摘要
目的 探究拟黑多刺蚁活性组分促进结直肠癌SW116细胞凋亡的作用机制。方法 采用CCK8法、划痕实验、凋亡实验检测拟黑多刺蚁活性组分对SW116细胞增殖、迁移和凋亡的影响,Western blot法检测拟黑多刺蚁活性组分对SW116细胞Bax、Bcl-2蛋白表达的影响,RT-qPCR法检测miR-186-5p表达,双荧光素酶报告基因实验验证miR-186-5p与GJA1的靶向关系;过表达/抑制miR-186-5p,并检测Cx43蛋白表达;过表达Cx43,并检测拟黑多刺蚁活性组分对凋亡及相关蛋白、PI3K/Akt信号通路的影响。结果 拟黑多刺蚁活性组分能抑制SW116细胞增殖和迁移能力、Bcl-2蛋白表达,促进细胞凋亡、Bax蛋白和miR-186-5p表达。GJA1是miR-186-5p的下游靶基因,过表达miR-186-5p可降低SW116细胞Cx43蛋白表达,抑制miR-186-5p表达可升高SW116细胞Cx43蛋白表达,抑制miR-186-5p表达可逆转拟黑多刺蚁活性组分对Cx43表达的抑制作用。过表达Cx43可逆转拟黑多刺蚁活性组分对SW116细胞凋亡及相关蛋白、PI3K/Akt信号通路的影响。结论 拟黑多刺蚁活性组分抑制SW116细胞恶性生物学行为,通过miR-186-5p/Cx43调控PI3K/Akt信号通路,促进结直肠癌SW116细胞凋亡。
AIM To explore the mechanism of active components of Polyrhachis vicina Roger on apoptosis activation in colorectal cancer SW116 cells.METHODS The active components of P.vicina had their effects on viability,proliferation,migration and apoptosis of SW116 cells detected by CCK8 assay,scratch assay and apoptosis assay;and their impacts on the expressions of Bax and Bcl-2 proteins in SW116 cells were determined by Western blot.With its expression detected by RT-qPCR,miR-186-5p had its targeting relationship with GJA1 verified by a double luciferase reporter gene experiment,and its effect of overexpression/inhibition on Cx43 protein expression assessed as well.And the effects of Cx43 overexpression on cell apoptosis,apoptosis related proteins and PI3 K/Akt signaling pathway of active components of P.vicina were investigated.RESULTS The active components of P.vicina inhibited the proliferation and migration abilities of SW116 cells,Bcl-2 protein expression,and promoted apoptosis and the expressions of Bax protein and miR-186-5p.Targeting at the downstream GJA1,miR-186-5p inhibited Cx43 protein expression in SW116 cells at overexpression level,and increased Cx43 protein expression in SW116 cells at inhibition level.Inhibited miR-186-5p expression offset the inhibitory effect on Cx43 protein induced by active components of P.vicina.Cx43 overexpression offset the effects of active P.vicina components on apoptosis,apoptosis related proteins and PI3 K/Akt signaling pathway of SW116 cells.CONCLUSION The active components of P.vicina can inhibit the malignant biological behavior of colorectal cancer SW116 cells and activate their apoptosis via regulating PI3 K/Akt signaling pathway through miR-186-5p/Cx43.
作者
韦贤
李冬梅
何俊慧
韦洁
李懿
韦冬梅
王金妮
韦桂宁
WEI Xian;LI Dong-mei;HE Jun-hui;WEI Jie;LI Yi;WEI Dong-mei;WANG Jin-ni;WEI Gui-ning(Youjiang Medical University For Nationalities,Baise 533000,China;Guangxi Institute of Chinese Medicine and Pharmaceutical Sciences,Nanning 530022,China)
出处
《中成药》
CAS
CSCD
北大核心
2022年第6期1783-1791,共9页
Chinese Traditional Patent Medicine
基金
广西自然科学基金项目(2018GXNSFDA281046)
广西基地与人才专项(桂科AD22035055)。
