Table 1.
Studies on the inhibition of EMT and stemness by PTs.
| PT | Experimental Model | Conditions | Biological Effects *** | Effects on Cell Signalings **** | Refs. | ||
|---|---|---|---|---|---|---|---|
| Type | Name | Conc. * | Time ** | ||||
| Ursane | Asiatic acid (AA) | A549 treated with TGF-β1 (10 ng/mL) | 10–40 | 24 h | ↓ morphological changes (20 μM AA), ↓ migration DD, ↓ invasion DD | ↓ β-catenin DD, ↓ p-GSK-3β DD, ↑ E-cadherin DD, ↓ N-cadherin DD, ↓ vimentin DD, ↓ Snail DD | [164] |
| Pulmonary fibrosis in C57BL/6 mice was induced by single intratracheal administration of bleomycin (3 mg/kg) | 5–20 mg/kg/day (intragastrically) | 21 d | ↓ collagen deposition DD, ↓ histopathological changes in the lungs DD, ↑ pulmonary function DD, ↓ macrophage, neutrophil and lymphocyte infiltration in BALF DD | ↓ collagen I DD, ↓ collagen III DD, ↓ α-SMA DD, ↓ TIMP-1 DD, ↓ vimentin DD, ↑ E-cadherin DD, ↓ TGF-β1 DD, ↓ p-Smad2/3 DD, ↓ p-ERK1/2 DD, ↓ IL-1β DD, ↓ IL-18 DD, ↓ IL-6 DD, ↓ TNF-α DD, ↓ NLRP3 DD, ↓ ASC DD, ↓ pro-Caspase-1 DD, ↓ active Caspase-1 DD | [165] | ||
| Ursane | Ursolic acid (UA) | A549, H1975 | 5–30 | 24 h | ↓ adhesion to Matrigel DD (A549), ↓ migration DD (A549, H1975), ↓ invasion DD (A549, H1975) | ↑ E-cadherin DD (A549), ↓ N-cadherin DD (A549), ↓ vimentin (A549, 20 μM UA); 84 genes regulated by UA (A549, 30 μM UA) were associated with the signaling pathways of TGF-β, ECM-receptors, adherens junctions, Wnt, VEGF, tight junctions, cell adhesion molecules; ↓ AEG-1 DD,K (A549) | [166] |
| A549 treated with TNF-α (5 ng/mL) | 5–20 | 12/24 h with UA, then 12/24 h with TNF-α | ↓ NF-κB p65 subunitK (20 μM UA, 12 h), ↓ AEG-1 DD (24 h) | ||||
| H1975 treated with TGF-β1 (5 ng/mL) | 0.02 | 24 h | ↓ morphological changes, ↓ migration, ↓ invasion | ↑ E-cadherin, ↓ N-cadherin, ↓ MMP-2 catalytic activity, ↓ MMP-9 catalytic activity, ↓ MMP-2, ↓ MMP-9, ↓ integrin αVβ5K | [167] | ||
| HBE treated with 1% cigarette smoke extract (CSE) | 10 | 2 h prior to CSE | ↓ TGF-β1, ↓ p-Smad2/3, ↓ S100A4, ↑ IGF-1 | [168] | |||
| Emphysema in Wistar rats was induced by exposure to cigarette smoke for 30 min, two times a day, 6 days a week, for 3 months | 10–40 mg/kg/day (intragastrically) | 3 mos | ↓ airway-vessel remodeling, ↓ collagen deposition, ↓ mucus secretion | ↓ α-SMA DD, ↓ S100A4, ↓ TGF-β1, ↓ p-Smad2/3, ↑ IGF-1 DD | |||
| Emphysema in SD rats induced by exposure to cigarette smoke for 30 min, two times a day, 6 days a week, for 3 months | 10–40 mg/kg/day (intragastrically) | 3 mos | ↓ p-IRE1, ↓ XBP1 | [169] | |||
| Emphysema in SD rats induced by intraperitoneal injection of CSE on days 1, 8, 15, 21 | 20 mg/kg/day (intragastrically) | 2–4 wk | ↓ airway remodeling | ↓ p-Smad2/3, ↓ p-PERK, ↓ PERK, ↓ p-eIF-2α, ↓ e-IF-2α, ↓ ATF4, ↓ CHOP, ↓ p-IRE1, ↓ ATF6, ↓ active Caspase 12 | |||
| A549, H460 | 10, 20 | 1–14 d | ↓ migration DD, ↓ invasion DD, ↓ tumorsphere formation (20 μM UA; 7 d, 14 d) | ↓ VEGF DD (24 h), ↓ NANOG (tumorpheres; 20 μM UA; 24 h), ↓ OCT4 (tumorpheres; 20 μM UA; 24 h), ↓ SOX2 (tumorpheres; 20 μM UA; 24 h), ↓ pEGFR (24 h), ↓ pJAK2 DD (24 h), ↓ pSTAT3 DD (24 h), ↓ PD-L1 DD (24 h), ↓ MMP2 DD (24 h), ↓ MMP3 DD (24 h), ↓ MMP9 DD (24 h), ↓ VEGF DD (24 h), ↓ the binding of STAT3 to MMP2 and PD-L1 promoters (20 μM UA, 24 h) | [170] | ||
| A549 and H460 treated with EGF (25 ng/mL) | 20 | 1 h with EGF, then 24 h with UA | ↓ pEGFR | ||||
| H1975 harbouring L858R/T790M mutation | 25 | 12–72 h | ↓ migration | ↓ CT45A2K (12 h), ↓ the binding of TCF4 to CT45A2 promotorK (12 h), ↓ TCF4 (48 h), ↓ p-β-catenin (48 h), ↑ p-GSK-3β (48 h), ↓ nuclear translocation of β-catenin | [171] | ||
| subcutaneous injection of H1975 in athymic nude mice | 25 mg/kg/day | 18 d | ↓ CT45A2, ↓ p-β-catenin, ↑ p-GSK-3β, ↓ TCF4 | ||||
| Ursane | Ursonic acid | A549, H1299 | 2.5, 5 | 24 h, 48 h | ↓ invasion (24 h) | ↓ MMP-2 catalytic activity DD (A549, H1299; 48 h), ↓ MMP-9 catalytic activity (H1299, 48 h), ↓ MMP-2 DD (A549, H1299; 48 h), ↓ MMP-9 DD (H1299, 48 h), ↓ RECK DD (A549, 48 h), ↑ RECK (H1299, 48 h), ↓ p-ERK DD (A549, H1299; 24 h), ↓ p-CREB DD (A549, H1299; 24 h) | [172] |
| Oleanane | C DDO-Me | Radiation-induced lung inflammation in C57BL/6 mice was induced by thoracic irradiation with a single dose of 12.5 Gy | 600 ng intragastrically on days -1, 1, 3 and 5 | 3 wk | ↓ inflammatory cells infiltration in BALF, ↓ total protein in BALF, ↓ histopathological changes in the lungs | ↓ IL-6, ↓ TGF-β, ↑ IL-10, ↓ fibronectin, ↓ α-SMA, ↓ collagen I | [173] |
| Radiation-induced pulmonary fibrosis in C57BL/6 mice was induced by thoracic irradiation with a single dose of 22.5 Gy | 600 ng intragastrically on days -1, 1, 3, 5, 7 and 9 | 12 wk | ↓ collagen deposition | ↓ fibronectin, ↓ α-SMA, ↓ collagen I | |||
| Friedelane | Celastrol | A549 treated with TGF-β1 (5 ng/mL) | 1 | 30 min with celastrol, then 24 h–72 h with TGF-β1 | ↓ morphological changes (72 h), ↓ invasion (36 h) | ↑ E-cadherin (72 h), ↓ Snail (24 h) | [152] |
| A549 treated with TGF-β1 (5 ng/mL) | 5 | 24 h | ↑ E-cadherin, ↑ ZO-1, ↓ N-cadherin, ↓ vimentin, ↓ Snail, ↓ Slug | [174] | |||
| Pulmonary fibrosis in Wistar albino rats was induced by single intratracheal administration of bleomycin (3 U/kg) | 5 mg/kg, twice a week | 28 d | ↓ TGF-β1, ↓ p-Smad2/3, ↑ E-cadherin, ↑ claudin, ↓ N-cadherin, ↓ Snail, ↓ Slug, ↓ β-catenin, ↓ Hsp90 | ||||
| Oleanane | Evoditrilone A | A549 | 1–2 | 48 h | ↓ colony formation ability DD, ↓ migration DD | ↑ E-cadherin DD, ↓ MMP-2 DD, ↓ N-cadherin DD | [148] |
| Oleanane | Glycyrrhizin | A549 and BEAS-2B treated with TGF-β1 (5 ng/mL) | 50–200 (A549); 25–100 (BEAS-2B) |
2 h with glycyrrhizin, then 24 h with TGF-β1 | ↓ migration DD | ↓ HMGB1 secretion DD, ↓ HMGB1 DD, ↓ p-Smad2/3 DD, ↑ E-cadherin DD, ↓ vimentin DD | [149] |
| A549 and BEAS-2B with lentivirus-mediated HMGB1 overexpression | 100 (A549); 50 (BEAS-2B) | 24 h | ↓ HMGB1 secretion, ↓ HMGB1, ↓ TGF-β1, ↓ p-Smad2/3, ↑ E-cadherin, ↓ vimentin | ||||
| Oleanane | Oleanolic acid (OA), OA-loaded P105/TPGS mixed micelles | A549, PC-9 | 15, 30 | 24 h | ↓ migration (OA-micelles > free OA), ↓ invasion (OA-micelles > free OA) | ↑ E-cadherin (OA-micelles > free OA), ↓ N-cadherin (OA-micelles > free OA), ↓ p-ERK (OA-micelles > free OA) | [175] |
| Friedelane | Pristimerin (Pr) | H1299 | 0.9–3.6 | 24–72 h | ↓ colony formation ability TD (1.8 μM Pr), ↓ migration TD,DD, ↓ invasion DD (48 h) | ↓ vimentin (3.6 μM Pr, 48 h), ↓ F-actin (0.9–3.6 μM Pr, 48 h), ↓ integrin β1 (3.6 μM Pr, 48 h), ↓ MMP-2 (0.9–3.6 μM Pr, 48 h), ↓ Snail (0.9–3.6 μM Pr, 48 h) | [151] |
| Oleanane | Soloxolone methyl | A549 treated with TGF-β1 (50 ng/mL) | 0.5 | 24, 48 h | ↓ morphological changes, ↓ migration (24 h, 48 h), ↓ invasion (48 h) | ↑ E-cadherin (48 h), ↑ ZO-1 (48 h), ↓ vimentin (48 h), ↓ fibronectin (48 h) | [176] |
| Oleanane | β-escin (β-Es) | H460 | 5–40 | 24 h | ↓ ALDH+ cell population (5–40 μM β-Es), ↑ p21 (20 μM β-Es; both in ALDH+ and ALDH- cells) | [177] | |
| lung tumors in A/J mice were induced by single intraperitoneal injection of tobacco carcinogen NNK (10 μmol/mouse) | 3 weeks after NNK treatment mice were fed with the diet containing 500 ppm β-Es | 17, 34 wk | ↓ lung tumor formation, ↓ progression of adenomas to adenocarcinomas | ↑ p21 (34 wk), ↓ ALDH1A1 (34 wk), ↓ p-Akt (34 wk) | |||
| Oleanane | β-peltoboykinolic acid (β-P) | A549 treated with TGF-β1 (2 ng/mL) | 1–10 μg/mL | 24–48 h | ↓ morphological changes (5 μg/mL β-P, 10 μg/mL β-P; 48 h), ↓ migration DD (24 h, 36 h) | ↑ E-cadherin DD (48 h), ↓ N-cadherin (10 μg/mL β-P, 48 h), ↓ vimentin DD (48 h), ↓ fibronectin DD (48 h), ↓ collagen I DD (48 h), ↓ p-Smad2 (10 μg/mL β-P, 48 h), ↓ Snail DD (48 h) | [178] |
| Lupane | Betulinic acid | 293T, A549, H1299 | 10–30 | 4 h–7 d | ↓ migration (A549, H1299; 20 μM BA > 10 μM BA; 24 h), ↓ invasion (A549, H1299; 20 μM BA > 10 μM BA; 24 h), ↓ the sphere-forming ability (A549, H1299; 20 μM BA > 10 μM BA; 7 d) | the direct binding to Skp2 by forming H-bonds with Lys145, ↓ Skp2-Skp1 interactions (exogenous Flag-Skp1 was transfected in 293T, endogenous Skp2-Skp1 interactions in H1299; 20, 30 μM BA; 4 h), ↓ Skp2-mediated ubiquitination of p27 (exogenous p27 in 293T, endogenous p27 in A549; 10, 20 μM BA; 24 h), ↓ Skp2-mediated ubiquitination of E-cadherin (exogenous E-cadherin in 293T, endogenous E-cadherin in A549; 10, 20 μM BA; 24 h), ↓ Skp2 DD,TD (A549, H1299), ↑ p27 DD,TD (A549, H1299), ↑ E-cadherin DD,TD (A549, H1299) | [179] |
| intravenous injection of A549 into BALB/C nude mice (metastasis model) | 50 or 75 mg/kg BA was administered on the day 7 after LLC injection | 2 mos | ↓ metastasis DD | ||||
| A549 and H1299 treated with TGF-β1 (10 ng/mL) | 10–20 | 24 h | ↑ E-cadherin DD, ↓ vimentin (A549: 20 μM; H1299 DD), ↓ N-cadherin DD, ↓ Skp2 DD | ||||
| LLC | 10–20 | 24 h | ↓ migration DD, ↓ invasion DD | ↓ Skp2 DD, ↑ E-cadherin DD | |||
| subcutaneous injection of LLC into C57BL/6 mice (spontaneous metastasis model) | 50 or 75 mg/kg each day after LLC injection | 21 d | ↓ primary tumor growth DD, ↓ lung metastasis DD | ↓ Skp2 DD, ↑ E-cadherin DD, ↑ p27 DD | |||
| intravenous injection of LLC into C57BL/6 mice (metastasis model) | 50 or 75 mg/kg BA was administered on the day 7 after LLC injection | 60 d | ↓ lung metastasis DD | ↓ Skp2, ↑ E-cadherin | |||
| Lupane | Betulinic acid, SYK023 | H1299 | 0.1–30 | 36 h | ↓ migration (BA: 10 μM > 5 μM; SYK023 DD), ↓ invasion (BA: 10 μM > 5 μM; SYK023 DD) | ↓ F-actin polymerization (BA: 10 μM > 5 μM; SYK023 DD), ↓ p-FAK (SYK023: 1 μM, 5 μM), ↓ p-Src (BA: 5 μM > 1 μM; SYK023: 0.5–5 μM), ↓ p-Akt (BA: 5 μM; SYK023: 0.5–5 μM), ↓ p-mTOR (SYK023: 1 μM, 5 μM), ↓ N-cadherin (BA: 30 μM; SYK023 DD), ↓ β-catenin (BA: 30 μM; SYK023 DD), ↓ vimentin (SYK023: 20 μM, 30 μM), ↓ c-Myc (BA: 20 μM, 30 μM; SYK023: 20 μM, 30 μM)), ↓ SYPD K (BA, SYK023: 20 μM) | [180] |
* Concentrations are shown in μM unless otherwise stated. ** Time is given in hours (h), days (d), weeks (wk) and months (mos). *** Downward (↓) and upward (↑) arrows indicate suppression and activation of biological process, respectively. **** Downward (↓) and upward (↑) arrows indicate downregulation and upregulation of expression, respectively. DD Dose-dependent increase in effect. TD Time-dependent increase in effect. K Demonstrated as a key mechanism using pharmacological inhibition and/or genetic engineering methods.