Hsa_circ_0008726 regulates the proliferation, migration, and invasion of trophoblast cells in preeclampsia through modulating the miR-1290-LHX6 signaling pathway

Abstract

Background: Preeclampsia (PE) is a serious complication of pregnancy, with a global incidence of about 2%-8%. It is one of the important causes of morbidity and mortality among the pregnant women and perinatal infants. Circular RNA (circRNA) has been confirmed to play an important regulatory role in PE. This study aimed to evaluate the role of hsa_circ_0008726 in the occurrence and development of PE.

Methods: The expression of hsa_circ_0008726 in PE placental tissue and blood was detected by qRT-PCR. CCK-8, wound closure, and Transwell assay were used to measure cell proliferation, migration, and invasion. Bioinformatics prediction, Western blotting, and dual-luciferase reporter gene detection were used to explore the mechanism of hsa_circ_0008726 in HTR8/SVneo cells.

Results: The expression level of circ_0008726 in the placental tissue and blood samples of PE patients was significantly higher than that of normal controls. The overexpression of circ_0008726 can significantly inhibit the proliferation, migration, and invasion ability of HTR-8/SVneo cells. While the silence of circ_0008726 showed an opposite effect. Furthermore, hsa_circ_0008726 can modulate the expression of LHX6 by adsorbing miR-1290.

Conclusion: Hsa_circ_000872 can regulate LHX6 by adsorbing miR-1290 to inhibit PE progression, thus establishing hsa_circ_000872 as a potential target for predicting and treating PE.

Keywords: LHX6; hsa_circ_0008726; miR-1290; preeclampsia.

FIGURE 1

circ_0008726 was highly expressed in the tissues and blood of PE. (A) qRT‐PCR was employed to examine the expression of circ_0008726 in the placenta of PE patients and normal pregnant women, N = 3. (B) qRT‐PCR was used to detect the expression of circ_0008726 in the blood samples of PE patients and normal pregnant women, N = 17. (C) circ_0008726 is formed by back‐splicing of exons 4, 5, and 6 of gene DNAJB6. (D) qRT‐PCR was used to determine the knockdown efficiency of circ_0008726 in HTR‐8/SVneo cells. (E) qRT‐PCR was used to analyze the overexpression efficiency of circ_0008726 in HTR‐8/SVneo cells. *p < 0.05, **p < 0.01, ***p < 0.001

FIGURE 2

circ_0008726 inhibited the proliferation, migration, and invasion of trophoblast cells (A) The viability of HTR‐8/SVneo cells was analyzed by CCK‐8 assay. (B) The migration of HTR‐8/SVneo cells was determined by wound healing test. (C) The invasion ability of HTR‐8/SVneo cells was analyzed by Transwell invasion. **p < 0.01, ***p < 0.001

FIGURE 3

miR‐1290 can serve as a potential target of circ_0008726. (A) The database predicted that there was a potential binding miRNA with circ_0008726. (B) The predictive analysis of the binding site of miR‐1290 and circ_0008726; (C) luciferase reporter gene detection for HEK239T luciferase activity in cells; (D) luciferase reporter gene assay was carried out to detect luciferase activity in HTR‐8/SVneo cells; (E) qRT‐PCR was used to detect the expression level of circ_0008726 in HTR‐8/SVneo cells. *p < 0.05, **p < 0.01

FIGURE 4

LHX6 can be targeted by miR‐1290. (A and B) qRT‐PCR was employed to detect the expression level of LHX6 in HTR‐8/SVneo cells; (C and D) Western Blot was used to analyze the expression level of LHX6 protein in HTR‐8/SVneo cells. *p < 0.05, **p < 0.01

FIGURE 5

circ_0008726 suppressesd HTR8/SVneo cell growth, migration, and invasion by targeting miR‐1290. (A) The viability of HTR‐8/SVneo cells was analyzed by CCK‐8 assay. (B) The migratory potential of HTR‐8/SVneo cells was determined by wound healing test. (C) The invasion ability of HTR‐8/SVneo cells was examined by Transwell assay. *p < 0.05, **p < 0.01, ***p < 0.001