Black tea extract prevents inorganic arsenic induced uncontrolled proliferation, epithelial to mesenchymal transition and induction of metastatic properties in HaCaT keratinocytes - an in vitro study.

A major global problem is chronic exposure to inorganic arsenic (iAs) which causes various health hazards including cancer. Escalation of reactive oxygen species (ROS) generation by chronic iAs exposure promotes Epithelial to Mesenchymal transition (EMT) which is followed by metastatic progression. In the present study, skin keratinocyte cells (HaCaT) were divided into three groups: (i) untreated, (ii) chronically iAs exposed, (iii) black tea extract (BTE) along with iAs treated. ROS was estimated by flowcytometry, expression of EMT markers were assessed by flowcytometry, western-blotting and Immunofluorescence. For metastatic studies, wound-healing assay, gelatin zymography, western-blot, transwell migration/invasion assay had been performed. Long term exposure of HaCaT cells to iAs causes excess generation of ROS. Morphological transformation and EMT were apparent at 210 days of exposure. Development of metastatic characteristics were observed at 240 days. Alterations in the parameters induced by iAs were found to be ameliorated by BTE. BTE was found to quench excess generation of ROS by iAs, subsequently inhibiting the chain of events like EMT and metastasis. Therefore, BTE may be considered as a potential phytochemical to prevent the deleterious effect of iAs. Skin carcinogenesis induced by iAs may thus be prevented by BTE via inhibition of EMT.

Chemopreventive Role of Black Tea Extract in Swiss Albino Mice Exposed to Inorganic Arsenic.

OBJECTIVE: Chronic exposure to inorganic arsenic (iAs) may cause a number of health problems including skin cancer. Present study is aimed to look into the potential of black tea extract (BTE) to prevent the development of skin carcinoma in Swiss albino mice. METHODS: The study was done on Swiss albino mice, chronically exposed to inorganic arsenic. 150 mice were housed in different cages, 5 in each cage. The control mice did not receive any treatment. Mice were sacrificed at 30, 90, 180, 270 and 330 days. Development of carcinogenesis was assessed by histological studies. Generation of Reactive Oxygen Species (ROS) and Reactive Oxygen Species (RNS) were estimated using 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) and Greiss reagent respectively, and their consequences on DNA (by Micronuclei and Comet assay), protein (by protein carbonyl assay kit) and lipid (by lipid peroxidation) were estimated. Activity of antioxidant enzymes, along with total antioxidant capacity were measured by respective kits. Repair percentage was obtained by Comet assay. Western blotting was employed to study the expression of repair enzymes and expression of cytokines. Sandwich Enzyme-linked Immunosorbent Assay (ELISA) technique was employed to study the activity of various cytokines. RESULTS: At 330 days, invasive squamous cell carcinoma of the skin developed. With increasing time generation of ROS and RNS increased, causing damage to DNA, protein and lipid. Antioxidant defence system gets repressed with time. Capacity to repair the DNA damage is inhibited by iAs, due to alteration in repair enzymes - XRCC I, DNA Ligase I, PARP I, ERCC1, ERCC2, XPA, DNA Ligase IV, DNA PKc and Ku-70. Another consequence of iAs exposure is chronic inflammation due to disrupted cytokine level. Intervention with BTE reverses these deleterious effects, preventing development of skin carcinogenesis.