Objective To explore the effect of TAM family receptor tyrosine kinases (Axl) on signaling pathway of mammalian target of rapamycin (mTOR) in patients with endometriosis (EMS).
Methods The eutopic and ectopic endometrium tissues of 5 patients with ovarian EMS were collected, and 5 normal endometrium tissues were selected as controls. Human ectopic endometrial stromal cells (HEcESCs), human eutopic endometrial stromal cells (HEuESCs) and human normal endometrial stromal cells (HEnESCs) were isolated and cultured for immunohistochemical identification, and the mechanism of regulating PI3K/AKT signal pathway by targeting upstream Axl and mTOR kinase at cell level to affect apoptosis of human embryonic stem cells (HESCs) was analyzed. Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to verify the differences of Axl, mTOR mRNA expressions in HEcESCs, HEuESCs and HEnESCs; at the cell level, siRNA transfection was used to regulate Axl targets, rapamycin was used to regulate mTOR targets, and Western blotting and qRT-PCR were used to detect the differences in the expressions of Axl, mTOR, phosphorylated mTOR (p-mTOR) and CyclinD1.
Results Compared with HEnESCs, the expression levels of Axl mRNA and mTOR mRNA in HEuESCs and HEcESCs showed significant difference (P < 0.05); compared with the blank control group, the expression levels of Axl mRNA and CyclinD1 mRNA in rapamycin treated group and Axl-siRNA treated group showed significant difference (P < 0.05); compared with the rapamycin treated group, the Axl mRNA expression level in the Axl-siRNA treated group showed significant difference (P < 0.05); compared with the negative control group, the expression levels of Axl mRNA and CyclinD1 mRNA in the rapamycin treated group showed significant difference (P < 0.05); compared with the negative control group, the expressions of Axl mRNA and CyclinD1 mRNA in the Axl-siRNA treated group showed significant difference (P < 0.05); compared with the blank control group, the relative expression level of mTOR mRNA in the rapamycin treated group showed significant difference (P < 0.05). There was no significant difference in mTOR protein expression among the four groups (P>0.05); compared with the blank control group, the expressions of p-mTOR and CyclinD1 protein in the rapamycin treated group and Axl-siRNA treated group showed significant difference (P < 0.05), and the difference compared with the negative control group was also statistically significant (P < 0.05).
Conclusion Regulation of Axl and mTOR can promote apoptosis of HEcESCs, and Axl participates in the occurrence of EMS by mediating mTOR signal pathway.
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