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Cucurbitacin I blocks cerebrospinal fluid and platelet derived growth factor-BB stimulation of leptomeningeal and meningioma DNA synthesis.

Johnson MD, O'Connell MJ, Walter K - BMC Complement Altern Med (2013)

Bottom Line: Co-administration of cucurbitacin I (250 nM) produces a significant blockade of this effect.Cucurbitacin I alone also produced a significant reduction in basal DNA synthesis.Cucurbitacin I had no effect on basal phosphorylation of STAT3 but reduced phorphorylated p44/42 MAPK in 2 grade II meningioma cells lines.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Neuropathology, Department of Pathology and Laboratory Medicine, University of Rochester School of Medicine and Dentistry, Box 626, 601 Elmwood Ave,, Rochester, NY 14642, USA. mahlon_johnson@urmc.rochester.edu.

ABSTRACT

Background: Currently, there are no consistently effective chemotherapies for recurrent and inoperable meningiomas. Recently, cucurbitacin I (JSI-124), a naturally occurring tetracyclic triterpenoid compound used as folk medicines has been found to have cytoxic and anti-proliferative properties in several malignancies thru inhibition of activator of transcription (STAT3) activation. Previously, we have found STAT3 to be activated in meningiomas, particularly higher grade tumors.

Methods: Primary leptomeningeal cultures were established from 17, 20 and 22 week human fetuses and meningioma cell cultures were established from 6 World Health Organization (WHO) grade I or II meningiomas. Cells were treated with cerebrospinal fluid from patients without neurologic disease. The effects of cucurbitacin I on cerebrospinal fluid stimulation of meningioma cell DNA synthesis phosphorylation/activation of JAK1, STAT3, pMEK1/2, p44/42MAPK, Akt, mTOR, Rb and caspase 3 activation were analyzed in human leptomeningeal and meningioma cells.

Results: Cerebrospinal fluid significantly stimulated DNA synthesis in leptomeningeal cells. Co-administration of cucurbitacin I (250 nM) produces a significant blockade of this effect. Cucurbitacin I alone also produced a significant reduction in basal DNA synthesis. In grade I and II meningiomas, cerebrospinal fluid also significantly stimulated DNA synthesis. Co-administration of cucurbitacin I (250 nM) blocked this effect.In the leptomeningeal cultures, cerebrospinal fluid stimulated STAT3 phosphorylation but not p44/42MAPK, Akt or mTOR. Cucurbitacin I had no effect on basal STAT3 phosphorylation but co-administration with cerebrospinal fluid blocked cerebrospinal fluid stimulation of STAT3 phosphorylation in each. In the grade I meningiomas, cerebrospinal fluid stimulated phosphorylation of STAT3 and decreased MEK1/2 and cucurbitacin I had no effect on basal STAT3, p44/42MAPK, Akt, JAK1, mTOR, or Rb phosphorylation. In the grade II meningiomas, cerebrospinal fluid stimulated STAT3 phosphorylation in all and reduced phosphorylation of MEK1/2 in all and p44/42MAPK in one. Cucurbitacin I had no effect on basal phosphorylation of STAT3 but reduced phorphorylated p44/42 MAPK in 2 grade II meningioma cells lines.

Conclusions: These studies raise the possibility that cucurbitacin I might have value as an adjunct chemotherapy. Additional studies are warranted to evaluate the effects of cucurbitacin I on meningiomas in vivo.

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Effects of cucurbitacin I on cerebrospinal fluid stimulation of leptomeningeal and meningioma cell DNA synthesis. a. Cells from 17 (L1), 20 (L2) and 22 (L3) wk fetal leptomeninges (L), were treated with CSF without or with cucurbitacin I (250 nM) for 72 hrs. CyQUANT analysis of DNA measured with a fluorescence microplate reader with filters appropriate for ~480 nm excitation and ~520 nm emission maxima. Changes in DNA fluorescence correlate with cell proliferation. C = control, CSF = cerebrospinal fluid. * P < 0.05, *** P < 0.005, **** P < 0.001, ***** P < 0.0005. b. Cells from WHO grade I (M1-M3) and WHO grade II (M4-M5) meningiomas were treated as above. * P < 0.05, *** P < 0.005, **** P < 0.001, ***** P < 0.0005.
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Figure 2: Effects of cucurbitacin I on cerebrospinal fluid stimulation of leptomeningeal and meningioma cell DNA synthesis. a. Cells from 17 (L1), 20 (L2) and 22 (L3) wk fetal leptomeninges (L), were treated with CSF without or with cucurbitacin I (250 nM) for 72 hrs. CyQUANT analysis of DNA measured with a fluorescence microplate reader with filters appropriate for ~480 nm excitation and ~520 nm emission maxima. Changes in DNA fluorescence correlate with cell proliferation. C = control, CSF = cerebrospinal fluid. * P < 0.05, *** P < 0.005, **** P < 0.001, ***** P < 0.0005. b. Cells from WHO grade I (M1-M3) and WHO grade II (M4-M5) meningiomas were treated as above. * P < 0.05, *** P < 0.005, **** P < 0.001, ***** P < 0.0005.

Mentions: In the leptomeningeal cells, cerebrospinal fluid significantly stimulates DNA synthesis (p < 0.005). Co-administration of cucurbitacin I (250 nm) produces a significant blockade of this effect (p <0.0005). Cucurbitacin I alone also produced a significant reduction in basal DNA synthesis in L2 (p < 0.05) (Figure 2a). In WHO grade I meningiomas, cerebrospinal fluid significantly stimulated DNA synthesis in M1 – M3 (p < 0.001). Co-administration of cucurbitacin I (250 nm) blocked this effect (p < 0.0005). In M2, cucurbitacin I alone also significantly reduced basal synthesis (p < 0.005). In WHO grade II meningiomas, cerebrospinal fluid significantly stimulated DNA synthesis in M4 and M5 (p < 0.001). Co-administration of cucurbitacin (250 nM) significantly blocked this effect (p < 0.0005). In M5, cucurbitacin reduced basal DNA synthesis (p < 0.05) (Figure 2b).


Cucurbitacin I blocks cerebrospinal fluid and platelet derived growth factor-BB stimulation of leptomeningeal and meningioma DNA synthesis.

Johnson MD, O'Connell MJ, Walter K - BMC Complement Altern Med (2013)

Effects of cucurbitacin I on cerebrospinal fluid stimulation of leptomeningeal and meningioma cell DNA synthesis. a. Cells from 17 (L1), 20 (L2) and 22 (L3) wk fetal leptomeninges (L), were treated with CSF without or with cucurbitacin I (250 nM) for 72 hrs. CyQUANT analysis of DNA measured with a fluorescence microplate reader with filters appropriate for ~480 nm excitation and ~520 nm emission maxima. Changes in DNA fluorescence correlate with cell proliferation. C = control, CSF = cerebrospinal fluid. * P < 0.05, *** P < 0.005, **** P < 0.001, ***** P < 0.0005. b. Cells from WHO grade I (M1-M3) and WHO grade II (M4-M5) meningiomas were treated as above. * P < 0.05, *** P < 0.005, **** P < 0.001, ***** P < 0.0005.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4228312&req=5

Figure 2: Effects of cucurbitacin I on cerebrospinal fluid stimulation of leptomeningeal and meningioma cell DNA synthesis. a. Cells from 17 (L1), 20 (L2) and 22 (L3) wk fetal leptomeninges (L), were treated with CSF without or with cucurbitacin I (250 nM) for 72 hrs. CyQUANT analysis of DNA measured with a fluorescence microplate reader with filters appropriate for ~480 nm excitation and ~520 nm emission maxima. Changes in DNA fluorescence correlate with cell proliferation. C = control, CSF = cerebrospinal fluid. * P < 0.05, *** P < 0.005, **** P < 0.001, ***** P < 0.0005. b. Cells from WHO grade I (M1-M3) and WHO grade II (M4-M5) meningiomas were treated as above. * P < 0.05, *** P < 0.005, **** P < 0.001, ***** P < 0.0005.
Mentions: In the leptomeningeal cells, cerebrospinal fluid significantly stimulates DNA synthesis (p < 0.005). Co-administration of cucurbitacin I (250 nm) produces a significant blockade of this effect (p <0.0005). Cucurbitacin I alone also produced a significant reduction in basal DNA synthesis in L2 (p < 0.05) (Figure 2a). In WHO grade I meningiomas, cerebrospinal fluid significantly stimulated DNA synthesis in M1 – M3 (p < 0.001). Co-administration of cucurbitacin I (250 nm) blocked this effect (p < 0.0005). In M2, cucurbitacin I alone also significantly reduced basal synthesis (p < 0.005). In WHO grade II meningiomas, cerebrospinal fluid significantly stimulated DNA synthesis in M4 and M5 (p < 0.001). Co-administration of cucurbitacin (250 nM) significantly blocked this effect (p < 0.0005). In M5, cucurbitacin reduced basal DNA synthesis (p < 0.05) (Figure 2b).

Bottom Line: Co-administration of cucurbitacin I (250 nM) produces a significant blockade of this effect.Cucurbitacin I alone also produced a significant reduction in basal DNA synthesis.Cucurbitacin I had no effect on basal phosphorylation of STAT3 but reduced phorphorylated p44/42 MAPK in 2 grade II meningioma cells lines.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Neuropathology, Department of Pathology and Laboratory Medicine, University of Rochester School of Medicine and Dentistry, Box 626, 601 Elmwood Ave,, Rochester, NY 14642, USA. mahlon_johnson@urmc.rochester.edu.

ABSTRACT

Background: Currently, there are no consistently effective chemotherapies for recurrent and inoperable meningiomas. Recently, cucurbitacin I (JSI-124), a naturally occurring tetracyclic triterpenoid compound used as folk medicines has been found to have cytoxic and anti-proliferative properties in several malignancies thru inhibition of activator of transcription (STAT3) activation. Previously, we have found STAT3 to be activated in meningiomas, particularly higher grade tumors.

Methods: Primary leptomeningeal cultures were established from 17, 20 and 22 week human fetuses and meningioma cell cultures were established from 6 World Health Organization (WHO) grade I or II meningiomas. Cells were treated with cerebrospinal fluid from patients without neurologic disease. The effects of cucurbitacin I on cerebrospinal fluid stimulation of meningioma cell DNA synthesis phosphorylation/activation of JAK1, STAT3, pMEK1/2, p44/42MAPK, Akt, mTOR, Rb and caspase 3 activation were analyzed in human leptomeningeal and meningioma cells.

Results: Cerebrospinal fluid significantly stimulated DNA synthesis in leptomeningeal cells. Co-administration of cucurbitacin I (250 nM) produces a significant blockade of this effect. Cucurbitacin I alone also produced a significant reduction in basal DNA synthesis. In grade I and II meningiomas, cerebrospinal fluid also significantly stimulated DNA synthesis. Co-administration of cucurbitacin I (250 nM) blocked this effect.In the leptomeningeal cultures, cerebrospinal fluid stimulated STAT3 phosphorylation but not p44/42MAPK, Akt or mTOR. Cucurbitacin I had no effect on basal STAT3 phosphorylation but co-administration with cerebrospinal fluid blocked cerebrospinal fluid stimulation of STAT3 phosphorylation in each. In the grade I meningiomas, cerebrospinal fluid stimulated phosphorylation of STAT3 and decreased MEK1/2 and cucurbitacin I had no effect on basal STAT3, p44/42MAPK, Akt, JAK1, mTOR, or Rb phosphorylation. In the grade II meningiomas, cerebrospinal fluid stimulated STAT3 phosphorylation in all and reduced phosphorylation of MEK1/2 in all and p44/42MAPK in one. Cucurbitacin I had no effect on basal phosphorylation of STAT3 but reduced phorphorylated p44/42 MAPK in 2 grade II meningioma cells lines.

Conclusions: These studies raise the possibility that cucurbitacin I might have value as an adjunct chemotherapy. Additional studies are warranted to evaluate the effects of cucurbitacin I on meningiomas in vivo.

Show MeSH
Related in: MedlinePlus