(-1)起,能够抑制A549细胞的增殖(P<0.05);PPⅠ给药组(0.8μmol·L^(-1))能够抑制A549细胞的侵袭(P<0.05);PPⅠ以浓度依赖性抑制MMP-2、MMP-9和p65 mRNA和蛋白的表达(P<0.05);且过表达p65后逆转了PPⅠ对MMP-2、MMP-9 mRNA的下调(P<0.05)。结论 PPⅠ可通过抑制NF-κB/p65通路下调下游MMP-2和MMP-9的表达,进而抑制A549细胞的转移。
Objective To observe the effects of PPⅠ on A549 cells metastasis and discuss the molecular mechanism of metastasis inhibited by PPⅠ. Methods The human lung adenocarcinoma A549 cells were taken as this research object. The inhibition of PPⅠin A549 proliferation and metastasis was explored by Trypan blue viability assay and transwell assay respectively. The effect of PPⅠon the mRNA and protein expression of matrix metalloproteinase-2(MMP-2), matrix metalloproteinase-9(MMP-9) and nuclear factor kappa B/p65(NF-κB/p65) were explored by Real-time quantitative PCR assay and western blot assay respectively. And the relationship between NF-κB/p65 and MMP-2, MMP-9 was also discussed. Results As compared with that of the blank control group, the proliferation of A549 was inhibited in a concentration dependent manner in the PPⅠgroups from 0.4 μmol·L^(-1)(P<0.05). The invasion of A549 cells was inhibited in the PPⅠgroup(0.8 μmol·L^(-1))(P<0.05). The mRNA and protein expressions of MMP-2, MMP-9 and p65 were reduced in the PPⅠgroups in a concentration dependent manner(P<0.05). Simultaneously, over-expression of p65 reversed the reduction of MMP-2 and MMP-9 mRNA expression caused by PPⅠ(P<0.05). Conclusion PPⅠcan down-regulateexpressions of MMP-2 and MMP-9 by inhibiting the expression of NF-κB/p65 so as to inhibit the metastasis of A549 cells.
作者
李龙妹
李秋萍
龙顺钦
河文峰
吴万垠
LI Longmei;LI Qiuping;LONG Shunqin;HE Wenfeng;WU Wanyin(The Second Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangdong Provincial Hospital of Traditional Chinese Medicine,Guangzhou 510370,Guangdong,China)
出处
《中华中医药学刊》
CAS
北大核心
2021年第8期149-152,I0031,共5页
Chinese Archives of Traditional Chinese Medicine
基金
国家自然科学基金青年科学基金(81803919)
广东省自然科学基金博士启动项目(2018A030310601)。
