摘要
目的:基于肿瘤微环境中转移相关因子探索健脾消癌方抑制结肠癌HCT116细胞增殖、侵袭及迁移的作用机制。方法:14只SPF级SD大鼠随机分为无药血清组及健脾消癌方组(10.8 g·kg^(-1)),每组7只,适应性饲养后,灌胃给予相应生理盐水和中药7 d,获取无药血清与健脾消癌方含药血清。HCT116细胞分为空白组、5%无药血清组、10%无药血清组、20%无药血清组、5%健脾消癌方含药血清组、10%健脾消癌方含药血清组、20%健脾消癌方含药血清组,分别处理24 h、48 h、72 h后采用CCK-8实验检测各组细胞的增殖能力,筛选最佳含药血清浓度和干预时间;Transwell实验检测细胞的侵袭及迁移能力;Western Blot及RT-PCR分别检测趋化因子受体4(chemokine receptor 4,CXCR4)、转化生长因子-β(transforming growth factor-β,TGF-β)、整合素αvβ5(integrinαvβ5,ITGαvβ5)、血清钙结合蛋白A4(calcium-binding protein A4,S100A4)、血清钙结合蛋白A8(calcium-binding protein A8,S100A8)、血清钙结合蛋白A9(calcium-binding protein A9,S100A9)等肿瘤微环境中转移相关因子的蛋白及mRNA表达水平。结果:不同浓度健脾消癌方含药血清均能抑制HCT116细胞增殖,以20%健脾消癌方含药血清干预细胞48 h的抑制作用最为明显(P<0.001)。与20%无药血清组比较,20%健脾消癌方含药血清能显著抑制HCT116细胞侵袭及迁移,显著降低CXCR4、TGF-β、S100A4、S100A8、S100A9、ITGαvβ5的蛋白与mRNA表达水平(P<0.05)。结论:健脾消癌方能抑制结肠癌HCT116细胞增殖、侵袭及迁移,其机制可能与降低CXCR4、TGF-β、S100A4、S100A8、S100A9、ITGαvβ5等肿瘤微环境中转移相关因子的表达有关。
Objective:Based on the metastasis related factors in the tumor microenvironment,to explore the mechanism of Jianpi Xiao'ai Formula in inhibiting the proliferation,invasion,and migration of colon cancer HCT116 cells.Method:14 SPF grade SD rats were randomly divided into a drug-free serum group and a Jianpi Xiaocancer Formula group(10.8 g·kg^(-1)),with 7 rats in each group.After adapting to feeding,they were given corresponding physiological saline and traditional Chinese medicine by gavage for 7 days to obtain drug-free serum and Jianpi Xiaocancer Formula containing serum.HCT116 cells were divided into a blank group,a 5%drug-free serum group,a 10%drug-free serum group,a 20%drug-free serum group,a 5%Jianpi Xiao'ao Formula medicated serum group,a 10%Jianpi Xiao'ao Formula medicated serum group,and a 20%Jianpi Xiao'ao Formula medicated serum group.After treatment for 24 hours,48 hours,and 72 hours,the proliferation ability of each group of cells was tested using the CCK8 experiment to screen the best intervention serum containing drugs and intervention time;Transwell experiment on the invasion and migration ability of cells;Western Blot and RT-PCR were used to detect chemokine receptor 4(CXCR4)and transforming growth factor,respectively-β(TGF-β),integrinαvβ5(ITGαvβ5).The protein and mRNA expression levels of metastasis related factors in tumor microenvironment include serum calcium binding protein A4(S100A4),serum calcium binding protein A8(S100A8),and serum calcium binding protein A9(S100A9).Results:Serum containing different concentrations of Jianpi Xiao'ao Formula can inhibit the proliferation of HCT116 cells,with 20%Jianpi Xiao'ao Formula containing serum intervening for 48 hours having the most significant inhibitory effect(P<0.05).Compared with the drug-free serum group,Jianpi Xiao'ai Formula can significantly inhibit the invasion and migration of HCT116 cells,and significantly reduce CXCR4 and TGF-β,S100A4,S100A8,S100A9,ITGavβ5 protein and mRNA expression levels(P<0.05).Conclusion:Jian Pi Xiao'ai Formula can inhibit the proliferation,invasion,and migration of colon cancer HCT116 cells,and its mechanism may be by reducing CXCR4 and TGF-β,S100A4,S100A8,S100A9,ITGavβ.The expression of metastasis related factors in grade 5 tumor microenvironment is achieved.
作者
何威华
邓兰
蒋益兰
HE Weihua;DENG Lan;JIANG Yilan(Wuhan Hospital of Traditional Chinese Medicine,Wuhan Hubei China 430050;Hunan University of Chinese Medicine,Changsha Hunan China 410208;The Affiliated Hospital to Jiangxi University of Traditional Chinese Medicine,Nanchang Jiangxi China 330006;The Affiliated Hospital to Hunan Academy of Traditional Chinese Medicine,Changsha Hunan China 410006)
出处
《中医学报》
CAS
2023年第12期2639-2646,共8页
Acta Chinese Medicine
基金
国家自然科学基金项目(81774287)
武汉市卫生健康委科研项目(WZ21Q19)
湖南省研究生创新科研项目(CX2018B477)
湖北省中医药管理局中医药科研项目(ZY2023F054)。
关键词
结肠癌
健脾消癌方
肿瘤微环境
细胞增殖
细胞侵袭
细胞迁移
大鼠
colon cancer
Jianpi Xiao'ai Formula
tumor micro-environment
cell proliferation
cell invasion
cell migration
rat
