Abstract
Purpose
Endometriosis, a gynecological disease, is difficult to be cured. Currently, to identify more potential biomarkers for the early diagnosis of endometriosis is urgently needed. Insulin like growth factor 2 (IGF2) has been revealed to correlate with endometriosis. This research aimed to further explore the role of IGF2 and its up-stream mechanism in endometriosis.
Methods
Primary ectopic endometrial stromal cells (EESCs) were extracted from ectopic endometrial tissues which were pathological endometrial tissues resected from three patients with II-III endometriosis. Primary normal endometrial stromal cells (NESCs) were extracted from normal endometrial tissues of two patients with grade III cervical dysplasia and one patient with uterine leiomyoma III. Four endometriotic cell lines (EEC145T, hEM15A, hEM5B2, and 12Z) and normal human endometrial epithelial cells (hEECs) were purchased. Cell proliferation, migration, and invasion were evaluated through functional assays. The molecular interaction between RNAs was investigated through mechanistic analyses.
Results
We discovered that IGF2 was upregulated in purchased endometriotic cells and primary EESC. Suppression of IGF2 hampered cell proliferation, migration, and invasion. Furthermore, insulin-like growth factor 2 antisense RNA (IGF2-AS) was uncovered to positively regulate IGF2 expression and enhanced proliferative, migratory, and invasive abilities of endometriotic cells. Mechanistically, miR-370-3p was found to bind with IGF2-AS and IGF2. IGF2-AS competitively bind with miR-370-3p to upregulate IGF2. Furthermore, IGF2-AS was revealed to activate the PI3K/AKT/mTOR signaling pathway through targeting miR-370-3p/IGF2 axis.
Conclusion
IGF2-AS promotes endometriotic cell growth via regulating IGF2/miR-370-3p axis and further activating PI3K/AKT/mTOR signaling pathway.
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Data availability
Research data are not shared.
Abbreviations
- lncRNA:
-
Long non-coding RNA
- IGF2-AS:
-
Insulin like growth factor 2 antisense RNA
- ceRNA:
-
Competing endogenous RNA
- miRNA:
-
MicroRNA
- IGF2:
-
Insulin like growth factor 2
- qRT-PCR:
-
Quantitative real-time PCR
- CCK-8:
-
Cell counting kit-8
- EdU:
-
5-Ethynyl-2′-deoxyuridine
- RIP:
-
RNA immunoprecipitation
- shRNA:
-
Short hairpin RNA
- FBS:
-
Fetal bovine serum
- GAPDH:
-
Glyceraldehyde-3-phosphate dehydrogenase
- PVDF:
-
Polyvinylidene fluoride
- DMEM:
-
Dulbecco’s modified Eagle’s medium
- SD:
-
Standard deviation
- NC:
-
Negative control
- ANOVA:
-
Analysis of variance
- ATCC:
-
American Type Culture Collection
- OD:
-
Optical density
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Jin, X., Feng, J. & Cheng, X. LncRNA IGF2-AS promotes endometriosis progression through targeting miR-370-3p/IGF2 axis and activating PI3K/AKT/mTOR signaling pathway. J Assist Reprod Genet 39, 2699–2710 (2022). https://doi.org/10.1007/s10815-022-02638-2
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DOI: https://doi.org/10.1007/s10815-022-02638-2