Hopx报告基因小鼠谱系示踪乳腺干细胞  

Lineage tracing for mammary stem cells using Hopx reporter mice

作　　者：石建云 李文静 彭映 贾振华 张书锦 谭露露 原一桐 杜若琛 Shi Jianyun;Li Wenjing;Peng Ying;Jia Zhenhua;Zhang Shujin;Tan Lulu;Yuan Yitong;Du Ruochen(Key Laboratory of Cellular Physiology at Shanxi Medical University,Ministry of Education,Endocrine and Metabolic Diseases Key Laboratory of Shanxi Province,and Department of Physiology,Shanxi Medical University,Taiyuan 030001,Shanxi Province,China;Key Laboratory of Cellular Physiology at Shanxi Medical University,Ministry of Education,and Laboratory Animal Center,Shanxi Medical University,Taiyuan 030001,Shanxi Province,China)

机构地区：[1]山西医科大学细胞生理学教育部重点实验室,内分泌代谢病山西省重点实验室,山西医科大学生理学系,山西省太原市030001 [2]山西医科大学细胞生理学教育部重点实验室,山西医科大学动物中心,山西省太原市030001

出　　处：《中国组织工程研究》2025年第13期2720-2727,共8页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(82002063),项目负责人:石建云;山西省青年科学基金项目(201901D211316,201901D211319,20210302124172),项目负责人:石建云,原一桐,杜若琛;山西省级博士启动基金项目(SD1809),项目负责人:石建云;山西医科大学校级博士启动基金项目(XD1809),项目负责人:石建云;内分泌代谢病山西省重点实验室开放基金项目(202104010910009),项目负责人:石建云。

摘　　要：背景:乳腺干细胞对于乳腺组织的发育和稳态维持十分重要。乳腺癌的发生与乳腺干细胞有着紧密联系。最新研究表明,Hopx作为形态发生和细胞分化的重要转录调节因子已被证实在神经、肠道、毛囊等多种成体干细胞中表达,然而其在乳腺中的作用至今尚未见报道。目的:探究Hopx是否可以作为乳腺干细胞的特异性标志物。方法:①选取8周龄雌性Hopx-LacZ转基因小鼠,利用β-半乳糖苷酶染色检测Hopx在乳腺组织中的本底表达情况。②选取4周龄、6周龄、8周龄及妊娠14.5 d的雌性野生型小鼠,分别进行乳腺全组织洋红染色及K14、K8免疫荧光染色。③选取8周龄与妊娠17.5 d的雌性Hopx-CreERT2;Rosa26LacZ转基因小鼠,进行乳腺β-半乳糖苷酶染色。④选取4周龄雌性Hopx-CreERT2;Rosa26LacZ转基因小鼠,通过腹腔注射他莫昔芬(隔天注射1次,注射3次)来激活Cre/loxp系统,注射后4周进行乳腺β-半乳糖苷酶染色;选取8周龄雌性Hopx-CreERT2;Rosa26LacZ转基因小鼠,通过腹腔注射他莫昔芬(隔天注射1次,注射3次)来激活Cre/loxp系统,末次注射后4,10周进行乳腺β-半乳糖苷酶染色。⑤选取8周龄雌性Hopx-CreERT2;Rosa26LacZ转基因小鼠,通过腹腔注射他莫昔芬(隔天注射1次,注射3次)来激活Cre/loxp系统,注射后2周使Hopx-CreERT2;Rosa26LacZ转基因小鼠怀孕,对第1次妊娠17.5 d、第3次妊娠17.5 d的小鼠乳腺组织进行β-半乳糖苷酶染色。结果与结论:①β-半乳糖苷酶染色结果显示,8周龄Hopx-LacZ转基因小鼠的乳腺导管内确实含有Hopx阳性细胞且位于基底上皮,数量较少。②乳腺全组织染色及免疫荧光染色结果显示,野生型小鼠的乳腺在青春期、成熟期及妊娠期等不同发育阶段具有各自相应时期依赖性的特征,并且经历了一系列复杂的上皮重塑过程。③β-半乳糖苷酶染色结果显示,与8周龄雌性Hopx-CreERT2;Rosa26LacZ转基因小鼠相比,妊娠17.5 d的Hopx-CBACKGROUND:Mammary stem cells are vital for the development and homeostasis of mammary gland tissue.The occurrence of breast cancer has a close relationship with the mammary stem cells.Recent studies have shown that Hopx,as an important transcriptional regulator of morphogenesis and cell differentiation,has been confirmed to be expressed in a variety of adult stem cells such as nerves,intestines,hair follicles and lungs.However,its role in mammary stem cells has not been reported so far.OBJECTIVE:To investigate whether Hopx expression marks mammary stem cells.METHODS:(1)Female Hopx-LacZ transgenic mice aged 8 weeks were selected to detect the background expression of Hopx in breast tissue byβ-galactosidase staining.(2)Female wild-type mice at 4,6,and 8 weeks of age and 14.5 days of gestation were selected for whole-tissue magenta staining and K14 and K8 immunofluorescence staining,respectively.(3)Female Hopx-CreERT2;Rosa26LacZ transgenic mice aged 8 weeks and 17.5 days of gestation were selected and stained with breastβ-galactosidase.(4)The 4-week-old female Hopx-CreERT2;Rosa26LacZ transgenic mice were selected.The Cre/loxp system was activated by intraperitoneal injection of tamoxifen(once every other day,three times),and breastβ-galactosidase staining was performed 4 weeks after injection.The 8-week-old female Hopx-CreERT2;Rosa26LacZ transgenic mice were selected.The Cre/loxp system was activated by intraperitoneal injection of tamoxifen(once every other day,three times),and breastβ-galactosidase staining was performed 4 and 10 weeks after the last injection.(5)Female Hopx-CreERT2;Rosa26LacZ transgenic mice aged 8 weeks were selected.The Cre/loxp system was activated by intraperitoneal injection of tamoxifen(once every other day,three times).Hopx-CreERT2;Rosa26LacZ transgenic mice were pregnant 2 weeks after injection.The mammary tissue of mice at 17.5 days of the first pregnancy and 17.5 days of the third pregnancy was stained withβ-galactosidase.RESULTS AND CONCLUSION:(1)The results ofβ-galactosidase st

关 键 词：Hopx 乳腺干细胞 谱系追踪 妊娠期 腺泡形成 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] Q26[生物学—细胞生物学]