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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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The down-regulation of miR-143 repressed PC-3-M metastasis in vivo.A, The down-regulation of miR-143 in PC-3-M stable cell lines on days 0 and 60 by shRNA was verified by qRT-PCR. B, Systemic metastases of PC-3-M cells was measured in mice by bioluminescence imaging 30 days post-implant. C, The liver metastasis of PC-3-M cells was measured by macroscopic and bioluminescent methods after autopsy. D, Histologic analysis of metastatic lesions in the ribs of nude mice was carried by H&E-staining in which tumors were marked as “T”, bone was “B” and muscle was “M”.
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Figure 4: The down-regulation of miR-143 repressed PC-3-M metastasis in vivo.A, The down-regulation of miR-143 in PC-3-M stable cell lines on days 0 and 60 by shRNA was verified by qRT-PCR. B, Systemic metastases of PC-3-M cells was measured in mice by bioluminescence imaging 30 days post-implant. C, The liver metastasis of PC-3-M cells was measured by macroscopic and bioluminescent methods after autopsy. D, Histologic analysis of metastatic lesions in the ribs of nude mice was carried by H&E-staining in which tumors were marked as “T”, bone was “B” and muscle was “M”.

Mentions: To further explore the effect of miR-143 on prostate cancer metastasis in vivo, PC-3-M cells were stably transfected with NC or miR-143 inhibitor and subcutaneously injected into nude mice. As shown in Figure 4A, the relative expression of miR-143 in PC-3-M cells transfected with the miR-143 inhibitor was stably about 10-fold lower compared with cells transfected with NC at least 60 days. The development and metastasis of tumors in vivo were monitored by visualizing the bioluminescence emitted from the luciferase-tagged tumors on day 30. Mice injected with the miR-143 inhibitor PC-3-M cells developed fewer systemic metastasis (2/10 versus 8/10), especially fewer macroscopic and bioluminescent nodes in the liver (3/10 versus 9/10), compared with mice implanted with NC cells (Figure 4B,C, Table 3). Histological confirmations were made by H&E-stainning (Figure 4D). Mice injected with the miR-143 NC PC-3-M cells developed severe metastatic lesions in the ribs of nude mice, and only a litter bone and muscle were residual. However, no bone metastasis was seen in the mice implanted with miR-143 inhibitor cells. These findings suggested that the down-regulation of miR-143 inhibited prostate cancer cells metastasis in vivo.


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)

The down-regulation of miR-143 repressed PC-3-M metastasis in vivo.A, The down-regulation of miR-143 in PC-3-M stable cell lines on days 0 and 60 by shRNA was verified by qRT-PCR. B, Systemic metastases of PC-3-M cells was measured in mice by bioluminescence imaging 30 days post-implant. C, The liver metastasis of PC-3-M cells was measured by macroscopic and bioluminescent methods after autopsy. D, Histologic analysis of metastatic lesions in the ribs of nude mice was carried by H&E-staining in which tumors were marked as “T”, bone was “B” and muscle was “M”.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3585861&req=5

Figure 4: The down-regulation of miR-143 repressed PC-3-M metastasis in vivo.A, The down-regulation of miR-143 in PC-3-M stable cell lines on days 0 and 60 by shRNA was verified by qRT-PCR. B, Systemic metastases of PC-3-M cells was measured in mice by bioluminescence imaging 30 days post-implant. C, The liver metastasis of PC-3-M cells was measured by macroscopic and bioluminescent methods after autopsy. D, Histologic analysis of metastatic lesions in the ribs of nude mice was carried by H&E-staining in which tumors were marked as “T”, bone was “B” and muscle was “M”.
Mentions: To further explore the effect of miR-143 on prostate cancer metastasis in vivo, PC-3-M cells were stably transfected with NC or miR-143 inhibitor and subcutaneously injected into nude mice. As shown in Figure 4A, the relative expression of miR-143 in PC-3-M cells transfected with the miR-143 inhibitor was stably about 10-fold lower compared with cells transfected with NC at least 60 days. The development and metastasis of tumors in vivo were monitored by visualizing the bioluminescence emitted from the luciferase-tagged tumors on day 30. Mice injected with the miR-143 inhibitor PC-3-M cells developed fewer systemic metastasis (2/10 versus 8/10), especially fewer macroscopic and bioluminescent nodes in the liver (3/10 versus 9/10), compared with mice implanted with NC cells (Figure 4B,C, Table 3). Histological confirmations were made by H&E-stainning (Figure 4D). Mice injected with the miR-143 NC PC-3-M cells developed severe metastatic lesions in the ribs of nude mice, and only a litter bone and muscle were residual. However, no bone metastasis was seen in the mice implanted with miR-143 inhibitor cells. These findings suggested that the down-regulation of miR-143 inhibited prostate cancer cells metastasis in vivo.

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