Homeopathic mother tincture of Phytolacca decandra induces apoptosis in skin melanoma cells by activating caspase-mediated signaling via reactive oxygen species elevation

OBJECTIVE： Preventive measures against skin melanoma like chemotherapy are useful but suffer from chronic side effects and drug resistance. Ethanolic extract of Phytolacca decandra （PD）, used in homeopathy for the treatment of various ailments like chronic rheumatism, regular conjunctivitis, psoriasis, and in some skin diseases was tested for its possible anticancer potential. METHODS： Cytotoxicity of the drug was tested by conducting 3-（4,5-dimethylthiazol-2-yl）-2,5- diphenyltetrazolium bromide assay on both normal （peripheral blood mononuclear cells） and A375 cells. Fluorescence microscopic study of 4＇,6-diamidino-2-phenylindole dihydrochloride-stained cells was conducted for DNA fragmentation assay, and changes in cellular morphology, if any, were also recorded. Lactate dehydrogenase activity assay was done to evaluate the percentages of apoptosis and necrosis. Reactive oxygen species （ROS） accumulation, if any, and expression study of apoptotic genes also were evaluated to pin-point the actual events of apoptosis. RESULTS： Results showed that PD administration caused a remarkable reduction in proliferation of A375 cells, without showing much cytotoxicity on peripheral blood mononuclear cells. Generation of ROS and DNA damage, which made the cancer cells prone to apoptosis, were found to be enhanced in PD-treated cells. These results were duly supported by the analytical data on expression of different cellular and nuclear proteins, as for example, by down- regulation of Akt and Bcl-2, up-regulation of p53, Bax and caspase 3, and an increase in number of cell deaths by apoptosis in A375 cells. CONCLUSION： Overall results demonstrate anticancer potentials of PD on A375 cells through activation of caspase-mediated signaling and ROS generation.

Effects of Nicotinamide on Mouse Skin Tumor Development and Its Mode of Action

Nicotinamide (NA), a naturally occuring vitamin and a protease inhibitor, has been shown to be effective in treating some skin ailments. It inhibits cell proiferaion and induces cell differentiation. This report shows the effects of NA on mouse skin tumor development and on the critical events involved in this process. NA reduced tumor growth, inhibited the 12-O-tetradecanoylphorbol-13-acetate (TPA) induced ornithine decarboxylase activity, but induced the transglutaminare activity which was inhibited by TPA under different experimental conditions.The effects of NA on ornithine decarboxylare (ODC) and transglutaminase (TG) indicated that nicotinamide (NA) probably programmmed the cells for their death in the natural course of time, i.e. programed cell death. This observation indicates that NA might be a better agent for the detailed study and for the better use in prevention of cancer alone or in combination with other drugs.

外鼻皮肤癌的冷冻治疗临床观察

外鼻皮肤癌好发于老年人,多为基底细胞癌和鳞状细胞癌,其特点为原发性、恶性程度低、发展较慢且易于发现,目前的治疗多以外科治疗为主,但修复根治性切除后所造成的鼻部缺损,常需要皮瓣的修复重建鼻部外形,损伤较大,老年人不易耐受,我科自2005年应用自制冷冻器实施液氮冷冻治疗此病,不需要皮瓣修复,减少了患者的创伤,取得了良好的治疗效果。

Evaluation of chemopreventive potentials of ethanolic extract of Ruta graveolens against A375 skin melanoma cells in vitro and induced skin cancer in mice in vivo

OBJECTIVE： Chemopreventive approach with natural products, particularly plants and plant-derived ones, is receiving increasing attention for their effective role against cancer without any palpable side effects. In this study, efficacy of ethanolic extract of Ruta graveolens （RG） on skin melanoma cells （A375） in vitro and on 7,12-dimethylbenz（a）anthracene （DMBA）-induced skin cancer in vivo has been tested in Swiss albino mice. METHODS： Studies on cell viability, apoptosis and autophagy induction were conducted in vitro. To check apoptosis, assays like alteration in mitochondrial membrane potential, annexin V-fluorescein isothiocyanate/propidium iodide assay and immunoblot were performed. Fluorescence microscopic and immunoblot assays were performed to confirm autophagy induction. The effects of RG were determined by evaluating body weight, tumor incidence, tumor volume and tumor burden in mice. Enzymatic and non-enzymatic antioxidant status was assessed. The role of some relevant signaling proteins was also analyzed. RESULTS： RG caused death ofA375 cells through induction of caspase 3-mediated apoptosis and Beclin-l-associated autophagy. Moreover, RG administration （75 mg/kg body weight） which showed no acute or chronic toxicity, showed significant reduction in the skin tumor burden of DMBA-painted mice. RG also demonstrated potent anti-lipid peroxidative and antioxidant functions during the course of skin cancer induction by DMBA. CONCLUSION： Chemopreventive potential of RG was demonstrated from overall results of this study indicating its possible use in therapeutic formulation of an effective drug to treat skin cancer.

Lymph node metastasis from non-melanoma skin cancer

The management of non-melanoma skin cancers metastatic to the neck is challenging due to variability in biological behavior and patterns of regional lymphatic spread.Metastatic non-melanoma skin cancers to the parotid and neck often behave aggressively,with a high incidence of local recurrence after treatment and reduced five-year survival outcomes.Patterns of lymphatic spread are different from those seen in mucosal squamous cell carcinoma,with higher prevalence of disease in the parotid and superficial lymphatics.These factors require that treatment is individualized to achieve optimal outcomes.Traditionally,the management of non-melanoma skin cancers metastatic to lymph nodes has involved surgical excision followed by adjuvant radiation therapy.However,novel systemic therapies are showing promising results and their role in the management of these cancers is evolving.

Genetic Predisposition to Numerous Large Ulcerating Basal Cell Carcinomas and Response to Immune Therapy

Objective:Well-defined germ-line mutations in thePTCH1 gene are associated with syndromic multiple basal cell carcinomas(BCCs).Here,we used whole exome sequencing(WES)to identify the role of patched-1 in patients with multiple,unusually large BCCs.Methods:A 72-year old patient presenting with numerous BCCs progressing to large ulcerating lesions was enrolled.WES was used to identify the pathogenic gene locus.Results:Genetic work-up by WES identified a homozygousPTCH1 nonsense mutation in the tumor tissue but not present in her blood cells or in non-lesional skin.In addition,heterozygous missense mutations were identified in three cancer-associated genes(EPHB2,RET,andGALNT12)in blood cells as well as in lesional and non-lesional skin.We also tested systemic immune therapy as a potentially beneficial approach to treat patients with numerous large BCCs on scatted areas of involvement.A rapid and sustained response to nivolumab was noted,suggesting that it is an efficacious drug for long-term therapeutic outcome.Conclusion:PTCH1,EPHB2,RET,andGALNT12 may potentially contribute to the synergistic oncogene driven malignant transformation manifesting as multiple,unusually large BCCs.

Effect of Q-switched alexandrite laser irradiation on dermal melanocytes of nevus of Ota

OBJECTIVE: To investigate the effect of Q-switched alexandrite laser irradiation on dermal melanocytes of nevus of Ota. METHODS: Multiple biopsies were carried out on 4 patients with nevus of Ota before and after laser irradiation. Altogether 11 samples were examined under light microscope and 14 under transmission electron microscope. RESULTS: Immediately after laser irradiation, the dermal melanocytes were destroyed, the melanosomes were degenerated with central vesicle formation within most of them, and intradermal round vacuoles appeared. The epidermis remained intact. Three months to 1 year after irradiation, the degenerated melanosomes and cell debris were scavenged mainly by macrophages. Dermal melanocytes gradually decreased. No fibrosis was found. CONCLUSION: Q-switched alexandrite laser can selectively destroy dermal melanocytes of nevus of Ota and treat the disease safely.