Sox30基因敲除致小鼠睾丸间质细胞增殖异常的研究

唐莹; 姜晓; 韩飞; 周洋西; 崔志鸿; 刘晋祎; 杨惠芳; 曹佳

doi:10.16016/j.1000-5404.202102044

您当前的位置：

首页 >

文章列表页 >

Sox30基因敲除致小鼠睾丸间质细胞增殖异常的研究

基础医学 | 更新时间：2026-01-09

- Sox30基因敲除致小鼠睾丸间质细胞增殖异常的研究
- Abnormal proliferation of Leydig cells in Sox30 knockout mice
- 第三军医大学学报 2021年43卷第11期页码：1063-1073
- 作者机构：
  
  1.750004 银川，宁夏医科大学公共卫生与管理学院劳动卫生与环境卫生教研室
  2.400038 重庆，陆军军医大学(第三军医大学)军事预防医学系毒理学研究所
  3.400700 重庆，西南大学药学院中医药学院
- 作者简介：
  
  [杨惠芳，E-mail：joyceyhf@163.com YANG Huifang, E-mail: joyceyhf@163.com
  [曹佳，E-mail：caojia1962@126.com CAO Jia, E-mail: caojia1962@126.com
- 基金信息：
  
  国家自然科学基金重点项目(81630087);重庆市基础科学与前沿技术研究重点项目(cstc2017jcyjBX0064)
- DOI：10.16016/j.1000-5404.202102044
  中图分类号： R322.64;R329.28;R394.3
- 收稿：2021-02-06，
  
  修回：2021-03-26，
  
  纸质出版：2021-06-15
- 稿件说明：
移动端阅览
唐莹, 姜晓, 韩飞, 等. Sox30基因敲除致小鼠睾丸间质细胞增殖异常的研究[J]. 第三军医大学学报, 2021,43(11):1063-1073.

Ying TANG, Xiao JIANG, Fei HAN, et al. Abnormal proliferation of Leydig cells in Sox30 knockout mice[J]. Journal of Third Military Medical University, 2021, 43(11): 1063-1073.
唐莹, 姜晓, 韩飞, 等. Sox30基因敲除致小鼠睾丸间质细胞增殖异常的研究[J]. 第三军医大学学报, 2021,43(11):1063-1073. DOI： 10.16016/j.1000-5404.202102044.

Ying TANG, Xiao JIANG, Fei HAN, et al. Abnormal proliferation of Leydig cells in Sox30 knockout mice[J]. Journal of Third Military Medical University, 2021, 43(11): 1063-1073. DOI： 10.16016/j.1000-5404.202102044.

摘要

目的

探讨Sox30基因敲除对小鼠睾丸间质细胞增殖的影响。

方法

采用HE染色法观察10周龄Sox30基因野生型(Sox30

+/+

)、杂合型(Sox30

-/+

)及纯合缺失型(Sox30

-/-

)小鼠睾丸组织病理形态；采用免疫组化技术检测睾丸组织中间质细胞特异性表达蛋白3β-HSD的表达水平；采用慢病毒感染及siRNA干扰技术构建Sox30基因过表达及敲低TM3细胞模型，并在此细胞模型基础上分别采用CCK-8法、流式细胞术、RT-qPCR法以及Western blot法检测细胞存活率、细胞周期分布及周期相关基因的mRNA及蛋白表达水平。采用JASPAR数据库预测Sox30与周期相关基因的启动子区结合位点。

结果

与Sox30

+/+

和Sox30

-/+

小鼠比较，HE染色结果显示Sox30

-/-

小鼠睾丸组织中间质细胞显著增生，同时免疫组化染色发现表达3β-HSD的间质细胞增多。与Vector组相比，Sox30组TM3细胞存活率降低(

0.05)，G

期细胞周期阻滞(

0.05)，同时细胞周期相关基因CyclinD1、Cdk2等mRNA和蛋白表达水平显著降低(

0.05)。与Sox30-NC组比较，siSox30-3组TM3细胞存活率增加(

0.05)，G

期细胞周期阻滞恢复(

0.05)，同时细胞周期相关基因CyclinD1、Cdk2等mRNA和蛋白表达水平显著恢复(

0.05)。JASPAR预测发现CyclinD1、Cdk2的启动子区域均存在Sox30蛋白质可能的结合位点。

结论

Sox30基因敲除可导致小鼠间质细胞增殖异常，CyclinD1和Cdk2可能是Sox30调控细胞周期的重要靶基因。

Abstract

Objective

To investigate the effect of Sox30 gene knockout on the proliferation of mouse Leydig cells.

Methods

The pathological morphology of the testis of 10-week-old Sox30

+/+

Sox30

-/+

and Sox30

-/-

mice was observed with HE staining. Immunohistochemical assay was adopted to detect the expression level of 3β-HSD in the Leydig cells. The cell models of Sox30 gene overexpression and knockdown were constructed by lentivirus infection and siRNA interference in TM3 cell line

respectively. On the basis of these cell models

CCK-8 assay

flow cytometry

RT-qPCR and Western blotting were used to detect the cell survival rate

cell cycle distribution

and expression of cell cycle-related genes and proteins. The JASPAR database was used to predict the binding sites of Sox30 protein in promoter region of cycle related genes.

Results

Compared with Sox30

+/+

and Sox30

-/+

mice

HE staining indicated that the number of Leydig cells were significantly increased

and immunohistochemical assay showed the expression of 3β-HSD were increased the Leydig cells in the testis tissue of Sox30

-/-

mice. Compared with the blank vector group

Sox30 overexpression led to the decrease of cell count (

0.05)

cell cycle arrest in G1 phase(

0.05)

and significantly decreased expression of CyclinD1 and Cdk2 at mRNA and protein levels (

0.05). On the contrary

Sox30 knockdown resulted in opposite phenomena (

0.05). JASPAR database predicted that Sox30 may bind to the promoter of CyclinD1 and Cdk2.

Conclusion

Knockout of Sox30 induces abnormal proliferation of Leydig cells in mice. CyclinD1 and Cdk2 may be important downstream targets of Sox30 gene in the regulation of cell cycle in Leydig cells.

关键词

Keywords

references

VANDER BORGHT M, WYNS C. Fertility and infertility: Definition and epidemiology[J]. Clin Biochem, 2018, 62: 2-10.

FAINBERG J, KASHANIAN J A. Recent advances in understanding and managing male infertility[J]. F1000Res, 2019, 8: F1000 Faculty Rev-670.

XAVIER M J, SALAS-HUETOS A, OUD M S, et al . Disease gene discovery in male infertility: past, present and future[J ] . Hum Genet, 2021, 140(1): 7-19.

HART K, TADROS N N. The role of environmental factors and lifestyle on male reproductive health, the epigenome, and resulting offspring[J]. Panminerva Med, 2019, 61(2): 187-195.

曹佳, 陈卿. 生殖损害和不良妊娠结局的环境、生活方式和心理危险因素研究进展及展望[J]. 第三军医大学学报, 2021, 43(1): 1-9.

CAO J, CHEN Q. Progress and prospective of environmental, lifestyle and psychological risk factors for reproductive impairment and adverse birth outcome[J]. J Third Mil Med Univ, 2021, 43(1): 1-9.

蔡滨, 孙大林, 邓伟民, 等. 细胞周期蛋白在精子发生过程中的作用[J]. 中华男科学杂志, 2019, 25(2): 168-171.

CAI B, SUN D L, DENG W M, et al . Roles of cyclins in the progression of spermatogenesis[J ] . Natl J Androl, 2019, 25(2): 168-171.

HEINRICH A, DEFALCO T. Essential roles of interstitial cells in testicular development and function[J]. Andrology, 2020, 8(4): 903-914.

LU H M, ZHANG H S, GAO J, et al . Effects of perfluorooctanoic acid on stem Leydig cell functions in the rat[J ] . Environ Pollut Barking Essex, 2019, 250: 206-215.

CHEN X W, LI C, CHEN Y, et al . Aflatoxin B1 impairs leydig cells through inhibiting AMPK/mTOR-mediated autophagy flux pathway[J ] . Chemosphere, 2019, 233: 261-272.

YE J X, ZHANG K N, YUAN X D, et al . Di-n-Hexyl phthalate causes Leydig cell hyperplasia in rats during puberty[J ] . Toxicol Lett, 2020, 332: 213-221.

LYU Y, DONG Y Y, WANG Y Y, et al . Benzyl butyl phthalate non-linearly affects rat Leydig cell development during puberty[J ] . Toxicol Lett, 2019, 314: 53-62.

HAN F, DONG Y, LIU W B, et al . Epigenetic regulation of sox30 is associated with testis development in mice[J ] . PLoS ONE, 2014, 9(5): e97203.

HAN F, JIANG X, LI Z M, et al . Epigenetic inactivation of SOX30 is associated with male infertility and offers a therapy target for non-obstructive azoospermia[J ] . Mol Ther Nucleic Acids, 2020, 19: 72-83.

王平, 姜晓, 韩飞, 等. SOX30基因敲除对雄性小鼠睾丸组织结构及激素分泌的影响[J]. 癌变·畸变·突变, 2020, 32(5): 329-335.

WANG P, JIANG X, HAN F, et al . Effects of SOX30 gene knockout on testicular tissue structure and hormone secretion in male mice[J ] . Carcinogen Teratogen Mutagen, 2020, 32(5): 329-335.

HAN F, YIN L, JIANG X, et al . Identification of SRY-box 30 as an age-related essential gatekeeper for male germ-cell meiosis and differentiation[J ] . Aging Cell, 2021, 20(5): e13343.

ANITHA A, SENTHILKUMARAN B. Role of sox30 in regulating testicular steroidogenesis of common carp[J]. J Steroid Biochem Mol Biol, 2020, 204: 105769.

HUME S, DIANOV G L, RAMADAN K. A unified model for the G1/S cell cycle transition[J]. Nucleic Acids Res, 2020, 48(22): 12483-12501.

马帮靖, 郝翔麟, 韩飞, 等. SOX30基因调控桥粒基因DSC3抑制肺腺癌的增殖与迁移[J]. 癌变·畸变·突变, 2018, 30(3): 165-170, 175.

MA B J, HAO X L, HAN E E, et al . Inhibition of proliferation and migration of lung adenocarcinoma cells by SOX30 gene via regulation of DSC3 gene[J ] . Carcinogen Teratoge Mutagen, 2018, 30(3): 165-170, 175..

FU Q, SUN Z Y, YANG F, et al . SOX30, a target gene of miR-653-5p, represses the proliferation and invasion of prostate cancer cells through inhibition of Wnt/β-catenin signaling[J ] . Cell Mol Biol Lett, 2019, 24(1): 1-13.

SANIDAS I, MORRIS R, FELLA K A, et al . A code of mono-phosphorylation modulates the function of RB[J ] . Mol Cell, 2019, 73(5): 985-1000. e6.

WANG Q S, LI F, LIAO Z Q, et al . Low level of Cyclin-D1 correlates with worse prognosis of clear cell renal cell carcinoma patients[J ] . Cancer Med, 2019, 8(9): 4100-4109.

SINGH P, PATEL R K, PALMER N, et al . CDK2 kinase activity is a regulator of male germ cell fate[J ] . Development, 2019, 146(21): dev180273.

浏览量

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

基于基因芯片分析急性镉暴露致大鼠肝损伤及其可能机制

miR-143通过抑制AF9调控SKM-1细胞增殖和凋亡

长链非编码RNA LINC01140对急性髓系白血病U937细胞增殖及周期的影响

Hsa_circ_0000711在人肝癌细胞系中的功能研究