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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

Confirmation of microarray data with Western blotting analyses. The endometrial stromal cells transfected with miR-183-lentivirus (low dose or high dose), In-miR-183-lentivirus (low dose or high dose), and GFP-lentivirus (NC, normal control) were used to examine the expression alterations of integrin β1 (a) and AMIGO2 (b). β-actin served as protein loading control. Compared with the control group, integrin β1 expression in the miR-183-overexpression group was significantly decreased, whereas it increased sharply in cells with low miR-183 expression. AMIGO2 was largely not regulated by miR-183. ∗P < 0.05 when compared to the negative control. Error bars represent ± SEM.
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fig2: Confirmation of microarray data with Western blotting analyses. The endometrial stromal cells transfected with miR-183-lentivirus (low dose or high dose), In-miR-183-lentivirus (low dose or high dose), and GFP-lentivirus (NC, normal control) were used to examine the expression alterations of integrin β1 (a) and AMIGO2 (b). β-actin served as protein loading control. Compared with the control group, integrin β1 expression in the miR-183-overexpression group was significantly decreased, whereas it increased sharply in cells with low miR-183 expression. AMIGO2 was largely not regulated by miR-183. ∗P < 0.05 when compared to the negative control. Error bars represent ± SEM.

Mentions: Western blotting (Figure 2) was used to confirm their alterations on protein levels. Using the initial sample sets and the criteria of ≥1.5-fold of change, integrin β1 yielded consistent results by the two technologies. Compared to the control group, the levels of integrin β1 showed a significant decrease in the miR-183-overexpression group, whereas the miR-183-downexpressing group (P < 0.05) showed a sharp increase. Although endometrial stromal cells express AMIGO2 protein, the amount was largely unchanged in the miR-183 up- or downexpression groups, suggesting it was not regulated by miR-183. On the other hand, no protein expression of Presenilin 2 and VAV3 was detected in endometrial stromal cells (data not shown) and these two genes were not further studied.


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)

Confirmation of microarray data with Western blotting analyses. The endometrial stromal cells transfected with miR-183-lentivirus (low dose or high dose), In-miR-183-lentivirus (low dose or high dose), and GFP-lentivirus (NC, normal control) were used to examine the expression alterations of integrin β1 (a) and AMIGO2 (b). β-actin served as protein loading control. Compared with the control group, integrin β1 expression in the miR-183-overexpression group was significantly decreased, whereas it increased sharply in cells with low miR-183 expression. AMIGO2 was largely not regulated by 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

fig2: Confirmation of microarray data with Western blotting analyses. The endometrial stromal cells transfected with miR-183-lentivirus (low dose or high dose), In-miR-183-lentivirus (low dose or high dose), and GFP-lentivirus (NC, normal control) were used to examine the expression alterations of integrin β1 (a) and AMIGO2 (b). β-actin served as protein loading control. Compared with the control group, integrin β1 expression in the miR-183-overexpression group was significantly decreased, whereas it increased sharply in cells with low miR-183 expression. AMIGO2 was largely not regulated by miR-183. ∗P < 0.05 when compared to the negative control. Error bars represent ± SEM.
Mentions: Western blotting (Figure 2) was used to confirm their alterations on protein levels. Using the initial sample sets and the criteria of ≥1.5-fold of change, integrin β1 yielded consistent results by the two technologies. Compared to the control group, the levels of integrin β1 showed a significant decrease in the miR-183-overexpression group, whereas the miR-183-downexpressing group (P < 0.05) showed a sharp increase. Although endometrial stromal cells express AMIGO2 protein, the amount was largely unchanged in the miR-183 up- or downexpression groups, suggesting it was not regulated by miR-183. On the other hand, no protein expression of Presenilin 2 and VAV3 was detected in endometrial stromal cells (data not shown) and these two genes were not further studied.

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