The Role And Significance Of LASP1 Protein In Adenomyosis

Objective:In the previous study group's success screened adenomyosis reign and differentially expressed protein in ectopic endometrial stromal cells,the present study in which the differentially expressed protein LASP1(LIM and SH3 protein 1)by,TGM2(transglutaminase 2)and PRDX4(peroxiredoxin 4)performed RT-PCR(reverse transcription polymerase chain reaction)and WB(western blot)verification;LASP1 gene promoter demethylation condition check and low expression of the gene after LASP1 ectopic endometrial stromal cells biological function change,and to explore LASP1 proteins differentially expressed between adenomyosis ectopic endometrial stromal cells the role and significance,in order to further explore the pathogenesis of adenomyosis provide theoretical support.Methods:Select the line due to adenomyosis hysterectomy / total resection surgery myomectomy or uterine gland,whichever eutopic and ectopic endometrium samples(all specimens confirmed through histopathology)as the experimental group,all patients with no other merger parts of the tumor,preoperative three months were not receiving hormone therapy and anti-inflammatory or IUD.And all patients in the proliferative phase endometrium.Tubal obstruction and infertility patients hysteroscopy line,whichever endometrial sample as a control group.After digestion,filters,differential adherence and other isolated and purified culture ectopic reign inter endometrial stromal cells and control endometrial stromal cells;RT-PCR and WB were detected LASP1-mRNA and protein expression;the methylation status of flight mass spectrometry to detect LASP1 gene promoter region;Construction LASP1-shRNA lentivirus interference,and transfected into ectopic stromal cells established pSilencer-LASP1 cells;CCK-8(cell counting kit)was used to detect cell proliferation,Transwell assay cell migration and invasion,flow cytometry PE /7-AAD double staining cells apoptosis situation.Results:(1)Immunohistochemistry showed: LASP1 protein in adenomyosis ectopic tissue organization and expression levels were significantly higher than the reign of endometrial tissue in control,the difference was statistically significant(P <0.05);(2)Isolated and purified cultured cells chemical identification vimentin(wave protein)by immune cells were positive,cytokeratin(keratin)negative;(3)RT-PCR and WB test results showed: LASP1-mRNA and protein in ectopic within LASP1 membrane between stromal cells expression were significantly higher than the control endometrial stromal cells,the difference was statistically significant(P <0.05);(4)Time of flight mass spectrometry showed that: with the control phase of endometrial stromal cells ratio,adenomyosis ectopic endometrial stromal cells LASP1 gene promoter region LASP1-5F-CpG site 14.15.16 island demethylation situation exists,the difference was statistically significant(P <0.05);(5)Compared with the negative control group of ectopic endometrial stromal cells,proliferation of low gene expression LASP1 ectopic endometrial stromal cells,migration and invasion were significantly weakened,the differences were statistically significant(P <0.05);but inhibition of apoptosis gene expression differences after LASP1 ectopic endometrial stromal cells was not statistically significant(P> 0.05).Conclusion:Adenomyosis uterus ectopic endometrial stromal cells LASP1 gene promoter CpG island methylation status to the presence of a partial,its mRNA and protein expression were significantly up-regulated water;reduce LASP1 ectopic endometrial stromal cells expression proliferation,migration and invasion were significantly decreased,but no significant effect on apoptosis.LASP1 showed high expression in adenomyosis uterus ectopic endometrial stromal cells may be related to the promoter CpG island methylation of the relevant part to,and involved in the development process of adenomyosis,but also its mechanisms for further study.