الفهرس 
1. Introduction and Aim of the WorkOnly 14 pages are availabe for public view
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Abstract
In the present work, we studied the effect of a combined therapy of the commercially available product of a cocktail of seven phytochemicals ””””BSG”””” along with chemotherapy aiming to increase the chemosensitivity of HepG2 cell line to cancer chemoprevention Docetaxel drug. Moreover, the molecular mechanisms mediating the combined therapy inhibitory effect on HepG2 cell proliferation, survival and migration was studied as well. In current work, we used the DPPH assay and determination of flavonoid, polyphenols, and tannins contend for BSG characterization, the data showed that BSG extract contains (18.1025 mg Catechin equivalent/gm BSG, 109.365 mg gallic acid equivalent/gm BSG and 30.944 mg gallic acid equivalent/gm BSG) compared to catechin and gallic acid as reference standards, respectively. In current study we used HepG2 cell line as an in vitro experimental model of liver cancer cells. The effect of BSG on cell proliferation and the effective dosage was determined using cell proliferation assay. The cells were cultured in the presence of ascending doses of BSG (5000 μg/ml, 2500 μg/ml, 1250 μg/ml, 625 μg/ml, 312.5 μg/ml, 156.25 μg/ml, 78.125 μg/ml μg/ml, 39.06 μg/ml, 19.53 μg/ml and 9.76 μg/ml). Measurements of the absorbance of the reduced form of MTT indicated a significant effect of BSG on diminishing cell proliferation and the half inhibitory concentration (IC50) was determined to be (263 μg/ml). Relying on these results, molecular studies of the effects of BSG and/or DOC on HepG2 cells were conducted by sub-culturing the cells into six groups as follows; • group 1: HepG2 control • group 2: HepG2 vehicle control • group 3: HepG2 treated with IC50 of BSG (263 μg/ml) • group 4: HepG2 treated with of BSG (131 μg/ml) • group 5: HepG2 treated with of BSG (131 μg/ml) + Docetaxel (0.125 μg/ml) • group 6: HepG2 treated with Docetaxel (0.125 μg/ml). The data of BSG and/or Docetaxel on cell proliferation was confirmed by PCNA using immunocytochemistry techniQue The effect of BSG and/or Docetaxel was compared with the untreated group (control) and with the vehicle control which contains the solvent of drugs under study. On the other hand, the effect of treatments on HepG2 cell motility, as an essential step in cancer cell metastasis, was conducted using wound healing assay. The resulted data revealed a significant decrease at the cell ability to migrate into the generated scratch in groups treated with combination between BSG with DOC, DOC and BSG, respectively. The data was confirmed by measuring the CD44 expression by convential PCR as a marker of cell migration. In order to elucidate the effect of BSG and/or DOC on cell proliferation, the change at phases of cell cycle of all studied groups was analyzed using immunodetection. The data indicated a downregulation at the p21 as downstream targeted p53 upon treatment with BSG, DOC and the combination, respectively. These results were further confirmed via immunodetection of cell pro-apoptotic gene Bax using protein ELISA assay. The resulted data revealed a significant redundant reduction at Bax expression in cells of BSG IC50 (0.263 mg/ml), BSG (0.131 mg/ml), DOC(0.125 μg/ml) and their combination BSG (0.131 mg/ml) with DOC( 0.125 μg/ml), respectively. Moreover, this study illustrated the expression of the antiapoptotic proteins BCL2 and SVV Groups treated with BSG (0.131 mg/ml), DOC(0.125μg/ml) and combination BSG (0.131 mg/ml) with DOC (0.125 μg/ml), respectively. The effect of BSG and/or DOC on drug resistance by using convential PCR of MDR1 expression didn’t show any significance. In our study the treatment with BSG and/or DOC upregulating the expression of LC3II which is a marker for autophagy. On the other hand, the treatment BSG IC50 (0.263 mg/ml) downregulated the expression of Nanog. This data indicated the possible implication of the treatment in cancer cell stemness and autophagy which needs a further prospective assessment such as P62 which is a copartner of LC3II.