Effect of Standardized Decoction of <i>Nigella sativa</i> Seed, <i>Hemidesmus indicus</i> Root and <i>Smilax glabra Rhizome on the Expression of p53 and p21 Genes in Human Hepatoma Cells (HepG2) and Mouse Liver with Chemically-Induced Hepatocarcinogenesis

Abstract

Purpose: To evaluate in vitro (using human hepatoma HepG2 cells) and in vivo (using mouse liver with diethlynitrosamine (DEN)-induced hepatocarcinogenesis) effect of a standardized decoction on the expression of p53 (tumour suppressor) and p21 (cyclin kinase inhibitor) genes with the long-term goal of developing the formulation into a globally acceptable therapy for hepatocellular carcinoma (HCC). Methods: The effect of the decoction on (a) mRNA and (b) protein expression of p53 and p21 genes in HepG2 cells and mouse livers with DEN-induced early hepatocarcinogenesis were evaluated by (a) reverse transcription PCR (RT-PCR) and (b) immunohistochemical and Western blot analysis, respectively. Results:The results demonstrated that the decoction significantly (p < 0.001) enhanced the expression of p53 and p21 genes in a time-and dose-dependent manner in HepG2 cells. A dose of 75 µg/ml significantly increased p53 mRNA at 24 and 48 h and p21 mRNA at 12, 24, 48 h of incubation with the decoction (p < 0.01). Induction of hepatocarcinogenesis in mice significantly increased hepatic expression of both p53 and p21 compared to distilled water control (p < 0.001), while treatment with the decoction further enhanced expression of both genes in DEN-induced hepatocarcinogenesis (p < 0.01). Conclusion:Overall, the findings demonstrate that the decoction may mediate its reported antihepatocarcinogenic effect, at least in part, through the modulating activities of genes involved in tumour suppression and cell cycle arrest.

Modulation of apoptosis in human hepatocellular carcinoma (HepG2 cells) by a standardized herbal decoction of Nigella sativa seeds, Hemidesmus indicus roots and Smilax glabra rhizomes with anti- hepatocarcinogenic effects

“…Previous in vivo and in vitro investigations [ 1 , 2 ] have shown that the decoction, comprised of N. sativa seed, H. indicus root, and S. glabra , can mediate anti-hepatocarcinogenic effects, without producing any significant toxic side effects. The antihepatocarcinogenic effects of the decoction probably results from an impact of this polyherbal formulation on a broad range of biochemical activities including antioxidant activity [ 5 ], anti-inflammatory activity, immunomodulatory activity and modulation of activities of factors involved in cell proliferation and cell cycle arrest [ 3 , 4 , 6 ]. In the study carried out by Thabrew et al , (2005), a flow cytometric analysis of HepG2 cells exposed to the test decoction indicated that this polyherbal formulation also has the potential to induce apoptosis.…”

“…The decoction has been standardized considering its High Performance Liquid Chromatography (HPLC) profile and physico-chemical parameters according to WHO guidelines [ 3 ]. Further in vivo and in vitro investigations have demonstrated that cytotoxicity, [ 3 , 4 ] antioxidant activity, anti-inflammatory activity [ 5 ] and up regulation of p53 and p21 activities [ 6 ] are some of the possible mechanisms through which the above decoction may mediate its anti-hepatocarcinogenic action.…”

Dual Regulating Effect of Shaoyao-Gangcao-Tang on COX- 2 Expression in Acute and Resolution Phases of Carrageenin-Induced Pleurisy in Rats

“…Inflammatory cell pellets were lysed in RIPA buffer ( 50 mM Tris-HCl (pH 7.6), 150 mM NaCl, 1 % NP-40, 0.5 % sodium deoxycholate, 0.1 % SDS) [17] and EDTA-free protease inhibitor cocktail (Roche, Basel, Switzerland). Equal amounts of protein samples were separated by SDS-PAGE electrophoresis and transferred onto PVDF membranes (Milipore, MA, USA).…”

Promotion of HepG2 cell apoptosis by flower of Allium atroviolaceum and the mechanism of action

“…There is no beneficial cure for this malignancy [ 10 ] and the recovery rate of HCC is low in most cases [ 11 ]. In addition, the inevitable side effects, such as toxicity to normal cells and bone marrow illustrate an instant demand to search for better methods and novel anti-cancer agents that would decrease the mortality rate of HCC with fewer side effects [ 12 ].…”