| Objective:To explore the expression of nm23-H1 gene and MT1-MMP gene in endometriosis,to understand the correlation between the occurrence of endometriosis and nm23-H1 gene and MT1-MMP gene,and to provide new ideas for clinical diagnosis,prevention and treatment of endometriosis.Methods:After digestion and isolation of 10 normal endometrium and 10 ectopic tissues,the primary cultured stromal cells were stimulated by adding 0,10-8 mol/L and10-5 mol/L of 17 beta-estradiol?17beta-Estradiolum?to the cells of the ectopic group after adherence.After 48 hours,the cells were collected and the total RNA was extracted by Trizol reagent,and the reverse transcription was performed to generate the DNA.The normal cells were detected by qRT-PCR.The expression of nm23-H1gene and MT1-MMP gene in ectopic endometrial cells was also studied.Normal and ectopic endometrium were taken to obtain endometrial glandular epithelial cells by frozen section,HE kit staining and laser capture microdissection.After RNA was extracted by microkit,it was retranscribed into cDNA.The expression of nm23-H1gene and MT1-MMP gene in normal and ectopic endometrial epithelial cells was detected by Q200 droplet digital PCR.Result:1.The expression levels of nm23-H1 gene in primary cultured stromal cells of endometriosis tissue and normal endometrium tissue were 0.321±0.111 and0.978±0.256,respectively.The expression levels of nm23-H1 gene in endometriosis were lower than those in normal endometrium control group,P=0.026,with significant difference.2.The expression levels of MT1-MMP gene in primary cultured stromal cells of endometriosis and normal endometrium were 1.438±0.277 and 0.508±0.141,respectively.Compared with normal endometriosis,the expression level of MT1-MMP gene in endometriosis was higher than those in normal endometrium control group?P=0.011?.The difference was statistically significant.3.The expression of nm23-H1 gene in primary cultured stromal cells after digestion of endometriosis tissue was 1.028±0.189,0.553±0.204 and 0.271±0.099 respectively after stimulation with different concentrations of estrogen at 0,10-8 mol/L and 10-5mol/L.With the increase of estrogen concentration,the expression level of nm23-H1gene in endometriosis stromal cells became lower and lower,there were differences between estrogen-free group and estrogen-stimulated group with 10-5 mol/L concentration.The expression of MT1-MMP gene was 1.599±0.722,4.580±2.240 and3.383±1.784,respectively.The expression of MT1-MMP gene was increased in the estrogen-stimulated group,with the highest expression in the 10-5 mol/L concentration group,but there was no significant difference among the groups.4.The expression of nm23-H1 gene in normal endometrium and endometriotic epithelial cells was 0.097±0.019 and 0.048±0.008,respectively.The expression level of nm23-H1 gene in endometriosis was lower than that in normal endometrium control group,P=0.049,with statistically difference.5.The expression of MT1-MMP gene in normal endometrium and endometriotic epithelial cells was 0.219±0.041 and 0.416±0.062,respectively.Compared with the normal endometriosis group,the expression of MT1-MMP gene in endometriosis group was higher than that in normal endometrium control group,P=0.017,the difference was statistically significant.Conclusion:1.Compared with the primary cultured stromal cells of normal endometrial tissue,the expression of nm23-H1 gene in primary cultured stromal cells of endometriosis tissue decreased,while the expression of MT1-MMP gene increased.This is consistent with the previous research on endometriosis tissue.Nm23-H1 is involved in the occurrence of endometriosis as a tumor suppressor gene.Whether the expression of nm23-H1 gene will affect MT1-MMP and lead to endometriosis remains to be further explored.2.With the increase of estrogen concentration,the expression of nm23-H1 gene has a downward trend,while the expression of MT1-MMP gene has no obvious upward or downward trend.Follow-up can increase the sample size,optimize the cell culture program,and further study to clarify the relationship between the two.3.The expression of nm23-H1 gene in endometrial epithelial cells captured by laser was decreased and MT1-MMP gene was increased compared with normal control group by dd-PCR.Epithelial cells and stromal cells may play a common role in endometriosis. |
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