白花蛇舌草提取物通过下调Hippo-YAP信号通路促进结肠癌细胞凋亡  

Extract of Baihuasheshecao(Hedyotis diffusa)Improving Apoptosis of Colon Cancer HCT-8 Cells by Down-regulating Hippo-YAP Signaling Pathway

作　　者：廖德贵[1] 曾嘉敏 赖妙玲[1] 郅程[1] 唐甜[1] LIAO Degui;ZENG Jiamin;LAI Miaoling;ZHI Cheng;TANG Tian(The Second Affiliated Hospital of Guangzhou Medical University,Guangzhou 510260,Guangdong,China)

机构地区：[1]广州医科大学附属第二医院,广东广州510260

出　　处：《中华中医药学刊》2022年第2期248-251,I0036-I0038,共7页Chinese Archives of Traditional Chinese Medicine

基　　金：广东省中医药局科研项目(20182085)。

摘　　要：目的探讨白花蛇舌草提取物熊果酸、齐墩果酸对结肠癌细胞增殖与凋亡的影响及其机制。方法不同浓度的熊果酸、齐墩果酸作用结肠癌(HCT-8)细胞24、48、72 h后,CCK-8法、平板克隆形成实验检测HCT-8细胞的存活情况。筛选最佳实验条件,将细胞分为对照组和实验组,各组细胞做成细胞爬片,细胞免疫荧光检测YAP1蛋白水平。各组细胞消化离心做成细胞蜡块,免疫组织化学方法检测YES相关蛋白(YAP1)、p53基因(p53)、尾型同源盒转录因子2(CDX-2)、白介素6(IL-6)、第10号染色体缺失的磷酸酶和张力蛋白同源等位基因(PTEN)、转化生长因子-β1(TGF-β1)、肿瘤坏死因子-α(TNF-α)的蛋白水平。结果白花蛇舌草提取物可不同程度抑制结肠癌HCT-8细胞的增殖。细胞免疫荧光结果显示,对照组YAP1蛋白表达较实验组强。免疫组化结果显示YAP1的表达与TNF-α的表达有一定相关性。结论白花蛇舌草提取物通过抑制Hippo-YAP信号通路的活性,抑制结肠癌HCT-8细胞的增殖,诱导其凋亡。白花蛇舌草提取物可能是一种潜在的肠癌治疗药物。Objective To investigate the effects of ursolic acid and oleanolic acid on the proliferation and apoptosis of colon cancer cells.Methods The survival of colon cancer HCT-8 cells was detected by CCK-8 and plate cloning after treatment with ursolic acid and oleanolic acid at different concentrations for 24,48 and 72 h.The optimal experimental conditions were selected and the cells were divided into control group and experimental group.Cells in each group were made into cell slides and the level of YAP1 protein was detected by cell immunofluorescence.Cells in each group were digested and centrifuged to make cell wax blocks.Immunohistochemistry method was used to detect the protein levels of YAP1,CDX-2,IL-6,PTEN,TGF-βand TNF-α.Results The extracts from Baihuasheshecao(Hedyotis diffusa)could inhibit the proliferation of HCT-8 cells in colon cancer.The results of cell immunofluorescence showed that the expression of YAP1 protein in the control group was stronger than that in the experimental group,and also showed that the expression of YAP1 was correlated with the expression of TNF-α.Conclusion The extract of Baihuasheshecao(Hedyotis diffusa)could inhibit the proliferation of colon cancer HCT-8 cells and induce their apoptosis by inhibiting the activity of Hippo-YAP signaling pathway.The extract of Baihuasheshecao(Hedyotis diffusa)may be a potential drug for the treatment of colon cancer.

关 键 词：白花蛇舌草提取物 Hippo信号通路 结肠癌细胞 凋亡 

分 类 号：R284.2[医药卫生—中药学]