摘要
目的以肿瘤干细胞为切入点,基于Notch信号通路探究夏枯草对大肠癌细胞增殖及干性的调控机制。方法体外培养大肠癌SW480细胞,待细胞处于对数生长期后,随机分成对照组和0.1 mg/mL组、0.2 mg/mL组、0.4 mg/mL组。对照组不进行药物干预;剩余3组分别添加0.1 mg/mL、0.2 mg/mL、0.4 mg/mL的夏枯草药液,并将各组所含DMSO补足至等量,干预24 h后进行后续实验。采用倒置显微镜观察大肠癌SW480细胞生长形态及密度,0.4%台盼蓝染色法检测活细胞数并计算细胞存活率;CCK8法检测大肠癌SW480细胞的增殖抑制率;流式细胞术检测大肠癌SW480细胞群中干细胞比例;克隆球形成实验检测大肠癌SW480细胞的自我更新能力;Western blot法检测大肠癌SW480细胞内增殖调控蛋白c-Myc,肿瘤干细胞特征标记物CD44、CD133、SOX2以及Notch信号通路相关蛋白Notch1、Hes1蛋白相对表达量。结果与对照组比较,0.1 mg/mL组、0.2 mg/mL组、0.4 mg/mL组大肠癌SW480细胞生长和细胞存活率随着加药浓度增加而受到明显抑制(P<0.05);大肠癌SW480细胞增殖能力随着加药浓度增加而明显下降(P<0.05);大肠癌SW480细胞群中肿瘤干细胞比例随着加药浓度增加而明显下降(P<0.05);大肠癌SW480细胞自我更新能力随着加药浓度增加而明显抑制(P<0.05)。Western blot结果显示,与对照组相比,0.1 mg/mL组、0.2 mg/mL组、0.4 mg/mL组大肠癌SW480细胞增殖调控蛋白c-Myc,肿瘤干细胞特征性标记物CD44、CD133、SOX2蛋白及通路相关蛋白Notch1、Hes1的相对表达量随着加药浓度增加而明显下调(P<0.05)。结论夏枯草可以抑制大肠癌细胞的增殖及干性,抑制Notch通路的活化可能是其发挥作用的重要机制。
Objective From the perspective of colorectal cancer stem cells,the purpose of this study was to explore the effect and its mechanism of Spica Prunellae on the proliferation and stemness of colorectal cancer cells based on Notch signaling pathway.Methods Colorectal cancer SW480 cells were cultured in vitro.When the cells were in logarithmic growth phase,they were ran‐domly divided into control group,0.1 mg/mL group,0.2 mg/mL group and 0.4 mg/mL group.The control group did not receive drug intervention;the other three groups were added with 0.1 mg/mL,0.2 mg/mL and 0.4 mg/mL Spica Prunellae solution,respectively.Meanwhile,DMSO in each group was supplemented to the same amount.After 24 hours of intervention,the subsequent experiments were performed.The cell growth morphology and density were observed by inverted microscope.The number of viable cells was de‐tected by 0.4%Trypan blue staining and cell survival rate was calculated.The proliferation inhibition rate of colorectal cancer cells was detected using Cell Counting Kit-8(CCK8)assay.Flow cytometry was used to detect the proportion of stem cells in colorectal cancer cells.The self-renewal ability of colorectal cancer cells was detected by sphere formation assay.Western blot method was used to detect the relative expression of c-Myc,CD44,CD133,SOX2 and Notch pathway-related proteins Notch1 and Hes1 in colorectal cancer cells.Results Compared with the control group,the proliferation and viability of colorectal cancer SW480 cells in the 0.1,0.2,and 0.4 mg/mL groups were significantly inhibited with the increase of Spica Prunellae concentration(P<0.05).The proliferation ability of colorectal cancer SW480 cells significantly decreased with the increase of Spica Prunellae concentration(P<0.05).The proportion of cancer stem cells in colorectal cancer SW480 cells significantly decreased with the increase of Spica Prunel‐lae concentration(P<0.05).The self-renewal ability of colorectal cancer SW480 cells was significantly inhibited with the increase of Spica Prunellae
作者
田浩源
谢意
方翌
程瑛
林妍
苏代丰
魏丽慧
TIAN Haoyuan;XIE Yi;FANG Yi;CHENG Ying;LIN Yan;SU Daifeng;WEI Lihui(Innovation and Transformation Center,Fujian University of Traditional Chinese Medicine,Fuzhou,Fujian 350122,China;College of Pharmacy,Fujian University of Traditional Chinese Medicine,Fuzhou,Fujian 350122,China;Academy of Integrative Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou,Fujian 350122,China;Fujian Key Laboratory of Integrative Medicine on Geriatrics,Fuzhou,Fujian 350122,China)
出处
《康复学报》
CSCD
2024年第3期233-241,共9页
Rehabilitation Medicine
基金
国家自然科学基金青年项目(81703913,81603413)。
关键词
大肠癌
夏枯草
增殖
干性
NOTCH信号通路
colorectal cancer
Spica Prunellae
proliferation
stemness
Notch signaling pathway
