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MiR-183 Regulates ITGB1P Expression and Promotes Invasion of Endometrial Stromal Cells.

Chen J, Gu L, Ni J, Hu P, Hu K, Shi YL - Biomed Res Int (2015)

Bottom Line: We applied in the previous study miRNA microarray screening analysis to identify several differentially expressed miRNAs, including miR-183 in normal, eutopic, and ectopic endometrium.Western blotting analyses showed that integrin β1 (ITGB1), but not AMIGO2, was affected by miR-183 overexpression, whereas no protein expression of VAV3 and PSEN2 was detected.Moreover, we found that ITGB1 is overexpressed in the endometrium of endometriosis patients.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Reproductive Medicine, Department of Gynecology, Nanjing Maternity and Child Health Care Hospital Affiliated to Nanjing Medical University, 123 Tianfei Road, Nanjing, Jiangsu 210029, China.

ABSTRACT
We applied in the previous study miRNA microarray screening analysis to identify several differentially expressed miRNAs, including miR-183 in normal, eutopic, and ectopic endometrium. Knockdown of miR-183 expression induced the invasiveness and inhibition of apoptosis in endometrial stromal cells. The current study aims to identify the miR-183 targets with relevance to cell functions in endometrial stromal cells, to verify the interaction of miR-183 with its target genes, and to confirm the role of miR-183 in the process of endometriosis. Using microarray analysis, we identified 27 differentially expressed genes (19 were upregulated and 8 downregulated), from which we selected 4 downregulated genes (ITGB1, AMIGO2, VAV3, and PSEN2) based on GO databases for functional analysis and significant pathway analysis. Western blotting analyses showed that integrin β1 (ITGB1), but not AMIGO2, was affected by miR-183 overexpression, whereas no protein expression of VAV3 and PSEN2 was detected. Luciferase reporter assay verified that ITGB1 is a target gene of miR-183. Moreover, we found that ITGB1 is overexpressed in the endometrium of endometriosis patients. Furthermore, overexpression of ITGB1 rescued the repressive effects of miR-183 on the invasiveness of endometrial stromal cells. These findings, together with the fact that ITGB1 is a critical factor for cell adhesion and invasiveness, suggest that miR-183 may be involved in the development of endometriosis by regulating ITGB1 in endometrial stromal cells.

No MeSH data available.


Related in: MedlinePlus

Overexpression of ITGB1 rescued the repressive effects of miR-183 on endometrial stromal cells, leading to elevated invasive abilities in transwell assays. Endometrial stromal cells were infected with miR-183-lentivirus, miR-183/ITGB1-lentivirus, and GFP-lentivirus in upper wells. After 24 hours, the number of cells that invaded through Matrigel was counted in at least 10 fields per well. (a) Representative photographs show that miR-183 inhibited the invasiveness of endometrial stromal cells, whereas ITGB1 partially rescued the repressive effects of miR-183. (b) Cell counting results indicate that ITGB1 overexpression rescued the repressive effects of miR-183. ∗P < 0.05 when compared to the negative control. Error bars represent ± SEM.
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fig5: Overexpression of ITGB1 rescued the repressive effects of miR-183 on endometrial stromal cells, leading to elevated invasive abilities in transwell assays. Endometrial stromal cells were infected with miR-183-lentivirus, miR-183/ITGB1-lentivirus, and GFP-lentivirus in upper wells. After 24 hours, the number of cells that invaded through Matrigel was counted in at least 10 fields per well. (a) Representative photographs show that miR-183 inhibited the invasiveness of endometrial stromal cells, whereas ITGB1 partially rescued the repressive effects of miR-183. (b) Cell counting results indicate that ITGB1 overexpression rescued the repressive effects of miR-183. ∗P < 0.05 when compared to the negative control. Error bars represent ± SEM.

Mentions: To ascertain that miR-183 regulates the function of endometrial stromal cells through its interaction with ITGB1, a rescue experiment was performed. Overexpression of ITGB1 partially rescued the repressive effects of miR-183, leading to elevated invasion abilities in the cells (Figure 5). This data indicated that miR-183 targets ITGB1, which in turn led to negative regulation on the invasive activity of endometrial stromal cells.


MiR-183 Regulates ITGB1P Expression and Promotes Invasion of Endometrial Stromal Cells.

Chen J, Gu L, Ni J, Hu P, Hu K, Shi YL - Biomed Res Int (2015)

Overexpression of ITGB1 rescued the repressive effects of miR-183 on endometrial stromal cells, leading to elevated invasive abilities in transwell assays. Endometrial stromal cells were infected with miR-183-lentivirus, miR-183/ITGB1-lentivirus, and GFP-lentivirus in upper wells. After 24 hours, the number of cells that invaded through Matrigel was counted in at least 10 fields per well. (a) Representative photographs show that miR-183 inhibited the invasiveness of endometrial stromal cells, whereas ITGB1 partially rescued the repressive effects of miR-183. (b) Cell counting results indicate that ITGB1 overexpression rescued the repressive effects of miR-183. ∗P < 0.05 when compared to the negative control. Error bars represent ± SEM.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4556833&req=5

fig5: Overexpression of ITGB1 rescued the repressive effects of miR-183 on endometrial stromal cells, leading to elevated invasive abilities in transwell assays. Endometrial stromal cells were infected with miR-183-lentivirus, miR-183/ITGB1-lentivirus, and GFP-lentivirus in upper wells. After 24 hours, the number of cells that invaded through Matrigel was counted in at least 10 fields per well. (a) Representative photographs show that miR-183 inhibited the invasiveness of endometrial stromal cells, whereas ITGB1 partially rescued the repressive effects of miR-183. (b) Cell counting results indicate that ITGB1 overexpression rescued the repressive effects of miR-183. ∗P < 0.05 when compared to the negative control. Error bars represent ± SEM.
Mentions: To ascertain that miR-183 regulates the function of endometrial stromal cells through its interaction with ITGB1, a rescue experiment was performed. Overexpression of ITGB1 partially rescued the repressive effects of miR-183, leading to elevated invasion abilities in the cells (Figure 5). This data indicated that miR-183 targets ITGB1, which in turn led to negative regulation on the invasive activity of endometrial stromal cells.

Bottom Line: We applied in the previous study miRNA microarray screening analysis to identify several differentially expressed miRNAs, including miR-183 in normal, eutopic, and ectopic endometrium.Western blotting analyses showed that integrin β1 (ITGB1), but not AMIGO2, was affected by miR-183 overexpression, whereas no protein expression of VAV3 and PSEN2 was detected.Moreover, we found that ITGB1 is overexpressed in the endometrium of endometriosis patients.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Reproductive Medicine, Department of Gynecology, Nanjing Maternity and Child Health Care Hospital Affiliated to Nanjing Medical University, 123 Tianfei Road, Nanjing, Jiangsu 210029, China.

ABSTRACT
We applied in the previous study miRNA microarray screening analysis to identify several differentially expressed miRNAs, including miR-183 in normal, eutopic, and ectopic endometrium. Knockdown of miR-183 expression induced the invasiveness and inhibition of apoptosis in endometrial stromal cells. The current study aims to identify the miR-183 targets with relevance to cell functions in endometrial stromal cells, to verify the interaction of miR-183 with its target genes, and to confirm the role of miR-183 in the process of endometriosis. Using microarray analysis, we identified 27 differentially expressed genes (19 were upregulated and 8 downregulated), from which we selected 4 downregulated genes (ITGB1, AMIGO2, VAV3, and PSEN2) based on GO databases for functional analysis and significant pathway analysis. Western blotting analyses showed that integrin β1 (ITGB1), but not AMIGO2, was affected by miR-183 overexpression, whereas no protein expression of VAV3 and PSEN2 was detected. Luciferase reporter assay verified that ITGB1 is a target gene of miR-183. Moreover, we found that ITGB1 is overexpressed in the endometrium of endometriosis patients. Furthermore, overexpression of ITGB1 rescued the repressive effects of miR-183 on the invasiveness of endometrial stromal cells. These findings, together with the fact that ITGB1 is a critical factor for cell adhesion and invasiveness, suggest that miR-183 may be involved in the development of endometriosis by regulating ITGB1 in endometrial stromal cells.

No MeSH data available.


Related in: MedlinePlus