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MiR-203 down-regulates Rap1A and suppresses cell proliferation, adhesion and invasion in prostate cancer.

Xiang J, Bian C, Wang H, Huang S, Wu D - J. Exp. Clin. Cancer Res. (2015)

Bottom Line: Here, we confirmed that the expression of miR-203 was significantly downregulated in prostate cancer specimens compared with matched adjacent normal prostate specimens.In addition, we identified Rap1A as a direct target suppressed by miR-203, and there was an inverse relationship between the expression of miR-203 and Rap1A in PCa.These findings provide further evidence that a crucial role for miR-203 in inhibiting metastasis of PCa through the suppression of Rap1A expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, Tongji Hospital, Tongji University School of Medicine, NO 389 Xinchun road, Shanghai, 200065, China. zhuxufifa2007@163.com.

ABSTRACT

Objective: Evidence supports an important role for miR-203 in the regulation of the proliferation, migration and invasion of prostate cancer (PCa) cells. However, the exact mechanisms of miR-203 in PCa are not entirely clear.

Methods: We examined the expression of miR-203 in prostate cancer tissues, adjacent normal tissues, PCa cell lines and normal prostate epithelial cells by qRT-PCR. Then, the effects of miR-203 or Rap1A on proliferation, adhesion and invasion of PCa cells were assayed using CKK-8, adhesion analysis, and transwell invasion assays. Luciferase reporter assay was performed to assess miR-203 binding to Rap1A mRNA. Tumor growth was assessed by subcutaneous inoculation of cells into BALB/c nude mice.

Results: Here, we confirmed that the expression of miR-203 was significantly downregulated in prostate cancer specimens compared with matched adjacent normal prostate specimens. Mechanistic dissection revealed that miR-203 mediated cell proliferation, adhesion and invasion in vitro, and tumor growth in vivo, as evidenced by reduced RAC1, p-PAK1, and p-MEK1 expression. In addition, we identified Rap1A as a direct target suppressed by miR-203, and there was an inverse relationship between the expression of miR-203 and Rap1A in PCa. Knockdown of Rap1A phenocopied the effects of miR-203 on PCa cell growth and invasion. Furthermore, Rap1A over-expression in PCa cells partially reversed the effects of miR-203-expression on cell adhesion and invasion.

Conclusions: These findings provide further evidence that a crucial role for miR-203 in inhibiting metastasis of PCa through the suppression of Rap1A expression.

No MeSH data available.


Related in: MedlinePlus

MiR-203 inhibits tumor growth in vivo. (A) Representative anatomical photos of xenograft tumors in nude mice injected subcutaneously with DU145 cells infected with vector or miR-203. Tumor weight was measured at the same time. (B) Rap1A protein expression in xenografts from vector control groups and miR-203 over-expression groups was assessed by western blot analysis. Lanes 1, 3, 5, 7 were samples from the vector control group. Lanes 2, 4, 6, 8 were samples from the miR-203 over-expression group. ** p < 0.01.
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Fig6: MiR-203 inhibits tumor growth in vivo. (A) Representative anatomical photos of xenograft tumors in nude mice injected subcutaneously with DU145 cells infected with vector or miR-203. Tumor weight was measured at the same time. (B) Rap1A protein expression in xenografts from vector control groups and miR-203 over-expression groups was assessed by western blot analysis. Lanes 1, 3, 5, 7 were samples from the vector control group. Lanes 2, 4, 6, 8 were samples from the miR-203 over-expression group. ** p < 0.01.

Mentions: To investigate the contribution of miR-203 in vivo, we injected DU 145cells with miR-203 or control vector subcutaneously into the flanks of nude mice. In this assay, each experimental mouse bearing miR-203-overexpressing and control vector DU 145 cells, respectively, on the right or left dorsal thigh began to exhibit differences in tumor growth between the two sides within two weeks, and the difference continued to increase through the endpoint, at which tumors of the miR-203-overexpressing group displayed a smaller tumor size and weight (Figure 6A). To further explore the in vivo relevance of these observations, we assessed the expression of Rap1A proteins in homogenates from xenograft tumors. Rap1A protein was reduced in DU 145-miR-203 cells compared with control cells (Figure 6B). Taken together, these findings were consistent with the in vitro results and indicated that miR-203 has the ability to suppress PCa cell growth in vivo.Figure 6


MiR-203 down-regulates Rap1A and suppresses cell proliferation, adhesion and invasion in prostate cancer.

Xiang J, Bian C, Wang H, Huang S, Wu D - J. Exp. Clin. Cancer Res. (2015)

MiR-203 inhibits tumor growth in vivo. (A) Representative anatomical photos of xenograft tumors in nude mice injected subcutaneously with DU145 cells infected with vector or miR-203. Tumor weight was measured at the same time. (B) Rap1A protein expression in xenografts from vector control groups and miR-203 over-expression groups was assessed by western blot analysis. Lanes 1, 3, 5, 7 were samples from the vector control group. Lanes 2, 4, 6, 8 were samples from the miR-203 over-expression group. ** p < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4321708&req=5

Fig6: MiR-203 inhibits tumor growth in vivo. (A) Representative anatomical photos of xenograft tumors in nude mice injected subcutaneously with DU145 cells infected with vector or miR-203. Tumor weight was measured at the same time. (B) Rap1A protein expression in xenografts from vector control groups and miR-203 over-expression groups was assessed by western blot analysis. Lanes 1, 3, 5, 7 were samples from the vector control group. Lanes 2, 4, 6, 8 were samples from the miR-203 over-expression group. ** p < 0.01.
Mentions: To investigate the contribution of miR-203 in vivo, we injected DU 145cells with miR-203 or control vector subcutaneously into the flanks of nude mice. In this assay, each experimental mouse bearing miR-203-overexpressing and control vector DU 145 cells, respectively, on the right or left dorsal thigh began to exhibit differences in tumor growth between the two sides within two weeks, and the difference continued to increase through the endpoint, at which tumors of the miR-203-overexpressing group displayed a smaller tumor size and weight (Figure 6A). To further explore the in vivo relevance of these observations, we assessed the expression of Rap1A proteins in homogenates from xenograft tumors. Rap1A protein was reduced in DU 145-miR-203 cells compared with control cells (Figure 6B). Taken together, these findings were consistent with the in vitro results and indicated that miR-203 has the ability to suppress PCa cell growth in vivo.Figure 6

Bottom Line: Here, we confirmed that the expression of miR-203 was significantly downregulated in prostate cancer specimens compared with matched adjacent normal prostate specimens.In addition, we identified Rap1A as a direct target suppressed by miR-203, and there was an inverse relationship between the expression of miR-203 and Rap1A in PCa.These findings provide further evidence that a crucial role for miR-203 in inhibiting metastasis of PCa through the suppression of Rap1A expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, Tongji Hospital, Tongji University School of Medicine, NO 389 Xinchun road, Shanghai, 200065, China. zhuxufifa2007@163.com.

ABSTRACT

Objective: Evidence supports an important role for miR-203 in the regulation of the proliferation, migration and invasion of prostate cancer (PCa) cells. However, the exact mechanisms of miR-203 in PCa are not entirely clear.

Methods: We examined the expression of miR-203 in prostate cancer tissues, adjacent normal tissues, PCa cell lines and normal prostate epithelial cells by qRT-PCR. Then, the effects of miR-203 or Rap1A on proliferation, adhesion and invasion of PCa cells were assayed using CKK-8, adhesion analysis, and transwell invasion assays. Luciferase reporter assay was performed to assess miR-203 binding to Rap1A mRNA. Tumor growth was assessed by subcutaneous inoculation of cells into BALB/c nude mice.

Results: Here, we confirmed that the expression of miR-203 was significantly downregulated in prostate cancer specimens compared with matched adjacent normal prostate specimens. Mechanistic dissection revealed that miR-203 mediated cell proliferation, adhesion and invasion in vitro, and tumor growth in vivo, as evidenced by reduced RAC1, p-PAK1, and p-MEK1 expression. In addition, we identified Rap1A as a direct target suppressed by miR-203, and there was an inverse relationship between the expression of miR-203 and Rap1A in PCa. Knockdown of Rap1A phenocopied the effects of miR-203 on PCa cell growth and invasion. Furthermore, Rap1A over-expression in PCa cells partially reversed the effects of miR-203-expression on cell adhesion and invasion.

Conclusions: These findings provide further evidence that a crucial role for miR-203 in inhibiting metastasis of PCa through the suppression of Rap1A expression.

No MeSH data available.


Related in: MedlinePlus