Table 2.
(a) In Vitro Assays | ||
Source Species/Compounds | Cell Line or Purified Protein | Assays 1 and Effects Examined |
Dicathais orbita (Chloroform extracts, purified tyrindoleninone & 6 Bromoisatin) [51,68,69,104] | A range of female reproductive, colon and breast tumurs and lymphomas (Figure 3) | MTS/MTT cell viability; Crystal violet; Caspase 3/7 activity for apoptosis; Lactate dehydrogenase for necrosis; Tunnel staining for apoptosis; Flow cytometry for apoptosis, necrosis and cell cycle analysis |
Dicathais orbita (extracts and compounds) [69,106] | JAr and human granulosa cells | Radioimmunoassays (RIA); Steroidogenesis assays: estradiol (E2) and progesterone (P4) synthesis |
Hexaplex trunculus (purified bromoindirubins) [107,108] | Recombinant or naturally purified protein kinases | CDK1/Cyclin B, CDK5/p25, GSK-3 and other protein receptor kinase assays |
Rapana venosa (Ethanol extracts) [109] | Human leukemia HL-60 and human lung cancer A-549 | MTT cell viability and liquid-scintillation radioassay for cell proliferation (3H-TdR) |
Thais clavigera (Ethanol extracts) [109] | Human leukemia HL-60 and human lung cancer A-549 | MTT cell viability and liquid-scintillation radioassay for cell proliferation (3H-TdR) |
Rapana thomasiana (Purified haemocyanin) [110] | SiHa-cervical squamous cell carcinoma, CaOV-ovarian adenocarcinoma, MIA PaCa-pancreatic carcinoma, RD 64-rhabdomyosarcoma, EJ-urinary bladder carcinoma and Lep-nontumor human lung cell line. | Cell proliferation assay and apoptosis indicated by DNA degradation and caspase-3 activation |
Rapana venosa (Haemocyanins) [111] | 647-V, T-24 and CAL_29 bladder cancer cells | MTT AND WST-1 cell viability assays, apoptosis with acridine orange/propidium iodine staining and gene expression profiles for 168 inflammatory cytokines and signal transduction pathways. |
Synthetic isatin derivatives [105,112] | The human leukemic (U937, monocyte and Jurkat, T cell), breast (MDA-MB-231 and MCF-7), prostate (PC-3), and colorectal (HCT-116) | MTS cell viability, caspase 3/7 for apoptosis, CDK2 inhibition |
Synthetic indirubin derivatives [107,108,113,114,115,116] | Recombinant or naturally purified protein kinases | CDK1/Cyclin B, CDK5/p25, GSK-3 and other protein receptor kinase assays; affinity chromatography; crystallography and in silico modelling; rt PCR on Xenopus embryos |
Synthetic indirubin derivatives [117,118] | Human neuroblastoma and breast cancer cell lines | Apoptosis induction pathways |
Synthetic indirubin derivatives [119,120] | Human melanoma andmyeloid leukemia cell lines | Jak/Stat 3 phosphorylation, FLT3 inhibition |
Synthetic isatin and indirubin derivatives [69,106,121] | JAr and human granulosa cells for female hormones and H294 adrenal cells for male | RAI; ELISA for E2 and P4; E-screen (xeno-oestrogenic potential) for E2 receptor binding; H294 adrenal cells for cortisol, testosterone, androgen, and didehydroepiandrosterone |
Synthetic indirubin derivatives [122] | JAr and human granulosa cells | RAI |
(b) In Vivo Models | ||
Source Species/Compounds | Cancer Type | Animal Model |
Dicathais orbita (Chloroform extracts) [123], (purified tyrindoleninone, 6 bromoisatin) [124] | Colon cancer prevention | Apoptotic response to genotoxic damage by azoxymethane (AOM) in mice. Compounds delivered by oral gavage two weeks prior to AOM |
Concholepas concholepas (haemocyanin subunits CCHA & CCHB) [92] | Bladder carcinoma treatment | MBT-2 heterotopic murine bladder carcinoma model |
Synthetic 6-bromoistain [112] | Colon cancer prevention | Apoptotic response to genotoxic damage by AOM in mice. Compound delivered by oral gavage two weeks prior to AOM |
Synthetic 6-bromoindirubin derivatives [119] | Human melanoma treatment | Xenograph model in BALC/c mice, 14 day treatment |
Synthetic indirubin derivatives [125] | Renal, prostate, lung and colon cancer treatment | Xenograph model in BALC/c mice |
1 MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt), MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and WST-1 (4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate) are tetrazolium reduction assays for cell viability.