C1q样蛋白4对乳腺癌侵袭及转移影响的体内研究  

In Vivo Study of C1q-like Protein 4 on Invasion and Metastasis of Breast Cancer

作　　者：李潇[1] 赵博 郭研 徐壮语 徐繁[1] Li Xiao;Zhao Bo;Guo Yan(Department of Oncology,Affiliated Hospital of Chengde Medical University,Hebei Province,Chengde 067000,China)

机构地区：[1]河北省承德医学院附属医院肿瘤科,承德067000

出　　处：《华中科技大学学报(医学版)》2025年第2期203-210,共8页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：河北省2024年度医学科学研究课题(No.20240363)。

摘　　要：目的通过体内实验验证C1q样蛋白4(C1ql4)对NOD/SCID鼠乳腺癌转移瘤组织上皮间质转化(EMT)和干性特征的影响。方法构建免疫荧光素标记的稳转低表达和过表达C1ql4的细胞系(MDA-MB-231、MCF-7),尾静脉注射到4周龄雌性NOD/SCID鼠体内,IVIS活体成像检测转移瘤形成情况。注射8周后安乐死小鼠,取肺组织固定后石蜡包埋,苏木精-伊红(HE)染色观察转移灶数量;免疫组织化学染色(IHC)方法检测肺组织标本CD44、SOX2蛋白及ER、Her-2蛋白的表达水平;提取新鲜肺转移瘤中的总RNA和蛋白,qRT-PCR和Western blot法检测EMT和干细胞特性相关蛋白表达。结果IVIS活体成像结果显示:与MDA-MB-231组相比,MDA-MB-231-sh组小鼠体内肿瘤的荧光值及荧光密度显著降低(P<0.05);与MCF-7组相比,MCF-7-OE组小鼠体内肿瘤的荧光值及荧光密度显著升高(P<0.05)。小鼠肺内转移瘤组织HE染色及IHC结果显示:与MDA-MB-231组相比,MDA-MB-231-sh组小鼠肺内转移灶数量明显减少,且CD44、SOX2表达下调(均P<0.05),而ER及Her-2表达无明显变化;与MCF-7组相比,MCF-7-OE组小鼠肺内转移灶数量明显增多且CD44、SOX2表达增加(均P<0.05),ER及Her-2表达无明显变化。小鼠肺内转移瘤组织qRT-PCR和Western blot结果显示,MDA-MB-231-sh组干性相关蛋白和间质标志物表达量明显低于MDA-MB-231组(均P<0.05),但上皮标志物表达量则高于MDA-MB-231组(P<0.05);MCF-7-OE组干性相关蛋白和间质标志物表达量明显高于MCF-7组(均P<0.05),但上皮标志物表达量则低于MCF-7组(P<0.05)。结论C1ql4通过增强癌细胞干性表达和EMT进程促进小鼠乳腺癌肺转移瘤的形成和进展。Objective In vivo validation of the effects of C1q-like protein 4(C1ql4)on epithelial-mesenchymal transition(EMT)and stemness characteristics of breast cancer metastatic tissues in NOD/SCID mice.Methods Immunofluorescence-labeled cell lines with stable low-expression and over-expression of C1ql4(MDA-MB-231,MCF-7)were constructed,and then injected intravenously into 4-week-old female NOD/SCID mice.The formation of metastatic tumors was detected by IVIS live imaging.Eight weeks after injection,the mice were euthanized,and lung tissues were fixed in paraffin.HE staining was performed to observe the number of metastatic foci,and immunohistochemistry(IHC)was used to detect the expression levels of CD4,SOX2,ER,Her-2 protein in tissue specimens.Total RNA and protein were extracted from fresh lung metastatic tumors,and the expression of EMT-related proteins and stem cell-related proteins were detect by qRT-PCR and Western blotting.Results The IVIS live imaging results showed that,compared with MDA-MB-231 group,the fluorescence intensity and density of tumors in the MDA-MB-231-sh group were significantly decreased(P<0.05);compared with the MCF-7 group,the fluorescence intensity and density of tumors in the MCF-7-OE group were significantly increased(P<0.05).HE staining and IHC results of mouse lung metastatic tissues showed that,compared with the MDA-MB-231 group,the number of lung metastatic foci in the MDA-MB-231-sh group was significantly reduced,and the expressions of CD44 and SOX2 were downregulated(all P<0.05),with no significant changes in ER and Her-2 expression;whereas compared with the MCF-7 group,the number of lung metastatic foci in the MCF-7-OE group was significantly increased,and expressions of CD44 and SOX2 were upregulated(all P<0.05),with no significant changes in ER and Her-2 expression.The expression levels of stemness-related proteins and interstitial markers of mouse lung metastatic tissues detected by qRT-PCR and Western blotting in the MDA-MB-231-sh group were significantly lower than those in the

关 键 词：乳腺癌 C1q样蛋白4 上皮间质转化 干细胞特性 

分 类 号：R737.9[医药卫生—肿瘤]