| Objective:In China, hepatocellular carcinoma (HCC or liver cancer) has risen for the second place. Every year, 130,000 patients die of liver cancer, accounting for 43% of total worldwide liver cancer deaths. Traditional and new minimally invasive surgical treatment of surgical resection is the main treatment. However, the tumors are too large and poor location, poor liver function reserve or portal hypertension often limit the indications for surgical treatment. and the high relapse rate after surgery is often successful wasted. Liver transplantation can not be universal, because of a shortage of donor organs, even if it is early cancer, the beneficiaries are very limited. HCC is not sensitive to chemotherapy or easy Lai properties. TACE is not extended as the only possible survival of patients with unresectable liver cancer treatment. Therefore, the majority of HCC patients can not provide any effective treatment. It can be said that in cancer, liver cancer is more need for a new treatment. Tumor necrosis factor-related apoptosis-inducing ligand (TNFrelated apop tosis induci ng ligand TRA IL) found that To develop a new generation of high efficiency and low toxicity, tumor selective display of a good prospect. TRAIL can be killed with two receptors (DR4 and DR5) interaction. selectively induce apoptosis in cancer cell lines organization sources; and decoy receptors (68.8. ovarian cancers) to complete the intracellular death domain, the signal can transmit apoptosis. it inhibits TRAIL-induced apoptosis. As protects normal cells and induced apoptosis characteristics of a source of widespread concern. In recent years, some form of further study found that TRAIL can induce apoptosis in normal liver. Therefore TRAIL recent clinical application of the safety advantages of a anxiety. Activation of DR5 developed antibodies to stimulate tumor cell apoptosis signal to the anti-tumor therapy is another effective way Therefore, we used anti-human DR5 single alternative TRAIL antibody for the treatment of tumors. In this study, we detected expression of DR5 and whether SMMC7721 cell surface expression. mRNA and protein levels, mainly through confirmation. The application of anti-human DR5 monoclonal antibody SMMC7721 induced apoptosis; Research on anti-human DR5 monoclonal antibody with 5-fluorouracil and cisplatin. SMMC7-7721 adriamycin-induced apoptosis of synergies. To reduce the dose of drugs, reduce side effects and increase the therapeutic effects. Biological treatment of hepatocellular carcinoma experimental basis.Methods: (1) Conventional SMMC7721 liver cancer cell lines. DR5 application-specific RT-PCR primers do experiments SMMC7721 cells confirmed hepatocellular carcinoma cell lines have DR5 mRNA expression DR5 detected using flow cytometry in human hepatoma cell line SMMC7721 of expression.(2) were determined by flow cytometry and MTT different concentrations of anti-human DR5 monoclonal antibody induced SMMC7721 Fine apoptosis; the same time for different concentrations of anti-human DR5 monoclonal antibody SMMC7721 induced apoptosis. anti-human DR5 monoclonal antibody with 5-fluorouracil and cisplatin. perspective of experimental results. Apoptosis induced by adriamycin SMMC7-7721 synergies, DNA fragments of cell apoptosis detection kit; Saddam Kyi was fine, morphology of apoptosis was observed from the perspective of experimental results.Results: (1)DR5 mRNA expression:β-actin and DR5 with reference to the RT-PCR product by agarose gel electrophoresis. The results showed that the liver cells SMMC7721 DR5 mRNA expression; SMMC7721 cells by flow cytometry for cell DR5 percentage of positive cells were detected DR5 from 89.0%to94.8%, with an average of 91.2%. (2)With different concentrations of anti-human DR5 monoclonal antibody (0.625μg/ml, 1.25μg/ml, 2.5μg/ml, 5μg/ml, 10μg/ml, 20μg/ml, 40μg/ml, 80μg/ml ) and20μg/ml of anti-human DR5 monoclonal antibody was with doxorubicin 5μg/ml. 5μg/ml 5-FU, in combination with cisplatin 2.5μg/ ml hepatoma cell line SMMC7721. MTT and the growth curve showed that anti-human DR5 monoclonal antibody inhibited the growth of liver cancer cell line was SMMC7721 time-and dose-dependent manner. anti-human DR5 monoclonal antibody in combination with low concentrations of chemotherapeutic drugs SMMC7721 cells than single anti-human DR5 polyclonal antibody or low concentrations of chemotherapeutic drugs SMMC7721 single role in the inhibition of cell growth more evident. Flow cytometry showed different concentrations of anti-human DR5 monoclonal antibody role in SMMC7721 cells. With increasing concentration of apoptosis increased, and low concentrations of anti-human DR5 monoclonal antibody chemotherapy (doxorubicin, 5-FU. CDDP) in the role of SMMC7721 cells than lower concentrations of anti-human DR5 monoclonal antibody or chemotherapy (Epirubicin-, 5-FU, cisplatin), the role of a single cell apoptosis significantly increased, the difference was significant. SMMC7721 anti-human DR5 monoclonal antibodies can induce apoptosis; anti-human DR5 monoclonal antibody with 5-fluorouracil and cisplatin. ADM SMMC7-7721 synergistically induced apoptosis in hepatoma cell line. (3)DNA ladder Frederick Side apoptosis appeared180-200bp DNA fragments In gel electrophoresis showed VSMCs map (DNA ladder); Ji Saddam small size smaller staining of apoptotic cells, the cytoplasmic concentration of nuclear chromatin highly entwine. Many of the phenomenon called cavitation cavity; gather the nuclear chromatin, marginalization; Apoptosis in late nucleus split to pieces, produced apoptotic bodies.Conclusion : (1)Of DR5 hepatoma cell line SMMC7721 cell surface expression. (2)In vitro studies have shown different concentrations of anti-human DR5 monoclonal antibody against hepatocellular carcinoma cell lines SMMC7721 Health long time-and dose-dependent inhibition in drug concentration was 2.50 ml, the rate of inhibition 20.49%; But the concentration was 20 ml, the inhibitory rate of 52.85%. Both significant difference (P <0.01) from the curve would appear that the larger the slope of the line was a linear relationship. When the concentration was from 20 to 80μg ml ml, the relative inhibitory rate of increase is slow, However, to show growth in a dose-dependent linear relationship. These results show that anti-human DR5 monoclonal antibody-induced apoptosis in a dose-dependent SMMC7721. But there are also the best drug-dependent concentration of 2.5μg to 20 ml, respectively.(3)anti-human DR5 monoclonal antibody with doxorubicin, 5-fluorouracil and cisplatin combination. SMMC7721 the inhibition rate was higher than the chemotherapy alone group (P <0.01). In addition, the combination drug group on SMMC7721 cells was significantly inhibited by anti-human DR5 monoclonal antibody group than alone. with anti-human DR5 monoclonal antibody combined with the effects of adriamycin most notable. The results showed that the increase of anti-human DR5 monoclonal antibody with chemotherapy-induced apoptosis of SMMC7721. DR5 monoclonal antibody against human hepatoma cell line SMMC7721 induce apoptosis of cells, The biological therapy for hepatocellular carcinoma experimental basis. Anti-human DR5 monoclonal antibody combined with low-dose chemotherapy alone could improve the anti-human DR5 induction. alone can enhance the effect of low-dose chemotherapy drugs, we will be able to appropriately reduce the dose of drugs, reduce toxicity and increase efficacy for new ideas. |
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