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Up-regulated microRNA-143 in cancer stem cells differentiation promotes prostate cancer cells metastasis by modulating FNDC3B expression.

Fan X, Chen X, Deng W, Zhong G, Cai Q, Lin T - BMC Cancer (2013)

Bottom Line: The expression of miR-143 and the migration capability were reduced in PC-3 sphere cells and progressively increased during sphere re-adherent culture.Fibronectin type III domain containing 3B (FNDC3B), which regulates cell motility, was identified as a target of miR-143.These data demonstrate for the first time that miR-143 was up-regulated during the differentiation of prostate cancer stem cells and promoted prostate cancer metastasis by repressing FNDC3B expression.

View Article: PubMed Central - HTML - PubMed

Affiliation: Center of medical research, Sun Yat-Sen memorial hospital, Sun Yat-Sen University, Guangzhou, 510120, China.

ABSTRACT

Background: Metastatic prostate cancer is a leading cause of cancer-related death in men. Cancer stem cells (CSCs) are involved in tumor progression and metastasis, including in prostate cancer. There is an obvious and urgent need for effective cancer stem cells specific therapies in metastatic prostate cancer. MicroRNAs (miRNAs) are an important class of pervasive genes that are involved in a variety of biological functions, especially in cancer. The goal of this study was to identify miRNAs involved in prostate cancer metastasis and cancer stem cells.

Methods: A microarray and qRT-PCR were performed to investigate the miRNA expression profiles in PC-3 sphere cells and adherent cells. A transwell assay was used to evaluate the migration of PC-3 sphere cells and adherent cells. MiR-143 was silenced with antisense oligonucleotides in PC-3, PC-3-M and LNCaP cells. The role of miR-143 in prostate cancer metastasis was measured by wound-healing and transwell assays in vitro and bioluminescence imaging in vivo. Bioinformatics and luciferase report assays were used to identify the target of miR-143.

Results: The expression of miR-143 and the migration capability were reduced in PC-3 sphere cells and progressively increased during sphere re-adherent culture. Moreover, the down-regulation of miR-143 suppressed prostate cancer cells migration and invasion in vitro and systemically inhibited metastasis in vivo. Fibronectin type III domain containing 3B (FNDC3B), which regulates cell motility, was identified as a target of miR-143. The inhibition of miR-143 increased the expression of FNDC3B protein but not FNDC3B mRNA in vitro and vivo.

Conclusions: These data demonstrate for the first time that miR-143 was up-regulated during the differentiation of prostate cancer stem cells and promoted prostate cancer metastasis by repressing FNDC3B expression. This sheds a new insight into the post-transcriptional regulation of cancer stem cells differentiation by miRNAs, a potential approach for the treatment of prostate cancer.

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MiR-143 targeted oncogene FNDC3B.A, The putative miR-143-binding sequence was in the 3′-UTR of FNDC3B mRNA. B, The dual luciferase report assay of HEK-293 T cells cotransfected with psi-CHECK2-FNDC3B and miRNA (miR-143, NC). The Renilla luciferase signals were normalized to the internal firefly luciferase transfection control. C, The mRNA and protein expression of FNDC3B in PC-3-M cells transfected with miR-143 inhibitor or NC was analyzed by RT-PCR (above) and western blotting (below). D, The expression of miR-143 and FNDC3B in the tumor tissues of nude mice were analyzed by RT-PCR (above) and western blotting (below). U6 and GAPDH were internal control. (* P < 0.05).
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Figure 5: MiR-143 targeted oncogene FNDC3B.A, The putative miR-143-binding sequence was in the 3′-UTR of FNDC3B mRNA. B, The dual luciferase report assay of HEK-293 T cells cotransfected with psi-CHECK2-FNDC3B and miRNA (miR-143, NC). The Renilla luciferase signals were normalized to the internal firefly luciferase transfection control. C, The mRNA and protein expression of FNDC3B in PC-3-M cells transfected with miR-143 inhibitor or NC was analyzed by RT-PCR (above) and western blotting (below). D, The expression of miR-143 and FNDC3B in the tumor tissues of nude mice were analyzed by RT-PCR (above) and western blotting (below). U6 and GAPDH were internal control. (* P < 0.05).

Mentions: To elucidate the mechanisms through which miR-143 induced prostate cancer cells metastasis, two publicly available algorithms were used to help predict miR-143 targets. Among the approximately 200 candidate genes, fibronectin type III domain containing 3B (FNDC3B) was a high-scoring candidate. FNDC3B is a member of the fibronectin family, which regulates cell motility, and is down-regulated in tumor cells with high metastatic potential [19]. Zhang et al. [20] reported that up-regulated miRNA-143 enhanced hepatocarcinoma metastasis by repressing FNDC3B expression. Thus, we focused on the possible regulation of FNDC3B by miR-143. As shown in Figure 5A, miR-143 was partially complementary to the FNDC3B mRNA 3′ untranslated region (UTR) element. Consequently, a luciferase reporter assay was performed to verify whether miR-143 could directly target the FNDC3B 3′-UTR. The luciferase activity of psi-CHECK2-FNDC3B was obviously decreased in the presence of miR-143, whereas no significant reduction was observed when cells were cotransfected with NC (Figure 5B). Moreover, we evaluated the effects of miR-143 on FNDC3B mRNA and protein levels in PC-3-M cells by RT-PCR and western blotting. FNDC3B mRNA levels showed no significant change whereas protein levels were increased in PC-3-M cells transfected with miR-143-inhibitor (Figure 5C). We also investigate the expression of miR-143 and FNDC3B in the tumor tissue of nude mice by RT-PCR and western blotting. As expected, the expression of miR-143 was significantly decreased whereas FNDC3B protein levels were enhanced in the tumor tissue of the group of miR-143-inhibitor (Figure 5D). Taken together, these findings suggested that miR-143 might modulate prostate cancer metastasis by targeting FNDC3B.


Up-regulated microRNA-143 in cancer stem cells differentiation promotes prostate cancer cells metastasis by modulating FNDC3B expression.

Fan X, Chen X, Deng W, Zhong G, Cai Q, Lin T - BMC Cancer (2013)

MiR-143 targeted oncogene FNDC3B.A, The putative miR-143-binding sequence was in the 3′-UTR of FNDC3B mRNA. B, The dual luciferase report assay of HEK-293 T cells cotransfected with psi-CHECK2-FNDC3B and miRNA (miR-143, NC). The Renilla luciferase signals were normalized to the internal firefly luciferase transfection control. C, The mRNA and protein expression of FNDC3B in PC-3-M cells transfected with miR-143 inhibitor or NC was analyzed by RT-PCR (above) and western blotting (below). D, The expression of miR-143 and FNDC3B in the tumor tissues of nude mice were analyzed by RT-PCR (above) and western blotting (below). U6 and GAPDH were internal control. (* P < 0.05).
© Copyright Policy - open-access
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Figure 5: MiR-143 targeted oncogene FNDC3B.A, The putative miR-143-binding sequence was in the 3′-UTR of FNDC3B mRNA. B, The dual luciferase report assay of HEK-293 T cells cotransfected with psi-CHECK2-FNDC3B and miRNA (miR-143, NC). The Renilla luciferase signals were normalized to the internal firefly luciferase transfection control. C, The mRNA and protein expression of FNDC3B in PC-3-M cells transfected with miR-143 inhibitor or NC was analyzed by RT-PCR (above) and western blotting (below). D, The expression of miR-143 and FNDC3B in the tumor tissues of nude mice were analyzed by RT-PCR (above) and western blotting (below). U6 and GAPDH were internal control. (* P < 0.05).
Mentions: To elucidate the mechanisms through which miR-143 induced prostate cancer cells metastasis, two publicly available algorithms were used to help predict miR-143 targets. Among the approximately 200 candidate genes, fibronectin type III domain containing 3B (FNDC3B) was a high-scoring candidate. FNDC3B is a member of the fibronectin family, which regulates cell motility, and is down-regulated in tumor cells with high metastatic potential [19]. Zhang et al. [20] reported that up-regulated miRNA-143 enhanced hepatocarcinoma metastasis by repressing FNDC3B expression. Thus, we focused on the possible regulation of FNDC3B by miR-143. As shown in Figure 5A, miR-143 was partially complementary to the FNDC3B mRNA 3′ untranslated region (UTR) element. Consequently, a luciferase reporter assay was performed to verify whether miR-143 could directly target the FNDC3B 3′-UTR. The luciferase activity of psi-CHECK2-FNDC3B was obviously decreased in the presence of miR-143, whereas no significant reduction was observed when cells were cotransfected with NC (Figure 5B). Moreover, we evaluated the effects of miR-143 on FNDC3B mRNA and protein levels in PC-3-M cells by RT-PCR and western blotting. FNDC3B mRNA levels showed no significant change whereas protein levels were increased in PC-3-M cells transfected with miR-143-inhibitor (Figure 5C). We also investigate the expression of miR-143 and FNDC3B in the tumor tissue of nude mice by RT-PCR and western blotting. As expected, the expression of miR-143 was significantly decreased whereas FNDC3B protein levels were enhanced in the tumor tissue of the group of miR-143-inhibitor (Figure 5D). Taken together, these findings suggested that miR-143 might modulate prostate cancer metastasis by targeting FNDC3B.

Bottom Line: The expression of miR-143 and the migration capability were reduced in PC-3 sphere cells and progressively increased during sphere re-adherent culture.Fibronectin type III domain containing 3B (FNDC3B), which regulates cell motility, was identified as a target of miR-143.These data demonstrate for the first time that miR-143 was up-regulated during the differentiation of prostate cancer stem cells and promoted prostate cancer metastasis by repressing FNDC3B expression.

View Article: PubMed Central - HTML - PubMed

Affiliation: Center of medical research, Sun Yat-Sen memorial hospital, Sun Yat-Sen University, Guangzhou, 510120, China.

ABSTRACT

Background: Metastatic prostate cancer is a leading cause of cancer-related death in men. Cancer stem cells (CSCs) are involved in tumor progression and metastasis, including in prostate cancer. There is an obvious and urgent need for effective cancer stem cells specific therapies in metastatic prostate cancer. MicroRNAs (miRNAs) are an important class of pervasive genes that are involved in a variety of biological functions, especially in cancer. The goal of this study was to identify miRNAs involved in prostate cancer metastasis and cancer stem cells.

Methods: A microarray and qRT-PCR were performed to investigate the miRNA expression profiles in PC-3 sphere cells and adherent cells. A transwell assay was used to evaluate the migration of PC-3 sphere cells and adherent cells. MiR-143 was silenced with antisense oligonucleotides in PC-3, PC-3-M and LNCaP cells. The role of miR-143 in prostate cancer metastasis was measured by wound-healing and transwell assays in vitro and bioluminescence imaging in vivo. Bioinformatics and luciferase report assays were used to identify the target of miR-143.

Results: The expression of miR-143 and the migration capability were reduced in PC-3 sphere cells and progressively increased during sphere re-adherent culture. Moreover, the down-regulation of miR-143 suppressed prostate cancer cells migration and invasion in vitro and systemically inhibited metastasis in vivo. Fibronectin type III domain containing 3B (FNDC3B), which regulates cell motility, was identified as a target of miR-143. The inhibition of miR-143 increased the expression of FNDC3B protein but not FNDC3B mRNA in vitro and vivo.

Conclusions: These data demonstrate for the first time that miR-143 was up-regulated during the differentiation of prostate cancer stem cells and promoted prostate cancer metastasis by repressing FNDC3B expression. This sheds a new insight into the post-transcriptional regulation of cancer stem cells differentiation by miRNAs, a potential approach for the treatment of prostate cancer.

Show MeSH
Related in: MedlinePlus