小鼠各期卵母细胞中SGK3 mRNA和蛋白的动态表达及其亚细胞定位

doi:10.13481/j.1671-587X.20230514

Abstract

Abstract:

Objective To discuss the expressions of serum and glucocorticoid-regulated kinase 3 （SGK3） mRNA and protein in the oocytes of the mice at different stages and their subcellular localizations，and to provide the evidence for elucidating the regulatory mechanism of SGK3 in the cell cycle of oocytes of the mice. Methods The oocytes of the mice at the germinal vesicle （GV）stage， germinal vesicle breakdown （GVBD） stage， first meiotic division （MⅠ）stage， and second meiotic division （MⅡ） stage were collected by using superovulation techniques. Real-time fluorescence quantitative PCR （RT-qPCR） and Western blotting methods were used to detect the expression levels of SGK3 mRNA and protein in the oocytes at different stages； the phosphorylation expressions of Cdc25B-Ser30 in the oocytes at different stages of the mice were detected by Western blotting method after collecting the oocytes at GV， GVBD， and MⅠ stages； the subcellular localization of SGK3 in the oocytes of the mice at different stages was observed by immunofluorescence. Results SGK3 mRNA and protein were expressed in the oocytes of the mice at all stages. Compared with GV stage， the expression levels of SGK3 mRNA and protein in the oocytes at GVBD and MⅡ stages were increased （P<0.01）. Compared with GVBD stage， the expression levels of SGK3 mRNA and protein in the oocytes at MⅠ stage were decreased （P<0.01），and the expression levels of SGK3 mRNA and protein in the oocytes at MⅡ stage were increased （P<0.01）. Compared with MⅠ stage， the expression levels of SGK3 mRNA and protein in the oocytes at MⅡ stage were increased （P<0.01）.The Cdc25B-Ser30 was dephosphorylated at GV stage and phosphorylated at GVBD and MⅠ stages.The SGK3 was localized at the nuclear membrane of the oocytes at GV stage，and entered the nucleus before GVBD stage，and localized at the nucleus during GVBD stage， distributed throughout the oocytes during MⅠ stage， and re-entered the nucleus from the cytoplasm during MⅡ stage. Conclusion SGK3 is dynamically expressed in the oocytes of the mice at different stages， and it exhibits the nuclear-cytoplasmic shuttling. The dynamic changes in the SGK3 localization may be involved in the activation of cell cycle progression.

Key words: Serum and glucocorticoid-induced protein kinase, Oocyte, Meiosis, Dynamic expression, Subcellular localization

CLC Number:

Q132

Wenning HE,Shaoqing FENG,Haiyao PANG,Jun MENG. Dynamic expressions and subcellular localization of SGK3 mRNA and protein in oocytes of mice at different stages[J].Journal of Jilin University(Medicine Edition), 2023, 49(5): 1210-1216.

Figures/Tables 3

References 21

1	孟琴琴，郭美亮，袁定芬，等. SGK1功能的研究进展及临床意义［J］. 医学综述， 2020， 26（3）： 427-432.
2	MAIYAR A C， LEONG M L L， FIRESTONE G L. Importin-alpha mediates the regulated nuclear targeting of serum- and glucocorticoid-inducible protein kinase （Sgk） by recognition of a nuclear localization signal in the kinase central domain［J］.Mol Biol Cell， 2003，14（3）： 1221-1239.
3	VASUDEVAN K M， BARBIE D A， DAVIES M A， et al. AKT-independent signaling downstream of oncogenic PIK3CA mutations in human cancer［J］. Cancer Cell， 2009， 16（1）： 21-32.
4	CHIBA K. Oocyte maturation in starfish［J］. Cells， 2020， 9（2）： 476.
5	BASNET R， GONG G Q， LI C Y， et al. Serum and glucocorticoid inducible protein kinases （SGKs）： a potential target for cancer intervention［J］. Acta Pharm Sin B， 2018， 8（5）： 767-771.
6	CHEN Y， SUN Z Y， QI M， et al. INPP4B restrains cell proliferation and metastasis via regulation of the PI3K/AKT/SGK pathway［J］. J Cell Mol Med， 2018， 22（5）： 2935-2943.
7	TESSIER M， WOODGETT J R. Role of the Phox homology domain and phosphorylation in activation of serum and glucocorticoid-regulated kinase-3［J］. J Biol Chem， 2006， 281（33）： 23978-23989.
8	ANDJELKOVIĆ M， ALESSI D R， MEIER R， et al. Role of translocation in the activation and function of protein kinase B［J］. J Biol Chem， 1997， 272（50）： 31515-31524.
9	TOSON B， FORTES I S， ROESLER R， et al. Targeting Akt/PKB in pediatric tumors： a review from preclinical to clinical trials［J］. Pharmacol Res， 2022， 183： 106403.
10	DONG Q Q， LI Z F， ZHANG H， et al. Serum and glucocorticoid-inducible kinase 3/Nedd4-2 signaling pathway participates in podocyte injury by regulating the stability of nephrin［J］. Front Physiol， 2022， 12： 810473.
11	LIAO Q Q， DONG Q Q， ZHANG H， et al. Contributions of SGK3 to transporter-related diseases［J］. Front Cell Dev Biol， 2022， 10： 1007924.
12	LANG F， COHEN P. Regulation and physiological roles of serum- and glucocorticoid-induced protein kinase isoforms［J］. Sci STKE， 2001， 2001（108）： re17.
13	NTOSTIS P， ILES D， KOKKALI G， et al. The impact of maternal age on gene expression during the GV to MⅡ transition in euploid human oocytes［J］. Hum Reprod， 2021， 37（1）： 80-92.
14	YOON S Y. Role of type 1 inositol 1， 4， 5-triphosphate receptors in mammalian oocytes［J］. Dev Reprod， 2019， 23（1）： 1-9.
15	QI L B， LIU B X， CHEN X， et al. Single-cell transcriptomic analysis reveals mitochondrial dynamics in oocytes of patients with polycystic ovary syndrome［J］. Front Genet， 2020， 11： 396.
16	AHMED M， FEZAI M， UZCATEGUI N L， et al. SGK3 sensitivity of voltage gated K⁺ channel Kv1.5 （KCNA5）［J］. Cell Physiol Biochem， 2016， 38（1）： 359-367.
17	LANG F， BÖHMER C， PALMADA M， et al. （Patho）physiological significance of the serum- and glucocorticoid-inducible kinase isoforms［J］. Physiol Rev， 2006， 86（4）： 1151-1178.
18	HIRAOKA D， HOSODA E， CHIBA K， et al. SGK phosphorylates Cdc25 and Myt1 to trigger cyclin B-Cdk1 activation at the meiotic G2/M transition［J］. J Cell Biol， 2019， 218（11）： 3597-3611.
19	JU C， ZHOU R H， SUN J， et al. LncRNA SNHG5 promotes the progression of osteosarcoma by sponging the miR-212-3p/SGK3 axis［J］. Cancer Cell Int， 2018， 18： 141.
20	FAN X R， ZHANG Z Y， WANG R H， et al. miR-376a functions as tumor suppressor by targeting SGK3 in renal cell carcinoma［J］. Eur Rev Med Pharmacol Sci， 2019， 23（9）： 3726-3732.
21	ZHENG H Z， WU L P， WANG X D， et al. Risk of nasopharyngeal carcinoma associated with single nucleotide polymorphisms in the microRNA binding site of SGK3［J］. Genet Test Mol Biomarkers，2020，24（8）： 508-519.

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Dynamic expressions and subcellular localization of SGK3 mRNA and protein in oocytes of mice at different stages

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