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Volumn 20, Issue 9, 2010, Pages 2994-2997

Cucurbitacin E as a new inhibitor of cofilin phosphorylation in human leukemia U937 cells

Author keywords

Biotin linked cucurbitacin E; Cofilin; Cucurbitacin E; Cucurbitane type triterpene; Cytotoxic effect; Target protein

Indexed keywords

COFILIN; CUCURBITACIN; CUCURBITACIN E; CUCURBITACIN I; F ACTIN; G ACTIN; UNCLASSIFIED DRUG;

EID: 77950864226     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.bmcl.2010.02.062     Document Type: Article
Times cited : (62)

References (31)
  • 12
    • 77950858415 scopus 로고    scopus 로고
    • note
    • 2O (50:50, v/v)] to give 1 (31 mg, 80%).
  • 14
    • 77950859038 scopus 로고    scopus 로고
    • note
    • 15 (7 mg, 38%).
  • 16
    • 77950863188 scopus 로고    scopus 로고
    • note
    • 50 value was determined graphically.
  • 17
    • 77950864429 scopus 로고    scopus 로고
    • note
    • 50 values of cucurbitacins B and D and 23,24-dihydrocucubitacins B and E were 13 nM, 0.28 μM, 0.29 μM, and 3.1 μM in HL60 cells and 27 nM, 0.50 μM, 1.2 μM, and 9.6 μM in HT1080 cells, respectively. These results confirm the importance of a double bond at the 23,24-positions as well as an acetoxy group at the 25-position.
  • 22
    • 77950867469 scopus 로고    scopus 로고
    • note
    • 2/M phase were analyzed using cell cycle analysis software, ModFit LT ver. 3.0 (Verity Software House).
  • 23
    • 77950865242 scopus 로고    scopus 로고
    • note
    • 4 cells/mL) were incubated with the test compounds for 24 h. They were then lysed in a lysis buffer containing a phosphatase inhibitor (Roche) and a proteinase inhibitor (Thermo Scientific). Protein concentrations of cell lysate were determined using the Protein Assay Kit (Thermo Scientific). Equivalent amounts of protein (60 μg or 30 μg of protein/lane) were electrophoresed in 10% SDS-polyacrylamide gels and transferred to polyvinylidene difluoride membranes. The membranes were blocked for 30 min in Blocking One or Blocking One-P (Nacalai Tesque). The blots were probed with primary antibody at the appropriate dilution in Tris-buffered saline containing 0.1% Tween 20 (T-TBS) for 1 h. The membranes were then washed three times with T-TBS and incubated with the appropriate HRP-conjugated secondary antibody for 1 h. Immunoreactive proteins were detected using an enhanced chemiluminescence kit (ECL plus, GE Healthcare), according to the manufacturer's instructions. The following antibodies and dilutions were used for Western blotting: rabbit polyclonal antibody against anti-phospho-STAT3, anti-STAT3, anti-phospho-cofilin, anti-phospho-LIMK1/2, anti-β-actin (1:1000) (Cell Signaling Technology); anti-rabbit IgG; and HRP-linked whole antibody from donkeys (1:5000) (GE Healthcare).
  • 24
    • 77950867470 scopus 로고    scopus 로고
    • note
    • +.
  • 26
    • 77950859846 scopus 로고    scopus 로고
    • note
    • Details regarding in-gel digestion, LC-MS/MS analysis, and the Scaffold algorithm can be found in the technical information section of the website of NextGen Sciences (www.nextgensciences.com).


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.