A novel mouse model of testicular granulosa cell tumors
A novel mouse model of testicular granulosa cell tumors
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DOI：
10.1093/molehr/gay023
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发表时间：
2018-07-01
影响因子：
4
通讯作者：
Li, Qinglei
中科院分区：
文献类型：
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作者：
Fang, Xin;Ni, Nan;Li, Qinglei
研究问题：在睾丸性索间质肿瘤的发生中，转化生长因子β（TGFB）信号失调的作用是什么？总结回答：TGFB信号的过度激活导致类似于颗粒细胞瘤（GrCTs）的睾丸肿瘤的发生。已知：在早期的研究中，我们证明了卵巢体细胞中组成性激活的TGFB受体1（TGFBR 1）促进了卵巢GrCTs的发生。研究设计、规模、持续时间：为了确定TGFB信号转导失调对睾丸的影响，我们使用抗苗勒管激素受体2型（Amhr 2）-Cre重组酶产生了具有组成性激活TGFBR 1的小鼠。研究了组成型活性TGF β R 1对睾丸发育的影响和睾丸肿瘤形成的时间轴。我们进一步研究了睾丸肿瘤的分子特征，并确定了已知参与睾丸GrCT发育的β-连环蛋白（CTNNB 1）的表达。e.从1周到6个月）与对照组一起沿着。收集睾丸样本并进行组织学和分子分析，包括苏木精和伊红（H和E）染色，实时PCR，免疫组织化学，免疫荧光和蛋白质印迹。使用每个基因型至少三只动物进行免疫染色/免疫印迹和实时PCR实验。数据表示为平均值+/- SEM。采用非配对双尾t检验确定统计学显著性，并在P值<0.05时报告。主要结果和机会作用：睾丸中携带组成型活性TGFBR 1的小鼠发生类似睾丸GrCT的肿瘤，这是睾丸中的一种罕见类型的肿瘤。睾丸肿瘤的形成导致细胞增殖改变、生殖细胞丢失和精子发生缺陷。免疫组化结果显示，这些肿瘤呈阳性表达，包括INHA、FOXO 1，更重要的是FOXL 2，FOXL 2是一种在卵巢中特异性表达的蛋白质，是正常颗粒细胞分化和功能所必需的。与免疫组化结果一致，FOXL 2蛋白仅在TGFBR 1-CA（Acre）小鼠的睾丸中检测到，但通过蛋白质印迹法在对照组中检测不到，表明支持细胞命运的潜在改变。为了探讨TGFBR 1过度激活的促肿瘤作用的潜在机制，我们检测了CTNNB 1的表达。结果显示，TGFBR 1-CA（Acre）小鼠睾丸肿瘤中CTNNB 1的表达增加。总的来说，这项研究揭示了增强的TGFB信号在tests.LARGE-SCALE数据的致瘤功能：N/A的局限性，原因回避：这项研究是使用小鼠进行的，实验范式和人类睾丸GrCTs的发现的直接相关性有待进一步研究。值得注意的是，组成型激活TGFBR 1被用来增强TGFB/SMAD信号传导活性，可能不会被解释为遗传原因的diseases.WIDER的影响的发现：这种小鼠模型可能被证明是一个有用的除了小鼠遗传学工具包的GrCT研究。我们发现TGFB信号失调导致睾丸GrCT的发育，这支持细胞和颗粒细胞之间的共同起源，并强调了平衡TGFB信号在生殖和发育中的至关重要性。研究资助/竞争兴趣（S）：这项研究得到了尤尼斯·肯尼迪·施莱佛国家儿童健康研究所的国家卫生研究所R 03 HD 082416基金的支持。人类发展和新的教师启动基金从得克萨斯A&M大学授予Q. L。作者声明没有竞争利益。
STUDY QUESTION: What is the role of dysregulated transforming growth factor beta (TGFB) signaling in the development of sex cord-stromal tumors in the testis?SUMMARY ANSWER: Overactivation of TGFB signaling results in the development of testicular tumors resembling granulosa cell tumors (GrCTs).WHAT IS KNOWN ALREADY: In an earlier study, we demonstrated that constitutively active TGFB receptor 1 (TGFBR1) in ovarian somatic cells promotes the development of ovarian GrCTs. However, the consequence of dysregulation of TGFB signaling in the pathobiology of the testis, remains poorly defined.STUDY DESIGN, SIZE, DURATION: To identify the impact of dysregulation of TGFB signaling on the testis, we generated mice with constitutive activation of TGFBR1 using anti-Mullerian hormone receptor type 2 (Amhr2)-Cre recombinase. The effect of constitutively active TGFBR1 on testis development and the timeline of testicular tumor formation were examined. We further investigated the molecular features of testicular tumors and determined the expression of beta-catenin (CTNNB1) known to be involved in testicular GrCT development.PARTICIPANTS/MATERIALS, SETTING, METHODS: Male mice with constitutive activation of TGFBR1 were examined at various developmental stages (i. e. from 1 week up to 6 months) along with controls. Testis samples were collected and processed for histological and molecular analyses, including haematoxylin and eosin (H and E) staining, real-time PCR, immunohistochemistry, immunofluorescence and western blotting. Immunostaining/immunoblotting and real-time PCR experiments were performed using at least three animals per genotype. Data are presented as mean +/- SEM. Statistical significance was determined using unpaired two-tail t-test and reported when P value is < 0.05.MAIN RESULTS AND THE ROLE OF CHANCE: Mice harboring constitutively active TGFBR1 in the testes developed tumors resembling testicular GrCTs, a rare type of tumors in the testis. The formation of testicular tumors led to altered cell proliferation, loss of germ cells and defective spermatogenesis. Immunohistochemically, these tumors were positive for inhibin alpha (INHA), forkhead box O1 (FOXO1), and more importantly, forkhead box L2 (FOXL2), a protein specifically expressed in the ovary and required for normal granulosa cell differentiation and function. Consistent with the immunohistochemical findings, FOXL2 proteins were only detectable in testes of TGFBR1-CA(Acre) mice but not those of controls by western blotting, suggesting potential alteration of Sertoli cell fate. To explore mechanisms underlying the tumor-promoting effect of TGFBR1 overactivation, we examined the expression of CTNNB1. The results revealed increased expression of CTNNB1 in testicular tumors in TGFBR1-CA(Acre) mice. Collectively, this study uncovered tumorigenic function of enhanced TGFB signaling in the testis.LARGE-SCALE DATA: N/A.LIMITATIONS, REASONS FOR CAUTION: This study was performed using mice, and the direct relevance of the experimental paradigm and findings to human testicular GrCTs awaits further investigation. Of note, constitutive activation of TGFBR1 was employed to enhance TGFB/SMAD signaling activity and may not be interpreted as the genetic cause of the disease.WIDER IMPLICATIONS OF THE FINDINGS: This mouse model may prove to be a useful addition to the mouse genetics toolkit for GrCT research. Our finding that dysregulation of TGFB signaling results in the development of testicular GrCTs supports a common origin between Sertoli cells and granulosa cells, and highlights the paramount importance of balanced TGFB signaling in reproduction and development.STUDY FUNDING/COMPETING INTEREST(S): This research was supported by the National Institutes of Health grant R03HD082416 from the Eunice Kennedy Shriver National Institute of Child Health & Human Development and the New Faculty Start- up Funds from Texas A&M University awarded to Q.L. The authors declare no competing interest.