摘要
目的:蒺藜皂苷是我国的一种传统中药,本实验拟探讨蒺藜皂苷对非小细胞肺癌A549细胞增殖和侵袭能力的影响。方法:将A549细胞分为正常对照组和实验组(蒺藜皂苷250μg/mL组,蒺藜皂苷200μg/mL组,蒺藜皂苷150μg/mL组,蒺藜皂苷100μg/mL组,蒺藜皂苷50μg/mL组),采用CCK8法检测各组细胞的增殖活力,Transwell小室模型检测肿瘤细胞的侵袭能力,RT-PCR检测各组细胞中MMP9、caspase-3的mRNA表达差异。利用细胞免疫荧光染色观察各组细胞caspase-3荧光强度。结果:与正常对照组相比,蒺藜皂苷明显抑制A549细胞的增殖活性及侵袭能力,差异具有统计学意义(P<0.01)。在侵袭实验中,RT-PCR实验结果表明与对照组相比,蒺藜皂苷组MMP9表达明显降低,caspase-3表达明显增加,且都呈浓度依赖性,差异有统计学意义(P<0.01)。通过细胞免疫荧光染色实验发现,相比正常对照组,实验组caspase-3的荧光表达增强,其中高浓度皂苷组明显高于低浓度组。结论:蒺藜皂苷可以通过下调MMP9的表达以抑制A549细胞的侵袭能力,通过上调caspase-3表达来诱导细胞发生不可逆的凋亡。
Objective:To investigate the effects of Tribulus terrestris saponin on the proliferation and invasion ability of non⁃small cell lung cancer A549 cells.Methods:A549 cells were divided into normal control and experimental groups(Tribulus terrestris saponin 250μg/mL group,Tribulus terrestris saponin 200μg/mL group,Tribulus terrestris saponin 150μg/mL group,Tribulus terrestris saponin 100μg/mL group,Tribulus terrestris saponin 50μg/mL group).The proliferation viability of the cells in each group was detected by CCK8,the invasion of tumor cells was detected by Transwell analysis.The mRNA expression of MMP9 and caspase⁃3 in each group of cells was detected by RT⁃PCR.Immunofluorescence staining was used to observe the fluorescence intensity of caspase⁃3 in each group of cells.Results:Compared with the normal control group,Tribulus terrestris sapo⁃nin significantly inhibited the proliferation activity and invasion ability of A549 cells,which was statistically significant(P<0.01).In the invasion assay,compared with the control group,MMP9 expression was significantly reduced and caspase⁃3 expression was significantly increased in the Tribulus terrestris saponin group,in a concentration⁃dependent manner,with statistically significant differences(P<0.01).By cellular immunofluorescence staining experiments,it was found that the fluorescence expression of cas⁃pase⁃3 was enhanced in the experimental group compared with the normal control group,in which the high concentration saponin group was significantly higher than the low concentration group.Conclusion:Tribulus terrestris saponin can inhibit the invasive ability of A549 cells by downregulating the expression of MMP9,and induce irreversible apoptosis by upregulating the activation of caspase⁃3 expression.
作者
唐欢欢
黄亚楠
卢在庆
周向东
张海滨
李琪
刘合露
TANG Huan-huan;HUANG Ya-nan;LU Zai-qing;ZHOU Xiang-dong;ZHANG Hai-bin;LI Qi;LIU He-lu(Department of Respiratory Medicine,The First Affiliated Hospital of Hainan Medical College,Haikou 570102,China;Hainan Respiratory Disease Clinical Medical Center,Haikou 571199,China;Institute of Deep-sea Science and Engineering,Chinese Academy of Sciences,Sanya 572000,China;Key Laboratory of Mariculture,Ministry of Education,Ocean University of China,Qingdao 266000,China)
出处
《海南医学院学报》
2023年第23期1761-1766,共6页
Journal of Hainan Medical University
基金
国家重点研发计划(2022YFC2305004)
海南省重大科技专项(ZDKJ2021036)
海南省重点研发项目(ZDYF2020223)
海南省重点研发计划国际科技合作项目(GHYF2022011)
海南省创新团队项目(820CXTD448)
国家自然科学基金资助项目(82260001,82160012)。
关键词
蒺藜皂苷
非小细胞癌
含半胱氨酸的天冬氨酸蛋白水解酶
基质金属蛋白酶
Tribulus terrestris saponin
Non⁃small cell carcinoma
Cysteinyl aspartate specific proteinase
Matrix metallo⁃proteinase
