Right after 48 h, complete RNA was extracted for Bcl 2 mRNA expression utilizin
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Right after 48 h, complete RNA was extracted for Bcl 2 mRNA expression utilizin
Discussion Convincing proof from each animal experiments and professional clinical scientific studies suggest that CpG ODN alone or in combination with other therapeutics are practical to the therapy of malignant tumors, From the current experiment, we first of all demonstrated that CpG ODN could appreciably improve the chemosensitivity of 5 FU in HepG2 human hepatoma cells in vitro. The mechan ism was related to CpG ODN mediated the expression of anti apoptotic aspects inside tumor cells, then indu cing apoptosis and cell cycle arrest at S phase. These findings supply new understanding of CpG ODN mediated direct cytotoxic results and new insights to the application of chemosensitizer. Presently, a number of clinical trials and animal ex periments have confirmed the activation of several immune cells and also the manufacturing of cytokines induced by CpG ODN have sizeable impact of anti tumor, CpG ODN is thought of like a likely chemo sensitizer with unclear mechanism. Former studies re ported that Peritumoral CpG ODN1826 therapy induces modulation of gene involved in DNA repair and sensitizes cancers cells to DNA damaging cisplatin treat ment in human IGROV 1 ovarian tumor cells, and peritumoral injection of CpG ODN1826 in blend with subcutaneous injection of 5 FU inhibit tumor growth and reverse the immunosuppression brought about by the therapy 5 fluorouracil in murine hepatoma, Even so, the direct cytotoxicity toward HepG2 cells will not be investigated in vitro. In our experiment, the results demonstrated that CpG ODN could significantly in crease the chemosensitivity of 5 FU in human hepatoma HepG2 cells in vitro. Apoptosis was imagined to become the major reason of cell death induced by chemosensitizer. Current studies indicated that stimulation of TLR9 with CpG ODN enhanced apoptosis in murine or human tumor cells, but ODN M362 promotes cell proliferation and survival in hu guy hepatocellular carcinomas, So we evaluate the direct cytotoxicity of CpG ODN and ODN M362 toward HepG2 cells, our success showed that CpG ODN induced significant inhibition of your survival of HepG2 cells, and ODN M362 had not direct cytotoxicity toward HepG2 cells, we upcoming documented that apoptosis was respon sible for CpG ODN and or 5 FU induced cytotoxicity of HepG2 cells applying MTS assay, observation of cell morph ology, Hoechst 33258 staining, and annexin V FITC staining, these final results indicated that CpG ODN increased the chemosensitivity of 5 FU in HepG2 cells by expanding apoptosis without the need of the have to have for immune system of host. Although many studies had been fo cused about the immunotherapeutic applications of CpG ODN by modulating immune technique of tumor bearing hosts, some current data showed that direct cytotox icity towards tumor cells is promising for treatment of various malignancies, These former scientific studies strongly sup ported our research, these outcomes showed that CpG ODN immediately induced apoptosis and enhanced the chemosensi tivity of 5 FU in HepG2 human hepatoma cells. Cell cycle arrest was believed to get another important rea son of cell death induced by anti tumor medicines, Fluorouracil is actually a pyrimidine analogue which is transformed inside the cell into various cytotoxic me tabolites and then incorporates into DNA and RNA, fi nally inducing cell cycle arrest and apoptosis by inhibiting the cells capacity to synthesize DNA.
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