Background
Methods
General details
Plant material
Extraction and isolation
Identification of 2-ethoxystypandrone (1)
Cell lines and cell culture
Reporter plasmid construction and cell transfection
STAT3-dependent luciferase reporter assay
Cell viability assay
HCC Cancer stem cells culture
Tumorsphere passage and tumorsphere formation assay
Western blot analysis
Flow cytometric analysis
Statistic analysis
Results
Isolation and structure determination
No | 13C δ (ppm) | 1H δ (ppm) | HMBC (1H → 13C) |
---|---|---|---|
1 | 178.2(C) | H-8; H-3 | |
2 | 160.4(C) | H-3; H-14 | |
3 | 109.2(CH) | 6.25 (s) | |
4 | 190.6(C) | H-3 | |
5 | 157.2(C) | ||
6 | 130.5(C) | H-12; H-13; H-8 | |
7 | 142.6(C) | H-8; H-13 | |
8 | 120.3(CH) | 7.44 (s) | H-13 |
9 | 136.0(C) | ||
10 | 112.0(C) | H-3; H-8 | |
11 | 201.6(C) | H-12 | |
12 | 30.5(CH3) | 2.55 (s) | |
13 | 18.5(CH3) | 2.35 (s) | H-8 |
14 | 65.3(CH2) | 4.22(q, J = 7.0 Hz) | H-15 |
15 | 12.9(CH3) | 1.48(t, J = 7.0 Hz) | H-14 |
Inhibitory activity of the isolated compounds against STAT3 signaling pathway
compound |
1
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2
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3
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4
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5
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6
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7
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8
| pyridone 6 |
---|---|---|---|---|---|---|---|---|---|
IC50 (μM) | 7.75 ± 0.18 | 40.20 ± 0.17 | > 100 | > 100 | > 100 | > 100 | > 100 | > 100 | 0.02 ± 0.001 |