Effect of the transcription factor SOX4 on the proliferation， migration and apoptosis of ovarian granulosa cells
10.19405/j.cnki.issn1000-1492.2023.10.006
- Author:
Mengmeng Fan
1
;
Cong Li
1
;
Xue Liang
1
;
Wanqiu Zhang
1
;
Huiping Wang
1
;
Zhimin Zhai
1
Author Information
1. Dept of Hematology，The Second Affiliated Hospital of Anhui Medical University，Hefei 230601
- Publication Type:Journal Article
- Keywords:
SOX4;
ovary granulosa cells;
proliferation;
migration;
apoptosis
- From:
Acta Universitatis Medicinalis Anhui
2023;58(10):1646-1651
- CountryChina
- Language:Chinese
-
Abstract:
Objective : To investigate the impact of SOX4 on ovarian granulosa cells，stable overexpression of SOX4 was achieved in human KGN cell line，followed by analysis of its effects on proliferation，migration and apoptosis.
Methods : The recombinant lentiviral plasmid pLV-EF1a-GFP / Puro-SOX4 was generated through homologous recombination with linearized pLV-EF1a-GFP / Puro vector．Human ovarian granulosa cells ( KGN cell line ) were transduced with Lentiviral expression vectors．KGN cells infected with pLV-EF1a-GFP / Puro-NC were served as the LV-CON group，while those infected with pLV-EF1a-GFP / Puro-SOX4 were designated as the LV-SOX4 group．Following transfection，puromycin selection was employed to establish stable SOX4-expressing KGN cells．The expres- sion levels of SOX4 m RNA and protein in KGN cells from the LV-CON and LV-SOX4 groups were assessed using RT-qPCR and Western blot analysis．Cell proliferation was assessed using the CCK-8 assay in both LV-CON and LV-SOX4 groups．Cell migration ability was evaluated by means of a cell scratch test in these two groups．The proportion of apoptotic cells was determined via flow cytometry analysis in both LV-CON and LV-SOX4 groups.
Results:The sequencing results of pLV-EF1a-GFP / Puro-SOX4 indicated a complete match between the inserted gene se- quence and the SOX4 mRNA sequence．The lentiviral titers were 7 × 108 TU / ml in the LV-CON group and 1 × 108 TU / ml in the LV-SOX4 group．The recombinant plasmid was successfully transfected into KGN cells with a transfection efficiency of over 90% under fluorescence inverted microscopy．The results of RT-qPCR and Western blot tests demonstrated a significant increase in the expression level of SOX4 in KGN cells of LV-SOX4 group compared to that of LV-CON group (t = 3. 10，P ＜0. 05 ; t = 14. 20，P ＜0. 05) ．The CCK-8 assay results demonstrated that the LV-SOX4 group exhibited a significant increase in cell proliferation (24 h : t = 45. 92，P＜0. 01 ; 72 h : t = 25. 60，P ＜0. 01) compared to the LV-CON group．The cell scratch assay indicated that the migratory capacity of KGN cells in the LV-SOX4 group was significantly enhanced (t = 7. 65，P ＜0. 01) compared to that in the LV-CON group. The LV-SOX4 group exhibited a significant reduction in apoptosis ratio (t = 25. 84，P＜0. 01) compared to the LV- CON group．
Conclusion :SOX4-overexpressing KGN cell line was successfully established，and the overexpression of SOX4 facilitated proliferation and migration while inhibiting apoptosis in human ovarian granulosa cells.
- Full text:2024071217461067170转录因子SOX4对卵巢颗粒细胞增殖、迁移及凋亡的影响_范蒙蒙.pdf
