理冲汤干预卵巢癌肿瘤微环境的机制研究  

Mechanism of Lichong Decoction intervened tumor microenvironment of ovarian cancer

作　　者：王冬梅[1] 庞瑞[1] WANG Dongmei;PANG Rui(Department of Western and Traditional Chinese Medicine,the Affiliated Tumor Hospital,Xinjiang Medica University,Urumqi 830011,China)

机构地区：[1]新疆医科大学附属肿瘤医院中西医结合科,乌鲁木齐830011

出　　处：《新疆医科大学学报》2022年第1期104-110,共7页Journal of Xinjiang Medical University

基　　金：新疆维吾尔自治区自然科学基金(2018D01C248)。

摘　　要：目的研究理冲汤含药血清对小鼠卵巢上皮癌ID8细胞抑制增殖和促进凋亡的作用,并观察理冲汤含药血清对于脐静脉内皮细胞(human umbilical vein endothelial cell,HUVECs)血管成形过程的作用,为临床应用提供实验依据。方法采用3-(4,5-二甲基噻唑-2)-2,5二苯基四氮唑溴盐(MTT)法检测理冲汤含药血清对ID8细胞增殖的影响;采用流式细胞术检测理冲汤含药血清对ID8细胞凋亡作用的影响;将HUVEC分别与理冲汤含药血清、调节性T细胞(regulatory T cells,Tregs)及抗血管内皮生长因子(vascular endothelial growth factor,VEGF)抗体进行共培养,分为8组进行:A,空白组(完全培养基);B,抗VEGF组(完全培养基+VEGF抗体);C,理冲汤含药血清组(理冲汤含药血清培养基);D,抗VEGF+理冲汤含药血清组(VEGF抗体+理冲汤含药血清培养基);E,Tregs组(完全培养基+Tregs);F,Tregs+抗VEGF组(完全培养基+Tregs+VEGF抗体);G,Tregs+含药血清组(Tregs细胞+含药血清培养基);H,Tregs+抗VEGF+理冲汤含药血清组(Tregs+VEGF抗体+理冲汤含药血清培养基)。通过小管成形实验观察各组血管生成情况差异,免疫荧光检测各组VEGF、基质金属蛋白酶2(Matrix metalloproteinase2,MMP-2)和基质金属蛋白酶9(Matrix metalloproteinase9,MMP-9)的表达情况。结果与空白组比较,理冲汤含药血清能够抑制ID8细胞的增殖,并且可以促进ID8细胞凋亡(P<0.05)与未加入Tregs各组相比,加入Tregs各组HUVECs呈现出明显的血管网状结构,而理冲汤含药血清组血管成网则不明显,VEGF抗体联合含药血清组血管成网情况最差;与未加入Tregs各组相比,加入Tregs各组VEGF、MMP-2和MMP-9的表达明显增高,理冲汤含药血清组表达较低,VEGF抗体联合理冲汤含药血清组表达最低。结论理冲汤对小鼠卵巢上皮癌ID8细胞有明显的抑制作用,并且可以促进其凋亡。同时理冲汤还可通过抑制VEGF、MMP-2和MMP-9的表达来抗血管的形成。Objective To explore the efficacy of Lichong decoction on proliferation and apoptosis of mouse ovarian epithelial cells of ID-8,and to observe the effect of Lichong decoction on the angiogenesis of human umbilical vein endothelial cells(HUVECs),which can provide an experimental basis for clinical application.Methods Ovarian epithelial cells ID8 were tested by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide for proliferation;The effect of Lichong decoction on apoptosis of ID8 cells were detected with flow cytometry.Drug-containing serum cells of Lichong decoction,regulatory T cells(Tregs)and anti-vascular endothelial growth factor(VEGF)antibodies were respectively co-cultured with HUVEC cells,divided into the following 8 groups:A,Blank group(complete medium);B,Anti-vegf group(complete medium+VEGF antibody);C,Drug-containing serum group(drug-containing serum medium);D,Anti-vegf+drug-containing serum group(VEGF antibody+drug-containing serum medium);E,Tregs group(complete medium+Tregs cells);F,Tregs+anti-VEGF group(complete medium+Tregs cells+VEGF antibody);G,Tregs+drug-containing serum group(Tregs cells+drug-containing serum medium);H,Tregs+anti-VEGF+drug-containing serum group(Tregs cells+VEGF antibody+drug-containing serum medium).The difference of angiogenesis was observed by tube forming experiment in each groups,and the expression of VEGF,MMP-2 and MMP-9 was detected by immunofluorescence in the groups.Results The drug containing serum could inhibit the proliferation of ID8 cell line and promote the apoptosis of ID8 cell line(P<0.05).Compared with those without Tregs cells,HUVECs in Tregs treated groups were significantly higher than those without Tregs cells.Compared with the group without Tregs cells,the expression of VEGF,MMP-2 and MMP-9 in Tregs group were significantly increased,the expression of VEGF,MMP-2 and MMP-9 in the drug containing serum group were lower than that in the drug containing serum group,and the lowest in VEGF antibody combined with drug containing serum group.Conclusi

关 键 词：卵巢癌 肿瘤微环境 调节性T细胞 

分 类 号：R273[医药卫生—中西医结合]