摘要
目的探讨巨噬细胞表达的泛素连接酶F框/WD40域蛋白(FBXW7)对卵巢癌小鼠肿瘤生长及卵巢癌微环境的影响。方法选择野生型(对照组)和FBXW7基因敲除(实验组)的雌性C57BL/6J小鼠为实验动物,均为6~8周龄。蛋白质印迹法和逆转录定量-聚合酶链反应(qRT-PCR)检测2组小鼠FBXW7基因表达水平。实验组和对照组小鼠皮下均接种5×10^(5)个小鼠卵巢癌细胞株ID8构建卵巢癌模型,每组6只。每隔4天用游标卡尺测量肿瘤长径和短径,计算肿瘤体积。流式细胞术检测并对比2组小鼠肿瘤组织中CD4^(+)T细胞和CD8^(+)T细胞比例。蛋白质印迹法和qRT-PCR检测2组小鼠M1和M2型巨噬细胞特异性分子蛋白及mRNA表达水平。免疫组化检测肿瘤组织中转化生长因子β1(TGF-β1)和α-平滑肌肌动蛋白(α-SMA)的分布及表达。结果与对照组相比,实验组小鼠巨噬细胞中FBXW7蛋白和mRNA表达水平降低,差异有统计学意义,t值分别为24.418和26.728,均P<0.001.建立卵巢癌小鼠模型16 d后,实验组小鼠肿瘤体积为(341.74±163.66)mm^(3),大于对照组的(85.76±57.46)mm^(3),差异有统计学意义,t=3.615,P=0.011;实验组小鼠肿瘤组织中CD4^(+)T细胞比例为(16.49±1.11)%,CD8^(+)T细胞比例为(38.64±1.65)%,均高于对照组的(9.87±0.74)%和(12.26±0.90)%,差异有统计学意义(t=8.634,P=0.001;t=24.282,P<0.001)。与对照组小鼠相比,实验组小鼠肿瘤坏死因子-α(TNF-α,t=8.094,P=0.001)、趋化因子配体9(CXCL9,t=11.168,P<0.001)和CXCL10(t=14.437,P<0.001)蛋白表达均降低,TNF-α(t=12.826,P<0.001)、CXCL9(t=5.654.P<0.001)和CXCL10(t=11.979,P<0.001)mRNA表达水平均降低;精氨酸酶1(Arg-1,t=20.776,P<0.001)、血管内皮生长因子(VEGF,t=18.580,P<0.001)和CC型趋化因子配体17(CCL17,t=10.962,P<0.001)蛋白表达均升高,Arg-1(t=10.958,P<0.001)、VEGF(t=13.554,P=0.001)和CCL17(t=35.358,P<0.001)mRNA表达水平均升高。与对照组相比,实验组诱导型一氧化氮合酶(iNOS)蛋白表达积分光�
Objective To investigate the effect of ubiquitin ligase F-box and WD repeat domain containing 7(FBXW7)expressed by macrophages on tumor growth and microenvironment of ovarian cancer in mice.Methods Wild-type(control group)and FBXW7 gene knockout(experimental group)female C57BL/6J mice at six-to-eight weeks old were selected as experimental animals.Western blot and quantitative reverse transcription-polymerase chain reaction(qRT-PCR)were used to detect the expression level of FBXW7 gene in 2 groups of mice.Both the experimental group and the control group were inoculated subcutaneously with 5×10^(5) mouse ovarian cancer cell lines ID8 to construct ovarian cancer models,with 6 mice in each group.The length and short diameter of the tumor were measured with a vernier caliper every 4 days to calculate the tumor volume.Flow cytometry was used to detect and compare the ratio of CD4^(+) T cells and CD8^(+) T cells in the tumor tissues of the two groups of mice.Western blotting and qRT-PCR were used to detect the expression levels of specific molecular proteins and mRNA of M1 and M2 macrophages in the two groups of mice.Immunohistochemistry was used to detect the distribution and expression of transforming growth factorβ1(TGF-β1)and α-smooth muscle actin(α-SMA)in tumor tissues.Results Compared with the control group,the expression level of FBXW7 gene in the experiental group was lower,the difference was statistically significant,t values were 24.418 and 26.728,both P<0.001.Sixteen days after the establishment of ovarian cancer mouse model,the tumor volume of the experimental group was(341.74±163.66)mm^(3),which was greater than that of the control group(85.76±57.46)mm^(3),the difference was statistically significant(t=3.615,P=0.011).The proportion of CD4^(+)T cells in the tumor volume of the experimental group was(16.49±1.11)%,and the proportion of CD8^(+)T cells was(38.64±1.65)%,both of which were higher than those in the control group(9.87±0.74)%and(12.26±0.90)%,the differences were statistically significant
作者
徐洋
王媛
曹蔚
车瑾
付红星
夏俊
秦婷婷
XU Yang;WANG Yuan;CAO Wei;CHE Jin;FU Hong-xing;XIA Jun;QIN Ting-ting(Wuhan Third Hospital(Tongren Hospital Affiliated to Wuhan University),Wuhan 430060,China)
出处
《中华肿瘤防治杂志》
CAS
北大核心
2021年第20期1518-1525,共8页
Chinese Journal of Cancer Prevention and Treatment
基金
武汉市卫生计生委科研项目(WX19D41)。
关键词
巨噬细胞
FBXW7
卵巢癌
肿瘤微环境
极化
macrophages
FBXW7
ovarian cancer
tumor microenvironment
polarization
