ABSTRACT
Oncolytic virotherapies have emerged as new modalities for cancer treatment. We previously reported that coxsackievirus B3 (CVB3) is a novel oncolytic virus (OV) with a strong ability to lyse human non-small cell lung cancer cells; however, its non-specific toxicity against normal cells remains to be resolved. To improve its safety profile, microRNA target sequences complementary to miR-34a/c, which is expressed preferentially in normal cells, were inserted into the 5' UTR or 3' UTR of the CVB3 genome. In the presence of miR-34a/c, the gene-modified CVB3 could not replicate in normal cells. We also found that the pathogenicity of CVB3 was reduced to a greater extent by targeting miR-34a than miR-34c; in addition, it was more effective to insert the target sequences into the 3' UTR rather than the 5' UTR of the viral genome. Ultimately, we developed a double-miR-34a targeting virus (53a-CVB) by inserting miR-34a targets in both the 5' UTR and 3' UTR of the virus. 53a-CVB was minimally toxic to cells in normal tissue, but maintained nearly its full oncolytic activity in mice xenografted with human lung cancer. 53a-CVB is the first miR-34-regulated OV and represents a promising platform for the development of safe and effective anti-cancer therapies.
ABSTRACT
BACKGROUND: Shikonin is a major active chemical component extracted from Lithospermi Radix, an effective traditional herb in various types of wound healing. Shikonin can accelerate granulomatous tissue formation by the rat cotton pellet method and induce neovascularization in granulomatous tissue. The purpose of the study was to investigate its mechanism of action in human skin cells. METHODS: MTS assay was used to measure cell growth. The collagen type I (COL1 ) mRNA expression and procollagen type I C-peptide (PIP) production were detected by real-time quantitative reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. Immunofluorescence and western blot analyses were carried out to investigate nuclear factor-κB (NF-κB) signaling pathway. Cell-based proteasome activity assay was used to determine proteasome activity. RESULTS: In this study, we found that 10 µmol/L shikonin stimulated the growth of normal human keratinocytes and 1 µmol/L shikonin promoted growth of human dermal fibroblasts. However, shikonin did not directly induce COL1 mRNA expression and PIP production in dermal fibroblasts in vitro. In addition, 1 µmol/L shikonin inhibited translocation of NF-κB p65 from cytoplasm to nucleus induced by tumor necrosis factor-α stimulation in dermal fibroblasts. Furthermore, shikonin inhibited chymotrypsin-like activity of proteasome and was associated with accumulation of phosphorylated inhibitor κB-α in dermal fibroblasts. CONCLUSIONS: These results suggested that shikonin may promote wound healing via its cell growth promoting activity and suppress skin inflammation via inhibitory activity on proteasome. Thus, shikonin may be a potential therapeutic reagent both in wound healing and inflammatory skin diseases.
Subject(s)
Cell Proliferation/drug effects , NF-kappa B/metabolism , Naphthoquinones/pharmacology , Proteasome Endopeptidase Complex/drug effects , Skin/cytology , Cells, Cultured , Enzyme-Linked Immunosorbent Assay , Fibroblasts/drug effects , Humans , Keratinocytes/drug effects , Polymerase Chain ReactionABSTRACT
From the 80% acetone extract of "Cimicifugae Rhizoma" (a mixture of Cimicifuga dahurica and C. heracleifolia used medicinally), seven new fukiic acid derivatives (1-7) and a new phenylethanoid derivative (8) were isolated along with eight known compounds (9-16). Fukinolic acid (9) and cimicifugic acids A-J (10-16, 5-7) showed stronger hyaluronidase inhibitory activities than the positive control, rosmarinic acid.
Subject(s)
Caffeic Acids/isolation & purification , Caffeic Acids/pharmacology , Cimicifuga/chemistry , Hyaluronoglucosaminidase/antagonists & inhibitors , Phenylacetates/isolation & purification , Phenylacetates/pharmacology , Phenylpropionates/isolation & purification , Phenylpropionates/pharmacology , Plants, Medicinal/chemistry , Succinates/isolation & purification , Succinates/pharmacology , Caffeic Acids/chemistry , Cinnamates/chemistry , Depsides/chemistry , Japan , Molecular Structure , Nuclear Magnetic Resonance, Biomolecular , Phenylacetates/chemistry , Phenylpropionates/chemistry , Rhizome/chemistry , Succinates/chemistry , Rosmarinic AcidABSTRACT
Chemical investigation of the aerial parts of Cimicifuga simplex afforded four new fukinolic acid analogues, cimicifugic acids K-N (1-4), and 10 known compounds, and C. japonica afforded three new fukinolic acid analogues, cimicifugic acids K-M (1-3), a new phenolic glycoside, shomaside F (5), and 10 known compounds. Cimicifugic acids K-N showed more potent hyaluronidase inhibitory activities than rosmarinic acid.
Subject(s)
Caffeic Acids/isolation & purification , Cimicifuga/chemistry , Hyaluronoglucosaminidase/antagonists & inhibitors , Phenols/isolation & purification , Phenylacetates/isolation & purification , Plants, Medicinal/chemistry , Caffeic Acids/chemistry , Cinnamates/pharmacology , Depsides/pharmacology , Japan , Molecular Structure , Phenols/chemistry , Phenylacetates/chemistry , Rosmarinic AcidABSTRACT
Two polyhydroxysteroids and three steroidal saponins (pectiniosides A, B and C) were isolated as bioactive substances from Asterina pectinifera. These compounds inhibited the release of guinea-pig skin stratum corneum cells by 48-67% (control, 0%) at 2 mg/ml. It is suggested that the bioactivity of these compounds is connected with the remedial and preventive effects of an aqueous extract of A. pectinifera on rough human skin.
