&nbsp;Lentiviral vector mediated steroidogenic factor-1 silencing regulating decidualization and autophagy of endometrial stromal cells

CHEN Xiao-Yan; REN Jian

doi:10.16098/j.issn.0529-1356.2024.02.009

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Acta Anatomica Sinica ›› 2024, Vol. 55 ›› Issue (2) : 188-195. DOI: 10.16098/j.issn.0529-1356.2024.02.009

Lentiviral vector mediated steroidogenic factor-1 silencing regulating decidualization and autophagy of endometrial stromal cells

REN Jian^1,2 CHEN Xiao-yan^3*

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Abstract

Objective To explore the influence and mechanism of lentiviral vector-mediated steroidogenic factor-1 (SF-1) silence on decidual process of human endometrial stromal cells (hESCs). Methods The endometrial tissues of patients with endometriosis (EM) and normal pregnant women were collected, and the differential expression of SF-1 was detected by Real-time PCR and immunohistochemical staining. hESCs were isolated from the endometrial tissue of normal pregnancy and identified, hESCs were divided into control group, estradiol (E₂) + progesterone (P4) group, short hairpin RAN(shRNA, sh)-normal control(NC)+E₂+P4 group, sh-SF-1+E₂+P4 group, after the corresponding processing, the morphological changes of hESCs were observed under an inverted microscope. Real-time PCR and Western blotting were used to detect the mRNA expression levels and protein levels of human insulin-like growth factor-binding protein-1 (IGFBP-1) and prolactin (PRL) in cells, respectively, flow cytometry was used to determine the cycle distribution of the cells, immunofluorescence staining was used to observe the expression of the intracellular auto-bid microtubule-associated protein light chain 3(LC3), Western blotting was used to determine the protein level of intracellular autophagy-related proteins LC3-Ⅱ, LC3-Ⅰ and Beclin-1. Results The relative expression of SF-1 mRNA and the positive rate of SF-1 protein in endometrial tissue of EM patients were higher than those of normal pregnancy endometrial tissue (P<0.05). Compared with the sh-NC+E₂+P4 group, the cells in the sh-SF-1+E₂+P4 group were mostly long-spindle, and there was no obvious decidualization. The relative mRNA expression and protein expression of IGFBP1 and PRL were significantly down-regulated (P<0.05), the proportion of cells in G₀/G₁ phase was significantly decreased, the proportion of cells in S phase was significantly increased (P<0.05), the ratio of LC3-Ⅱ/LC3-Ⅰ was significantly increased, and the relative expression of Beclin-1 protein was also significantly up-regulated (P<0.05). Conclusion The expression of SF-1 in the endometrial tissue of EM patients is increased, and SF-1 silencing mediated by lentiviral vector can inhibit the decidualization process of hESCs, which may be related to regulating the level of autophagy.

Key words

Endometrial stromal cell / Steroidogenic factor-1 / Decidualization / Autophagy / Western blotting / Human

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REN Jian CHEN Xiao-yan. Lentiviral vector mediated steroidogenic factor-1 silencing regulating decidualization and autophagy of endometrial stromal cells[J]. Acta Anatomica Sinica. 2024, 55(2): 188-195 https://doi.org/10.16098/j.issn.0529-1356.2024.02.009

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