Table 2.
In vivo evaluation of Bufalin in mouse tumor models
| Cancer type | Drugs | Animal | Tumor models | Transplantation | Treatment | Results | Refs. | Year |
|---|---|---|---|---|---|---|---|---|
| Bone | Bufalin | Athymic nude mice; male; 7–8 weeks old | Rat breast sarcocarcinoma Walker 256 cell | Inoculate 2 × 105 cell into the intramedullary space of the mouse femur | 1 day after tumor inoculation, given by i.p. 0.5, 1, 1.5, or 2 mg/kg/day for 21 days | Cancer-induced pain relief; 50% reduction in bone tissue injury | Ji et al. [83] | 2017 |
| Cervical | Bufalin | BALB/c nude mice; female; 4–5 weeks old | Human cervical squamous cell carcinoma Siha cell | Subcutaneously inoculate 3 × 106 cell | After reaching an average tumor volume of 100 mm3, given by i.p. Bufalin 10 mg/kg ± paclitaxel (10 mg/kg per 4 days) for 32 days | Bufalin synergizes with paclitaxel to inhibit tumor growth without apparent loss of body weight | Liu et al. [80] | 2016 |
| Breast | Bufalin-NP | SCID mice; female; 6–7 weeks old | Human breast MDA-MB-231-LM3.3 cell | Inject 0.75 × 106 cell into one of the second mammary fat pads | 6 days after tumor injections, given by i.v. Bufalin-NP (1.5 mg/kg) three times per week for 20 days | Bufalin sensitizes cancer cells to MK-2206 and blocks tumor growth | Wang et al. [56] | 2014 |
| Bufalin-BCS-NP | Balb/c nude mice; female; 4 weeks old | Human breast MCF-7 cell | Subcutaneously implant 1 × 107 cell into the right axilla skin | 7–10 days after tumor inoculation, (1) given by i.p. BF-BCS-NPs (1 mg/kg BF equivalent in PBS) or free BF (1 mg/kg in ethanol) every 2 days for 20 days; (2) give by i.v. BF-BCS-NPs (5 mg/kg BF equivalent in PBS) at D1, D3 | Free BF suppresses tumor growth accompanying with a significantly decreased body weight. BF-BCS-NPs suppresses tumor growth without apparent loss of body weight | Tian et al. [104] | 2014 | |
| 3-Phospho-bufalin | Nude mice; female; 4–6 weeks old | Human breast MDA-MB-231-LM3-3 cell | Inject 0.75 × 106 cell into one of the second mammary fat pads | 14 days after injection, given by s.c. phospho-BF (0.75 mg/kg per dose, 3 doses per week) for 3 weeks | 2.4 times reduction in tumor weight; no cardiotoxicity observed | Song et al. [55] | 2015 | |
| Bufalin | Athymic nude mice; female; 5 weeks old | Human breast MDA-MB-231 cell | Subcutaneously inject 5 × 106 into both dorsal flank regions | 4 weeks after injection, (1) given by intra-tumoral injection BF 10 μL (1 mM, in 0.9% normal saline) to the left flank for 4 weeks; (2) 10 μL volume of normal saline to the right flank for 4 weeks | Significantly enhances breast cancer xenograft growth; promote the inflammatory response | Chen et al. [38] | 2017 | |
| Colorectal | Bufalin-loaded mPEG-PLGA-PLL-cRGD nanoparticles (BNPs) | Athymic nude mice; female; 4–6 weeks old | Human colon SW620 cell | Inject 5 × 106 cell into the dorsal subcutaneous space | Bufalin-loaded mPEG-PLGA-PLL-cRGD nanoparticles (BNPs) containing 1 mg/kg bufalin, given by injection through the vena caudalis once a day for 14 days | Suppresses tumor growth; bufalin-loaded NPs significantly enhanced treatment efficacy compared to that of a bufalin water solution | Yin et al. [105] | 2012 |
| Bufalin | Male athymic nude mice | HCT116-luc-vector and HCT116-luc-miR-497 | Inject 1 × 106 cells intravenously via the tail vein | 1 week after injection, given by Bufalin for 1 mg/kg via the tail vein (three times a week) for 5 weeks | Inhibited colorectal cancer metastasis; improved life of survival. Improved physiological characteristics in terms of body weight, skin roughness, mental status, and survival rate | Qiu et al. [130] | 2014 | |
| Bufalin-loaded pluronic polyetherimide nanoparticles | Male athymic nude mice | HCT116 | Injecte 1 × 106 cells intravenously via the tail vein. | 2 weeks after injection, (1) given by of Bufalin 1 mg/kg; (2) given by 20 mg/kg of Bufalin-loaded pluronic PEI nanoparticles via the tail vein (0.2 mL per mouse, three times per week) for 3 weeks | Inhibited colorectal cancer metastasis. Improved quality of life and physiological characteristics in terms of body weight, skin roughness, and mental status | Hu et al. [93] | 2014 | |
| Bufalin | BALB/c mice; male; 5–6 weeks old | HCT116 | Inject 2 × 1010 cells s.c. into the right axillary region. Tumor mode of the second generation: 2 weeks after injection, harvest the s.c. xenogra tumors, cut into pieces (1.5 mm in diameter), implant into the axillary region s.c. Orthotopic xenogra model: harvest third generation s.c. tumors and cut into pieces (1.5 mm in diameter) | NS group (treated with 0.2 mL normal saline); 5-Fu group (treated with 5-FU, 25 mg/kg); low Bufalin group (0.5 mg/kg); medium Bufalin group (1.0 mg/kg); high Bufalin group (1.5 mg/kg). NS, 5-FU, and Bufalin were administrated by intraperitoneal injection, once per day from day 15 to day 21 (12 mice in each group) | Inhibit cell growth. Lower tumor volume. Prolong survival time | Wang et al. [131] | 2015 | |
| Bufalin-DOX | Athymic nude mice (BALB/c-nu/nu) of 6–8 weeks | HCT8/ADR | Inject 1 × 107 cells s.c. under the shoulder in the nude mice | Mice were randomized into six groups (6 in each group) when the tumor volumes reached 150–200 mm3: control; BF (0.1 mg/kg, i.p., q3d × 5); DOX (0.1 mg/kg, i.p., q3d × 5); DOX (0.5 mg/kg, i.p., q3d × 5); DOX (1.0 mg/kg, i.p., q3d × 5); DOX (0.1 mg/kg, i.p., q3d × 5) plus BF (0.1 mg/kg, i.p., q3d × 5, given 1 h before DOX administration) | BF remarkable increased the effect of DOX against the ABCB1 resistant HCT8/ADR colorectal cell xenografts in nude mice | Yuan et al. [145] | 2015 | |
| Bufalin | Male nude mice (BALB/c nu/nu, 5-week-old) | HCT116 | Inject 2 × 106 cells into the subcutaneous tissues. | 2 weeks after injection, (1) given by cisplatin (10 mg/kg body weight) i.p. every 3 days for 4 weeks.; (2) given by bufalin (1 mg/kg body weight) i.p. every 3 days for 4 weeks | Inhibition of tumor growth and tumor tissue weights are greater with the combination of cisplatin and bufalin than with cisplatin alone. Tumors treated with the combination of cisplatin and bufalin showed more cell vacuolization and nuclear shrinkage than with cisplatin alone | Sun et al. [146] | 2017 | |
| Gallbladder | Bufalin | Male athymic nude mice (5 week-old) | GBC-SD | Xenograft model: Inoculate 1 × 106 GBC-SD cells into the left axillary region | 24 h postinoculation, given by PBS i.p. and bufalin with (0.1, 0.2, and 0.4 mg/kg) i.p. every 2 days for up to 20 days | Suppression of tumor growth | Jiang et al. [45] | 2014 |
| Liver | Bufalin | BALB/c nu/nu mice (18–20 g, 5 week-old) | HCCLM3 | Inject 5 × 106 cells s.c. into the upper left flank region of nude mice. Revmove the tumors when reached approximately 1 cm in length (approximately 4 weeks after injection) and mince into small pieces of equal volume (1.5–2 × 2 × 2 mm3), then transplant into the livers of 24 nude mice | From day 8 to 38, given by1 mg/kg Bufalin; 1.5 mg/kg Bufalin; 100 mg/kg LY294002 and saline i.p. thrice weekly, respectively | 1.5 mg/kg Bufalin decreased the sizes and qualities of orthotopic transplanted tumors as well as pulmonary metastasis. Orthotopic transplanted tumor tissues were necrotic and the apoptotic cell number was markedly higher in 1.5 mg/kg Bufalin group. Inhibition of AKT/GSK3β/β-catenin/E-cadherin signaling pathways | Zhang et al. [72] | 2014 |
| Bufalin–sorafenib | BALBc nu/nu mice (6 week-old) | Human HCC cell lines SMMC7721 | Inoculate 5 × 106 cells s.c. into the abdominal intraderma | Control group: inject with the vehicle i.p. (5 days/week, 2 weeks). Experimental group: (1) injection of 1 mg/kg bufalin i.p. (5 days/week, 2 weeks); (2) oral uptake of 30 mg/kg/day sorafenib (5 days/week, 2 weeks); (3) the combination of both injections of bufalin i.p. and oral uptake of sorafenib (5 days/week, 2 weeks) | Inhibit blood vessel formation in the intradermal tumors, manifested by the vessel numbers and branches and attenuate tumor weight in nude mice with the combination treatment | Wang et al. [64] | 2016 | |
| Bufalin | BALBc nu/nu mice (6 week-old) | Human HCC cell lines SMMC7721 | Inoculate 5 × 106 cells s.c. into the right flank | Tumor size was measured every 4 days after the treatment. Tumor-bearing mice were sacrificed after 16 days of treatment, and the tumor weight was evaluated | Combination treatment inhibits tumor growth and tumor angiogenesis in vivo. The combination treatment group showed more reduced microvessel density than any other group |
Wang et al. [64] | 2016 | |
| Bufalin | BALBc nu/nu mice (6 week-old) | SMMC7721-GFP | Inoculate 5 × 106 cells s.c. into the right flanks. 4 weeks later, cut the non-necrotic tumor tissue into 1 mm3 pieces and orthotopically implanted into the liver. In addition, inject 2 × 106 via mouse tail veins | (1) Inject 1 mg/kg bufalin i.p. (5 days/week for six weeks); (2) Inject PBS i.p. (5 days/week for 6 weeks) | Bufalin reduced liver/lung metastases. Bufalin inhibited invasion through EMT | Wang et al. [76] | 2016 | |
| Myeloma | Bufalin-MK2206 | BALB-c nu/nu female mice (4–6 week-old); NOD-SCID female mice (4–6 week-old) | MOPC315; H929 | Inject 2 × 107 MOPC315 cells s.c. in the right flanks of the BALB-c nu/nu mice. Inject 1 × 107 cells H929 cells s.c. in the right hind leg of NOD/SCID mice | Mice bearing MOPC315 MM tumors were treated with bufalin (1 mg/kg; intraperitoneally) daily in the presence and/or absence of MK2206 (120 mg/kg orally) for 10 days. Mice injected with H929 MM cells were treated with 1 mg/kg bufalin daily with or without 120 mg/kg MK2206 for 12 days | Bufalin combined with MK2206 blocked MM tumor growth, decreased tumor cell proliferation and increased the percentage of apoptotic cells | Xiang et al. [147] | 2017 |
| Osteosarcoma | Bufalin | BALB/c nude mice; female; 6 week-old | U2OS/MTX300 | Inject 5.6 × 106 cells s.c. into the axilla of the mice | 10 days after injection, (1) control group: 100 mL of vehicle i.p.; MTX group: MTX (250 mg/kg) with calcium leucovorin rescue (24 mg/kg at 16, 20, or 24 h after MTX) i.p. per week; (3) low Bufalin group: 0.75 mg/kg i.p.; (4) High Bufalin group:1.5 mg/kg i.p. | Bufalin inhibited tumor growth | Xie et al. [86] | 2012 |
| Pancreas | Bufalin | BALB/c nu/nu mice; male; 4-week-old | Mia PaCa-2 | Inject 6 × 106 cells s.c. into the back of mice | When tumors reached the size of 100 mm3: (1) vehicle alone (control); (2) Bufalin (0.1 mg/kg, for 10 days); (3) Gemcitabine (125 mg/kg, 3 times/week for 2 weeks); (4) Bufalin and Gemcitabine in combination | Bufalin potentiates the anti-tumor effect of gemcitabine in vivo. Combination treatment with gemcitabine signicantly reduced the tumor volume and cell proliferation activity | Chen et al. [148] | 2012 |
| Bufalin | BALBc nu/nu mice; female; 6-week-old | MiaPaCa2/GEM | Inoculated 2 × 105 cells into the right flanks of mice | Given by (1) injections of 1.5 mg/kg bufalin (5 days/week) for 4 weeks; (2) injections with vehicle (20 μL saline) for 4 weeks. In addition, MiaPaCa2/GEM cells (2 × 106) was given to one of another two groups pre-treated with bufalin via the tail veins for 6 weeks | Inhibit pancreatic tumor growth | Wang et al. [82] | 2016 | |
| Bufalin | BALBc nude mice; male; 5-week-old) | BxPC3-luc2 | Injected 1 × 107 cells s.c. into the left buttock of mice | 7 days after inoculation, control group: inject saline i.p.; Bufalin groups: inject 1 mg/kg and 2 mg/kg for 14 days; Positive control: DDP 2 mg/kg every other day i.p. for 14 days | Bufalin treatment inhibits tumor growth | Liu et al. [149] | 2016 | |
| Lung | Bufalin | BALB/c nu/nu mice; male; 6–8 week-old | NCI-H460 | Inject 1.3 × 107 cells s.c. into flank of each mouse | When tumor volume exceeded 100 mm3, given by (1) vehicle (0.1% DMSO); (2) Bufalin: 0.1, 0.2, or 0.4 mg/kg for 14 days | Bufalin suppresses tumor growth | Wu et al. [150] | 2017 |
NP nanoparticles, BSC biotin modified chitosan, s.c. subcutaneously, i.p. intraperitoneally