| Background:Endometriosis（EMs）is an estrogen-dependent disease that occurs in women of childbearing age,and its pathogenesis is not yet clear.Although the disease is a benign disease,it has the characteristics of malignant tumor invasion and metastasis.nm23-H1 is an important tumor metastasis suppressor gene and is regulated by estrogen.nm23-H1 can reduce the expression of MMP-2/9 and enhance the expression of TIMP by the RAS-RAF-MEK-ELK1 pathway,the P3K-AKT-NF-κB/AP-1 pathway or the RAS-RAC-MKK3/6-p38 pathway in inhibiting tumor invasion and metastasis.eventually weakening the metastasis of tumor cells.Studies have shown that that the expression of MMP-2/9（MMP-2/9）in ectopic endometrium of EMs has a significant change.To further clarify the correlation between the occurrence and development of nm23-H1 and EMs and whether it can regulate the expression of MMP-2/9 expression promotes the development of EMs.In this study,the correlation between MMP-2,MMP-9 gene expression and nm23-H1 and EMs and their specific mechanism were explored by RT-PCR,cell culture and siRNA transfection.Through the above experimental study,the role of nm23-H1 in the occurrence of EMs will be basically clarified,trying to clarify the specific role of the pathway,providing a new target for the treatment of EMs.Objective:To explore the relationship between nm23-H1 and endometriosis,trying to explore the role of nm23-H1-mediated MMP-2/MMP-9 expression in the pathogenesis of EMs.Method:Take normal endometrial tissue and endometriosis ectopic tissue in 10 cases,after digestion,separation and culture of endometrial stromal cells.The cells were stimulated with 10-6mol/L 17β-estradiol（C18H24O2,272.38）.Then harvested cells and extracted the RNA.Detected the expressions of nm23-H1,MMP-2 and MMP-9 by RT-PCR,to explore the effect of estrogen on the expression of nm23-H1,MMP-2 and MMP-9.With siRNA transfected cells to silenced the expression of nm23-H1.Then extracted RNA and detected the expression of nm23-H1,MMP-2 and MMP-9,to explore whether the low expression of nm23-H1 affect the expression of MMP-2,MMP-9.Result:Regular qRT-PCR to detect gene expression:1.Genes expression without any treatment（1）The mRNA expression levels of nm23-H1 in the stromal cells of the the endometriosis group and the normal control group were 0.61±0.10 and 1.03±0.24,respectively.Compared with the two groups,the mRNA expression of nm23-H1 in endometriosis group was lower than that in the normal control group（t=4.58,P=0.00,P<0.05）.（2）The mRNA expression levels of MMP-2 in the stromal cells of the endometriosis group and the normal control group were 1.17±1.39,6.87±8.96,respectively.There was no statistically significant difference between the two groups（t=1.77,P=0.10,P>0.05）.（3）The mRNA expression levels of MMP-9 in the stromal cells of the endometriosis group and the normal control group were 22.38.±21.37,2.94±5.04,respectively.Compared with the two groups,the mRNA expression of MMP-9gene in endometriosis group was higher than that in normal control group（t=-2.52,P=0.04,P<0.05）.2.Gene expression after estrogen stimulation（1）Normal group:（1）The expression level of nm23-H1 mRNA in normal cells before and after estrogen treatment was 1.02±0.24,0.57±0.23,respectively.Compared with the two groups,the mRNA expression of nm23-H1 after estrogen treatment was lower than that before estrogen treatment,and the difference was statistically significant（t=5.33,P=0.000,P<0.05）.（2）The expression levels of MMP-2 mRNA in normal cells before and after estrogen treatment were 6.87±8.96 and 7.71±14.84,respectively.There was no significant difference between the two groups（t=-0.15,P=0.88,P>0.05）.（3）mRNA expression levels of MMP-9 in stromal cells before and after estrogen treatment in the normal group were 2.94±5.04,2.14±4.98.There was no significant difference between the two（t=0.36,P=0.72,P>0.05）.（2）Endometriosis:（1）The mRNA expression levels of nm23-H1 gene before and after estrogen treatment in endometriosis were 1.01±0.17 and 0.49±0.26,respectively.Compared with the two groups,the expression level of nm23-H1 mRNA after estrogen treatment was lower than before estrogen treatment,and the difference was statistically significant（t=4.72,P=0.000,P<0.05）.（2）The mRNA expression levels of MMP-2 gene in the stromal cells before and after estrogen treatment were 2.021±2.39 and 5.21±4.79,respectively.There was no significant difference between the two groups（t=-1.69,P=0.11,P>0.05）.（3）The mRNA expression levels of MMP-9 in the stromal cells before and after estrogen treatment were significantly different.It is 2.24±2.01,2.53±2.12.There was no significant difference between the two（t=-0.28,P=78,P>0.05）.3.Genes expression after siRNA transfection（1）Normal group:（1）mRNA expression levels of nm23-H1 gene in the stromal cells before and after transfection of the normal group were 1.03±0.27,0.11±0.08,respectively.The mRNA expression of nm23-H1 gene after siRNA transfection was lower than that before siRNA transfection,the difference was statistically significant（t=10.32,P=0.000,P<0.05）.（2）mRNA expression levels of MMP-2 mRNA in stromal cells before and after transfection of normal group were 8.00±13.08 and 10.45±17.49,respectively.The difference between the two was not statistically significant（t=-0.36,P=0.72,P>0.05）.（3）mRNA expression levels of MMP-9 gene in normal cells before and after transfection were 2.46±4.53 and 5.26±6.28,respectively.There was no significant difference between the two groups（t=-1.14,P=0.27,P>0.05）.（2）Endometriosis group:（1）The mRNA expression level of nm23-H1 gene before and after siRNA transfection in endometriosis group was 1.31±1.23,0.01±0.00,respectively.The mRNA expression of nm23-H1 gene after siRNA transfection was lower than that before siRNA transfection（t=2.57,P=0.02,P<0.05）.（2）The expression levels of MMP-2 mRNA in the mesenchymal cells before and after siRNA transfection were 24.5±7.57 and 2.27±4.65,respectively.There was no significant difference between the two groups（t=0.74,P=0.48,P>0.05）.（3）The mRNA expression levels of MMP-9 gene in the mesenchymal cells before and after siRNA transfection were all 26.84±62.62 and 3.63±6.55,respectively.There was no significant difference between the two groups（t=-1.04,P=0.33,P>0.05）.Conclusion:1.Compared with the normal control group,the nm23-H1 gene was lowly expressed in the endometriosis group and may participate in the occurrence and development of endometriosis.2.Estrogen stimulation can down-regulate the expression of nm23-H1 gene.3.By silencing the nm23-H1 gene expression,MMP2/9 gene expression did not change significantly. |
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