Role of Notch 1 signaling and glycolysis in the pathogenic mechanism of adenomyosis

doi:10.12122/j.issn.1673-4254.2024.08.19

Abstract

Abstract:

Objective To investigate the expressions of glycolysis-related factors and changes in Notch1 signaling in endometrial tissues of adenomyosis (AM) and Ishikawa cells to explore the pathogenesis of AM. Methods Eutopic endometrial tissues were collected from 8 patients with AM and 8 patients with uterine fibroids matched for clinical characteristics (control group). The expressions of Notch1 signaling proteins and glycolysis-related factors in the collected tissues were detected using qRT-PCR and Western blotting, and the levels of glucose and lactic acid were determined. An Ishikawa cell model with lentivirus-mediated stable Notch1 overexpression was established for assessing cell survival rate with CCK-8 assay, cell migration and invasion abilities with Transwell migration and invasion assays, and glycolytic capacity by determining the extracellular acidification rate. Results Compared with those in the control group, the endometrial tissues in AM group showed significantly increased expression level of carbohydrate antigen 125 (CA125), increased mRNA expression levels of Notch1, HK2 and PDHA and protein expressions of Notch1, GLUT1, HK2, PKM and PDHA, lowered glucose level and increased lactate level. The Ishikawa cell models with stable Notch1 overexpression exhibited significantly increased cell survival rate with attenuated cell migration and invasion abilities and decreased glycolysis capacity and reserve. Conclusion The Notch1 signaling pathway participates in the pathogenesis of AM possibly by regulating the proliferation, migration, invasion and glycolysis of endometrial cells.

Key words: adenomyosis, endometrium, glycolysis, Ishikawa, lentivirus

Xiaohui WEN, Shiya HUANG, Xuehong LIU, Kunyin LI, Yongge GUAN. Role of Notch 1 signaling and glycolysis in the pathogenic mechanism of adenomyosis[J]. Journal of Southern Medical University, 2024, 44(8): 1599-1604.

Figures/Tables 9

References 32

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Primer	Sequence (5' to 3')
Notch1	TGGACCAGATTGGGGAGTTC
Notch1	GCACACTCGTCTGTGTTGAC
GLUT1	TGGCATCAACGCTGTCTTCT
GLUT1	AGCCAATGGTGGCATACACA
HK2	GTGAATCGGAGAGGTCCCAC
HK2	GCTAACTTCGGCCACAGGAT
PKM	ATGTCGAAGCCCCATAGTGAA
PKM	TGGGTGGTGAATCAATGTCCA
PDHA	ATGGAATGGGAACGTCTGTTG
PDHA	CCTCTCGGACGCACAGGATA
GAPDH	CAGGAGGCATTGCTGATGAT
GAPDH	GAAGGCTGGGGCTCATTT

Primer	Sequence (5' to 3')
Notch1	TGGACCAGATTGGGGAGTTC
Notch1	GCACACTCGTCTGTGTTGAC
GLUT1	TGGCATCAACGCTGTCTTCT
GLUT1	AGCCAATGGTGGCATACACA
HK2	GTGAATCGGAGAGGTCCCAC
HK2	GCTAACTTCGGCCACAGGAT
PKM	ATGTCGAAGCCCCATAGTGAA
PKM	TGGGTGGTGAATCAATGTCCA
PDHA	ATGGAATGGGAACGTCTGTTG
PDHA	CCTCTCGGACGCACAGGATA
GAPDH	CAGGAGGCATTGCTGATGAT
GAPDH	GAAGGCTGGGGCTCATTT

Group	Age (year)	Uterine volume (cm³)	Number of pregnancies	Number of abortions	CA125 (U/mL)
CON	46.63±2.78	280.38 (128.75-1009.46)	3.00 (2.00-3.00)	0.00 (0.00-1.75)	14.44 (12.16-23.82)
AM	44.88±2.95	636.08 (396.14-1020.82)	3.50 (2.25-5.00)	1.50 (0.25-3.50)	45.01 (28.51-73.14)**

Group	Age (year)	Uterine volume (cm³)	Number of pregnancies	Number of abortions	CA125 (U/mL)
CON	46.63±2.78	280.38 (128.75-1009.46)	3.00 (2.00-3.00)	0.00 (0.00-1.75)	14.44 (12.16-23.82)
AM	44.88±2.95	636.08 (396.14-1020.82)	3.50 (2.25-5.00)	1.50 (0.25-3.50)	45.01 (28.51-73.14)**

Group	Notch1	GLUT1	HK2	PKM	PDHA
CON	1.31±0.78	0.71 (0.65-1.13)	1.01 (0.50-5.26)	1.01 (0.29-3.63)	1.78±1.68
AM	5.12±3.46*	3.04 (1.21-3.85)	3.31 (1.58-12.38)*	3.10 (1.40-8.02)	6.92±5.05*