桃叶珊瑚苷对人结肠癌HT-29细胞恶性生物学行为及SCAP/SREBP-1通路的影响  

Aucubin regulates malignant biological behavior of human colon cancer HT-29 cells possibly via SCAP/SREBP-1 pathway

作　　者：冯淑炯 江巧丽 FENG Shu-jiong;JIANG Qiao-li(Department of Gastroenterology,Hangzhou geriatrics hospital,Hangzhou,Zhejiang province,310015,China;Department of Gastroenterology,Zhejiang Tongde hospital,Hangzhou,Zhejiang province,310012,China)

机构地区：[1]杭州市老年病医院消化内科,杭州310015 [2]浙江省立同德医院消化科,杭州310012

出　　处：《浙江中西医结合杂志》2021年第6期509-514,共6页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

摘　　要：目的探讨桃叶珊瑚苷对人结肠癌HT-29细胞恶性生物学行为及固醇调节元件结合蛋白(SREBP)裂解激活蛋白(SCAP)/固醇调节元件结合蛋白-1(SREBP-1)通路的影响。方法实验设结肠癌HT-29细胞组、5-氟尿嘧啶组(30μmol/L)、桃叶珊瑚苷低剂量组(50μmol/L)、桃叶珊瑚苷高剂量组(100μmol/L),各组每孔设6个平行样,培养72h后,采用四唑盐(MTT)法测定细胞增殖,流式细胞仪测定细胞凋亡水平,Transwell腔室系统测定细胞侵袭迁移能力,实时荧光定量PCR及蛋白免疫印迹法测定细胞SCAP、SREBP-1 mRNA与蛋白水平。结果与结肠癌HT-29细胞组比较,5-氟尿嘧啶组、桃叶珊瑚苷低、高剂量组人结肠癌HT-29细胞OD值[(0.32±0.03)、(0.65±0.04)、(0.47±0.05)比(0.95±0.03),P均<0.05]及存活率[(29.47±3.14)%、(55.98±2.48)%、(38.45±2.54)%比(94.25±5.63)%,P均<0.05]下降,穿膜数[(159.69±24.84)个、(422.59±32.96)个、(242.59±31.55)个比(659.85±29.96)个,P均<0.05]减少,SCAP、SREBP-1 mRNA[SCAP:(1.63±0.28)、(4.96±0.32)、(2.86±0.31)比(6.25±0.32),P均<0.05;SREBP-1:(1.95±0.42)、(4.73±0.50)、(2.93±0.51)比(5.90±0.52),P均<0.05]和蛋白[SCAP:(0.27±0.06)、(0.83±0.11)、(0.52±0.09)比(1.26±0.18),P均<0.05;SREBP-1:(0.24±0.04)、(0.87±0.12)、(0.56±0.08)比(1.22±0.17),P均<0.05]水平降低,凋亡率[(15.96±0.78)%、(8.99±0.85)%、(12.63±0.87)%比(4.32±0.63)%,P均<0.05]升高。与5-氟尿嘧啶组比较,桃叶珊瑚苷低、高剂量组人结肠癌HT-29细胞OD值[(0.65±0.04)、(0.47±0.05)比(0.32±0.03),P均<0.05]及存活率[(55.98±2.48)%、(38.45±2.54)%比(29.47±3.14)%,P均<0.05]较高,穿膜数[(422.59±32.96)个、(242.59±31.55)个比(159.69±24.84)个,P均<0.05]增多,SCAP、SREBP-1 mRNA[SCAP:(4.96±0.32)、(2.86±0.31)比(1.63±0.28),P均<0.05;SREBP-1:(4.73±0.50)、(2.93±0.51)比(1.95±0.42),P均<0.05]和蛋白[SCAP:(0.83±0.11)、(0.52±0.09)比(0.27±0.06),P均<0.05;SREBP-1:(0.87±0.12)、(0.56±0.08)比(0.24±0.04),P均<0.05]水平升高,凋亡率Objective To investigate the effects of aucubin on the malignant biological behavior of human colon cancer HT-29 cells and the underlying mechanism.Methods HT-29 cells were divided into four groups:control,5-fluorouracil(30μmol/L),low dose(50μmol/L)and high dose(100μmol/L)of aucubin groups.Each group has 6 parallel samples/wells.After cells were treated for 72 hours,cell viability was determined by tetrazolium salt(MTT)assay,percentage of apoptotic cells was measured by flow cytometry,cell invasion and migration were detected using transwell chamber system and the m RNA and protein levels of sterol regulatory element binding protein(SREBP)cleavage activating protein(SCAP)and SREBP-1 were assessed by real-time reverse transciption PCR and Western blotting.Results Compared with the control group,the cells in the 5-fluorouracil,low and high doses aucubin groups showed decreased MTT OD values[(0.32±0.03),(0.65±0.04),(0.47±0.05)vs(0.95±0.03),P<0.05),decreased survival rates[(29.47±3.14)%,(55.98±2.48)%,(38.45±2.54)%vs(94.25±5.63)%,P<0.05],decreased number of transmembrane cells[(159.69±24.84),(422.59±32.96),(242.59±31.55)vs(659.85±29.96),P<0.05],decreased mRNA levels of SCAP and SREBP-1[SCAP:(1.63±0.28),(4.96±0.32),(2.86±0.31)vs(6.25±0.32),P<0.05;SREBP-1:(1.95±0.42),(4.73±0.50),(2.93±0.51)vs(5.90±0.52),P<0.05],and decreased protein levels of SCAP and SREBP-1[SCAP:(0.27±0.06),(0.83±0.11),(0.52±0.09)vs(1.26±0.18),P<0.05;SREBP-1:(0.24±0.04),(0.87±0.12),(0.56±0.08)vs(1.22±0.17),P<0.05],but increased percentage of apoptotic cells[(15.96±0.78)%,(8.99±0.85)%,(12.63±0.87)%vs(4.32±0.63)%,P<0.05].However,the degrees of growth inhibition,suppression of cell migration,decreased SCAP and SREBP-1 expression and induction of apoptosis in 5-fluorouracil group were more remarkable than those in both low and high doses aucubin groups.Moreover,the effects of aucubin on HT-29 cells were in a dose-dependent manner.Conclusion Aucubin can significantly inhibit either cell proliferation or invasion and promo

关 键 词：桃叶珊瑚苷 结肠癌 HT-29 恶性生物学 SCAP/SREBP-1通路 

分 类 号：R285.5[医药卫生—中药学]