Limits...
Cordycepin (3'-deoxyadenosine) suppressed HMGA2, Twist1 and ZEB1-dependent melanoma invasion and metastasis by targeting miR-33b.

Zhang P, Huang C, Fu C, Tian Y, Hu Y, Wang B, Strasner A, Song Y, Song E - Oncotarget (2015)

Bottom Line: Cordycepin, the active component of Cordyceps spp., has been identified to have anti-metastatic effect on tumor progression and thus possesses pharmacological and therapeutic potentials.Cordycepin-mediated miR-33b expression was dependent on LXR-RXR heterodimer activation. miR-33b directly binds to HMGA2, Twist1 and ZEB1 3'-UTR to suppress their expression.In spontaneous metastasis models, cordycepin suppressed tumor metastasis without altering primary tumor growth.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Luminescence and Real-Time Analytical Chemistry (Southwest University), Ministry of Education, College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, China.

ABSTRACT
Malignant melanoma, the most deadly form of skin cancer, has a high propensity for metastatic spread and is notoriously chemotherapy-resistant. Cordycepin, the active component of Cordyceps spp., has been identified to have anti-metastatic effect on tumor progression and thus possesses pharmacological and therapeutic potentials. However, the mechanisms of anti-metastatic effects of cordycepin at cellular levels remain elusive. We analyzed the effect of cordycepin on human melanoma miRNA expression profiles by miRNAarray and found that miR-33b was upregulated in highly-metastatic melanoma cell lines following cordycepin exposure. Cordycepin-mediated miR-33b expression was dependent on LXR-RXR heterodimer activation. miR-33b directly binds to HMGA2, Twist1 and ZEB1 3'-UTR to suppress their expression. The negative correlations between miR-33b levels and HMGA2, Twist1 or ZEB1 expression were detected in 72 patient melanoma tissue samples. By targeting HMGA2 and Twist1, miR-33b attenuated melanoma migration and invasiveness upon cordycepin exposure. miR-33b knockdown or ZEB1 overexpression reverted cordycepin-mediated mesenchymal-epithelial transition (MET), triggering the expression of N-cadherin. In spontaneous metastasis models, cordycepin suppressed tumor metastasis without altering primary tumor growth. We showed for the first time that targeting miRNA by cordycepin indicates a new mechanism of cordycepin-induced suppression of tumor metastasis and miR-33b/HMGA2/Twist1/ZEB1 axis plays critical roles in regulating melanoma dissemination.

No MeSH data available.


Related in: MedlinePlus

Cordycepin suppresses spontaneous metastasis through miR-33b(A) Bioluminescence images of primary tumor, lung, liver and bone from sh-NT or sh-miR-33b stable Lu1205 cells following 28 days post implantation. (B) Quantification of primary tumor weight (g) and metastasis to the lung, liver and bone as measured by bioluminescent intensity (represented by photons/sec). Cordycepin (3′-deoxyadenosine) was administrated i.p. daily to the nude mice in a dosage of 2 mg/kg body weight after the tumor inoculation. Ten mice were injected under each condition with each point representing a measurement from an individual mouse with the quartiles and median values indicated by a boxplot. P-values were calculated using a two-tailed Mann-Whitney U-test. (C) H&E (upper) staining of liver tissue and IHC (lower) staining of N-catenin in primary tumor. Arrow: metastatic tumor. (D) Prominent lung lesions of injected metastatic GFP-Lu1205 and GFP-A375 cells transfected with sh-NT and sh-miR-33b. Vesicle (ctrl) or cordycepin was administrated one week before tumor inoculation. Images were representative of six independent experiments. Number of lung lesions was quantified. **p < 0.01 compared with control. (E) Kaplan–Meier analysis comparing survival in sh-NT- and sh-miR-33b-transfected mice (left) and vector, HMGA2, Twist1 and ZEB1-transfected mice (right). Statistic significance between groups was analyzed by log-rank test. *p < 0.05 among groups. (F) Model of melanoma cancer metastasis suppressed via cordycepin-initiated signaling pathways.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4496401&req=5

Figure 7: Cordycepin suppresses spontaneous metastasis through miR-33b(A) Bioluminescence images of primary tumor, lung, liver and bone from sh-NT or sh-miR-33b stable Lu1205 cells following 28 days post implantation. (B) Quantification of primary tumor weight (g) and metastasis to the lung, liver and bone as measured by bioluminescent intensity (represented by photons/sec). Cordycepin (3′-deoxyadenosine) was administrated i.p. daily to the nude mice in a dosage of 2 mg/kg body weight after the tumor inoculation. Ten mice were injected under each condition with each point representing a measurement from an individual mouse with the quartiles and median values indicated by a boxplot. P-values were calculated using a two-tailed Mann-Whitney U-test. (C) H&E (upper) staining of liver tissue and IHC (lower) staining of N-catenin in primary tumor. Arrow: metastatic tumor. (D) Prominent lung lesions of injected metastatic GFP-Lu1205 and GFP-A375 cells transfected with sh-NT and sh-miR-33b. Vesicle (ctrl) or cordycepin was administrated one week before tumor inoculation. Images were representative of six independent experiments. Number of lung lesions was quantified. **p < 0.01 compared with control. (E) Kaplan–Meier analysis comparing survival in sh-NT- and sh-miR-33b-transfected mice (left) and vector, HMGA2, Twist1 and ZEB1-transfected mice (right). Statistic significance between groups was analyzed by log-rank test. *p < 0.05 among groups. (F) Model of melanoma cancer metastasis suppressed via cordycepin-initiated signaling pathways.

Mentions: To determine whether cordycepin influences spontaneous melanoma metastasis, we implanted nontargeting control (sh-NT) or sh-miR-33b-expressing Lu1205 cells to the right flank of nude mice and determined the primary tumor growth and metastasis to multiple organs by bioluminescence imaging. The 28-day tumor weight and size were comparable for control+sh-NT, cordycepin+sh-NT and cordycepin+sh-miR-33b groups (Figure 7A–7B). Nevertheless, cordycepin-treated melanoma metastasized less efficiently to lung, liver and bone. miR-33b knockdown in melanoma cells rescued the suppressive effect of cordycepin on metastasis (Figure 7A–7B). Cordycepin treatment or miR-33b knockdown did not affect the growth of the primary tumors, but did reduce the metastatic potential of cells in the spontaneous metastasis assay, therefore, indicating the involvement of miR-33b in the cordycepin-regulated melanoma invasiveness. To examine whether reduced metastasis of melanoma was due to the fact that primary tumor has underwent an epithelial differentiation, we stained primary tumor for N-cadherin. H&E staining verified bioluminescence results for the involvement of miR-33b in cordycepin-regulated melanoma liver metastasis (Figure 7C). N-cadherin expression was weak in primary tumor which was transfected by sh-NT and treated with cordycepin in comparison with vesicle-treated or cordycepin-treated/sh-miR-33b-transfected melanoma cells. miR-33b knockdown restored the robust N-cadherin staining in the cell membrane and cytosol of primary tumor in response to cordycepin treatment. This suggested that mesenchymal-epithelial transition (MET) was necessary for miR-33b-suppressed melanoma metastasis. Metastasis is a multistep process involving the early steps of EMT and intravasation followed by extravasation and colonization. To circumvent the early step of metastasis and specifically investigate whether miR-33b inhibits melanoma extravasation and colonization, we injected sh-NT or sh-miR-33b-expressing GFP-tagged melanoma cells via tail vein and monitored lung metastasis. In vesicle+sh-NT and cordycepin+sh-miR-33b cohorts, cells were completely cleared from the circulation in lung, 48-hr after tail vein injection. On the contrary, GFP-Lu1205 or GFP-A375 cells were trapped in the lung vein in cordycepin+sh-NT group, suggesting that cordycepin reduced cell extravasation by targeting miR-33b. After 20 days, appreciable amounts of tumor lesions formed in lung in control group (Figure 7D). Cordycepin treatment reduced the number of metastatic nodules in the lung of mice receiving Lu1205 and A375 cells by 78% and 80%, respectively. miR-33b knockdown rescued the abilities of Lu1205 and A375 cells to extravasate and colonize in the lung. Thus, miR-33b upregulated by cordycepin not only inhibited early stages of metastasis, but also repressed cell migration and invasion. The survival time of mice given cordycepin was significantly longer than that of control mice (Figure 7E). In contrast, miR-33b knockdown or HMGA2, Twist1 or ZEB1 overexpression significantly attenuated the effect of cordycepin on lifespan of tumor-bearing mice.


Cordycepin (3'-deoxyadenosine) suppressed HMGA2, Twist1 and ZEB1-dependent melanoma invasion and metastasis by targeting miR-33b.

Zhang P, Huang C, Fu C, Tian Y, Hu Y, Wang B, Strasner A, Song Y, Song E - Oncotarget (2015)

Cordycepin suppresses spontaneous metastasis through miR-33b(A) Bioluminescence images of primary tumor, lung, liver and bone from sh-NT or sh-miR-33b stable Lu1205 cells following 28 days post implantation. (B) Quantification of primary tumor weight (g) and metastasis to the lung, liver and bone as measured by bioluminescent intensity (represented by photons/sec). Cordycepin (3′-deoxyadenosine) was administrated i.p. daily to the nude mice in a dosage of 2 mg/kg body weight after the tumor inoculation. Ten mice were injected under each condition with each point representing a measurement from an individual mouse with the quartiles and median values indicated by a boxplot. P-values were calculated using a two-tailed Mann-Whitney U-test. (C) H&E (upper) staining of liver tissue and IHC (lower) staining of N-catenin in primary tumor. Arrow: metastatic tumor. (D) Prominent lung lesions of injected metastatic GFP-Lu1205 and GFP-A375 cells transfected with sh-NT and sh-miR-33b. Vesicle (ctrl) or cordycepin was administrated one week before tumor inoculation. Images were representative of six independent experiments. Number of lung lesions was quantified. **p < 0.01 compared with control. (E) Kaplan–Meier analysis comparing survival in sh-NT- and sh-miR-33b-transfected mice (left) and vector, HMGA2, Twist1 and ZEB1-transfected mice (right). Statistic significance between groups was analyzed by log-rank test. *p < 0.05 among groups. (F) Model of melanoma cancer metastasis suppressed via cordycepin-initiated signaling pathways.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4496401&req=5

Figure 7: Cordycepin suppresses spontaneous metastasis through miR-33b(A) Bioluminescence images of primary tumor, lung, liver and bone from sh-NT or sh-miR-33b stable Lu1205 cells following 28 days post implantation. (B) Quantification of primary tumor weight (g) and metastasis to the lung, liver and bone as measured by bioluminescent intensity (represented by photons/sec). Cordycepin (3′-deoxyadenosine) was administrated i.p. daily to the nude mice in a dosage of 2 mg/kg body weight after the tumor inoculation. Ten mice were injected under each condition with each point representing a measurement from an individual mouse with the quartiles and median values indicated by a boxplot. P-values were calculated using a two-tailed Mann-Whitney U-test. (C) H&E (upper) staining of liver tissue and IHC (lower) staining of N-catenin in primary tumor. Arrow: metastatic tumor. (D) Prominent lung lesions of injected metastatic GFP-Lu1205 and GFP-A375 cells transfected with sh-NT and sh-miR-33b. Vesicle (ctrl) or cordycepin was administrated one week before tumor inoculation. Images were representative of six independent experiments. Number of lung lesions was quantified. **p < 0.01 compared with control. (E) Kaplan–Meier analysis comparing survival in sh-NT- and sh-miR-33b-transfected mice (left) and vector, HMGA2, Twist1 and ZEB1-transfected mice (right). Statistic significance between groups was analyzed by log-rank test. *p < 0.05 among groups. (F) Model of melanoma cancer metastasis suppressed via cordycepin-initiated signaling pathways.
Mentions: To determine whether cordycepin influences spontaneous melanoma metastasis, we implanted nontargeting control (sh-NT) or sh-miR-33b-expressing Lu1205 cells to the right flank of nude mice and determined the primary tumor growth and metastasis to multiple organs by bioluminescence imaging. The 28-day tumor weight and size were comparable for control+sh-NT, cordycepin+sh-NT and cordycepin+sh-miR-33b groups (Figure 7A–7B). Nevertheless, cordycepin-treated melanoma metastasized less efficiently to lung, liver and bone. miR-33b knockdown in melanoma cells rescued the suppressive effect of cordycepin on metastasis (Figure 7A–7B). Cordycepin treatment or miR-33b knockdown did not affect the growth of the primary tumors, but did reduce the metastatic potential of cells in the spontaneous metastasis assay, therefore, indicating the involvement of miR-33b in the cordycepin-regulated melanoma invasiveness. To examine whether reduced metastasis of melanoma was due to the fact that primary tumor has underwent an epithelial differentiation, we stained primary tumor for N-cadherin. H&E staining verified bioluminescence results for the involvement of miR-33b in cordycepin-regulated melanoma liver metastasis (Figure 7C). N-cadherin expression was weak in primary tumor which was transfected by sh-NT and treated with cordycepin in comparison with vesicle-treated or cordycepin-treated/sh-miR-33b-transfected melanoma cells. miR-33b knockdown restored the robust N-cadherin staining in the cell membrane and cytosol of primary tumor in response to cordycepin treatment. This suggested that mesenchymal-epithelial transition (MET) was necessary for miR-33b-suppressed melanoma metastasis. Metastasis is a multistep process involving the early steps of EMT and intravasation followed by extravasation and colonization. To circumvent the early step of metastasis and specifically investigate whether miR-33b inhibits melanoma extravasation and colonization, we injected sh-NT or sh-miR-33b-expressing GFP-tagged melanoma cells via tail vein and monitored lung metastasis. In vesicle+sh-NT and cordycepin+sh-miR-33b cohorts, cells were completely cleared from the circulation in lung, 48-hr after tail vein injection. On the contrary, GFP-Lu1205 or GFP-A375 cells were trapped in the lung vein in cordycepin+sh-NT group, suggesting that cordycepin reduced cell extravasation by targeting miR-33b. After 20 days, appreciable amounts of tumor lesions formed in lung in control group (Figure 7D). Cordycepin treatment reduced the number of metastatic nodules in the lung of mice receiving Lu1205 and A375 cells by 78% and 80%, respectively. miR-33b knockdown rescued the abilities of Lu1205 and A375 cells to extravasate and colonize in the lung. Thus, miR-33b upregulated by cordycepin not only inhibited early stages of metastasis, but also repressed cell migration and invasion. The survival time of mice given cordycepin was significantly longer than that of control mice (Figure 7E). In contrast, miR-33b knockdown or HMGA2, Twist1 or ZEB1 overexpression significantly attenuated the effect of cordycepin on lifespan of tumor-bearing mice.

Bottom Line: Cordycepin, the active component of Cordyceps spp., has been identified to have anti-metastatic effect on tumor progression and thus possesses pharmacological and therapeutic potentials.Cordycepin-mediated miR-33b expression was dependent on LXR-RXR heterodimer activation. miR-33b directly binds to HMGA2, Twist1 and ZEB1 3'-UTR to suppress their expression.In spontaneous metastasis models, cordycepin suppressed tumor metastasis without altering primary tumor growth.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Luminescence and Real-Time Analytical Chemistry (Southwest University), Ministry of Education, College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, China.

ABSTRACT
Malignant melanoma, the most deadly form of skin cancer, has a high propensity for metastatic spread and is notoriously chemotherapy-resistant. Cordycepin, the active component of Cordyceps spp., has been identified to have anti-metastatic effect on tumor progression and thus possesses pharmacological and therapeutic potentials. However, the mechanisms of anti-metastatic effects of cordycepin at cellular levels remain elusive. We analyzed the effect of cordycepin on human melanoma miRNA expression profiles by miRNAarray and found that miR-33b was upregulated in highly-metastatic melanoma cell lines following cordycepin exposure. Cordycepin-mediated miR-33b expression was dependent on LXR-RXR heterodimer activation. miR-33b directly binds to HMGA2, Twist1 and ZEB1 3'-UTR to suppress their expression. The negative correlations between miR-33b levels and HMGA2, Twist1 or ZEB1 expression were detected in 72 patient melanoma tissue samples. By targeting HMGA2 and Twist1, miR-33b attenuated melanoma migration and invasiveness upon cordycepin exposure. miR-33b knockdown or ZEB1 overexpression reverted cordycepin-mediated mesenchymal-epithelial transition (MET), triggering the expression of N-cadherin. In spontaneous metastasis models, cordycepin suppressed tumor metastasis without altering primary tumor growth. We showed for the first time that targeting miRNA by cordycepin indicates a new mechanism of cordycepin-induced suppression of tumor metastasis and miR-33b/HMGA2/Twist1/ZEB1 axis plays critical roles in regulating melanoma dissemination.

No MeSH data available.


Related in: MedlinePlus