The Over-expression Of FGFR4 Could Influence The Features Of Gastric Cancer Cells And Inhibit The Efficacy Of PD173074 And 5- Fluorouracil Towards Gastric Cancer

Gastric cancer is one of the most common malignant tumor of the world, in Asia the incidence is higher than other continents. Morbidity and mortality is very high in our country. But, in our country, the discovery of early gastric cancer rate is about 20%, often in the terminal cancer or due to metastatic gastric cancer were found.So much for advanced gastric cancer in our country, and with poor prognosis, often need to give priority to with surgery, radiation and chemotherapy and other auxiliary treatment. At present, there are many adjuvant chemotherapy and palliative chemotherapy for gastric carcinoma, many of which platinum and fluorouracil for first-line drugs, such as cisplatin and fluorouracil, fluorouracil plus oxaliplatin into, DCF, ECF, etc., but its survival time is not ideal. In recent years, targeted drugs in the treatment of cancer of the stomach become a hot spot. Due to the occurrence of gastric cancer development is a multiple factors and multiple stages and multiple genes involved in the process of its pathogenesis is not clear, therefore, at present in addition to Hector plug butyl, most of the drugs did not make a breakthrough in the study of gastric cancer. So looking for new target genes and the occurrence of gastric cancer development mechanism has important effect to treating stomach cancer.Fibroblast growth factor(FGFRs) is a member of the immunoglobulin gene superfamily, distributed in a variety of cell membranes. In recent years, reports of FGFR family molecule in the different tumor growing, with FGFR4 polypeptide sex sites Gly388 Arg most studied. Some researchers believe that FGFR family has broad application prospects in diagnosis and treatment of malignant tumor. But as so far, FGFR4 research rarely in gastric cancer.FGFR4 in common malignant tumor such as pancreatic cancer, rhabdomyosarcoma, breast cancer and renal cell carcinoma were highly expressed. 20 ever found in a variety of fibroblast growth factor(FGF) in 10 can combine with FGFR4, only FGF19 combined with FGFR4 specificity.FGFR4 activated can make FGF19 promote liver cell proliferation, induction of liver cancer formation.Some reports suggest PD173074(PD) and FGFR family have very good affinity and selectivity, and PD can inhibit liver cancer cell proliferation. Some studies suggest that PD can reverse FGF19 specificity stimulation of AFP production increases, thus speculated that PD can inhibit the activity of FGFR4.The current FGFR4 less research in gastric cancer, so this topic discusses FGFR4 in its role in the development of cancer of the stomach, for the early diagnosis and prognosis of gastric cancer, providing strong theoretical basis for gene therapy.Through the body function in vitro study FGFR4 detection in different sources of biological characteristics of tumor cell lines express the similarities and differences, discuss fibroblast growth factor receptor 4(FGFR4) in gastric cancer(GC) function, and explore the agent for FGFR4 therapeutic value. By slow virus building FGFR4 RNAi, gastric carcinoma model used in nude mice, and in vivo in vitro correlation experiment, detect gastric cancer biology related protein expression and the changes. Using 5- Fu and PD173074 single-agent or joint, intervention slow virus and cell lines, the in vivo and in vitro experiments, discusses the joint after the two drugs on gastric cancer biological behavior and mechanisms of the influence of a more comprehensive study FGFR4 in its role in the development of cancer of the stomach, provide the basis for PD173074 applied in clinical treatment of gastric cancer, and provide a new evaluation index for the prognosis of gastric cancer.Part One The over-expression of FGFR4 could influence the features of gastric cancer cellsChapter One The over-expression of FGFR4 could influence the features of gastric cancer cellsBackground and projectiveGastric cancer is one of the most common malignant tumor of the world, in our country, the discovery of early gastric cancer rate is about 20%, often in the terminal cancer or due to metastatic gastric cancer were found. Many for advanced gastric cancer in our country, and the prognosis is poor, often need to give priority to with surgery, radiation and chemotherapy steadiness of comprehensive treatment. At present, adjuvant chemotherapy and palliative chemotherapy for gastric carcinoma, many of which platinum and fluorouracil for first-line drugs, such as cisplatin and fluorouracil, fluorouracil plus oxaliplatin into, DCF model, ECF, etc., but its survival time is not ideal. In recent years, targeted drugs in the treatment of cancer of the stomach become a hot spot. But in addition to Hector plug butyl, most drugs did not make a breakthrough in the study of gastric cancer. All looking for a new target genes, the occurrence of gastric cancer development mechanism has important effect to treating stomach cancer. Fibroblast growth factor(FGFRs) is a member of the immunoglobulin gene super family, distributed in a variety of cell membranes. In recent years, reports of FGFR family molecule in the different tumor growing, with FGFR4 polypeptide sex sites Gly388 Arg most studied. Some researchers believe that FGFR family has broad application prospects in diagnosis and treatment of malignant tumor. But so far, FGFR4 research rarely in gastric cancer. This study aims to slow virus carrier mediated, build into FGFR4 expression cell line, research on the gastric cancer cell line FGFR4 proliferation, invasion ability, the effect of apoptosis. Testing different treatment group effect the change of protein expression, discusses FGFR4 effect on gastric cancer cell biology behavior mechanism.MethodUsing the Real- time PCR detection FGFR4 HGC27 in gastric cancer cell line, MKN28, BGC823, SGC7901 and MKN45, expression of the 293 T, select lower expression of the cell. Build FGFR4 expression cell line, through CCK8 tested expression changes in cell lines and normal cell line proliferation. Flow cytometry sorting test FGFR4 influence on gastric cancer cell apoptosis, as well as by the changes of Transwell testing its ability to attack. Western Blot test different treatment group effect the change of protein expression.Result:This research build successful FGFR4 expression cell line, respectively set up normal cells, negative plasmid group and the expression of cell FGFR4 group. One of FGFR4 expression cells than normal group and negative plasmid in the control group and invasive ability are enhanced, hence reduce apoptosis rate. The expression of PCNA and Bcl – xl.ConclusionThis study screen out two strains FGFR4 cell lines in low expression from the various malignant degree of gastric cancer cell lines, and by lentiviral transfection to build it into FGFR4 high expression of gastric cancer cell lines. After by studying its expression, proliferation, invasion ability, apoptosis and related proteins, we found gastric cancer cell line proliferation strengthened after FGFR4 over expression, apoptosis rate reduced, invasive ability enhancement, presenting FGFR4 can promote gastric cancer cell growth, increase the malignant degree, promote gastric cancer cell invasive.Chapter Two The over-expression of FGFR4 could influence the features of Nude mice into gastric cancer tumorFGFR4 in common malignant tumor such as pancreatic cancer, rhabdomyosarcoma, breast cancer and renal cell carcinoma were highly expressed. 10 of more than 20 fibroblast growth factors(FGF) can combine with FGFR4, only FGF19 combined with FGFR4 specificity. Activated FGFR4 can make FGF19 promote liver cell proliferation, induction of liver cancer formation. The current FGFR4 has less research in gastric cancer, so this topic discusses FGFR4 in its role in the development of cancer of the stomach, for the early diagnosis, prognosis and gene therapy of gastric cancer providing strong theoretical basis.Injecting FGFR4 expression cell line and the corresponding untreatment of gastric cancer cell lines into the back of nude mice subcutaneously, and building nude mice model of gastric cancer. After 6-8 weeks inoculation, observing different volumes of tumors and the change of size indicators.Subcutaneous injection FGFR4 overexpress BGC803 and BGC823 cell lines in nude mice tumors had significantly greater volumes than the negative control group and blank control group, and the difference has statistical significance(P < 0.05).This difference is especially obvious in 10, 13 and 16 days days after cell injection.ConclusionOverexpression of FGFR4 can promote the growth of gastric cancer cells in the body. So we speculate that the high expression of FGFR4 protein may speed up the progress of the advanced gastric cancer.Part Two The over-expression of FGFR4 could influence the features of gastric cancer cells and inhibit the efficacy of PD173074 and 5-Fluorouracil towards gastric cancerBackground and projectivePD173074(PD) and FGFR family have very good affinity and selectivity.In recent years, some scholars think that PD can inhibit liver cancer cell proliferation. Although we can’t rule out PD may inhibit FGFR1, FGFR2 or FGFR3 to reach the purpose of inhibiting cell proliferation, but PD can reverse FGF19 specificity stimulation of AFP production increases, prompting PD inhibit cell proliferation mainly by inhibiting FGFR4.In addition, the PD for FGFR4 high expression of liver cancer cell lines Hu H7 d antiproliferative effect significantly better than FGFR4 lower expression of liver cancer cell lines and the cancer cells, thus further illustrate FGFR4, compared to the other FGFRs subtype of PD efficiency play a more important role.Little FGFR4 research in gastric cancer, so we will focus on FGFR4 in its role in the development of gastric cancer.Using PD173074 and 5-Fu treat FGFR4 expression cell line by single-agent or a combination of two drugs. respectively use no processing cell lines as a control, making a series of function test in vitro to evaluate the influence of the biological behavior of the gastric cancer cell lines with expression different level of FGFR4 by PD173074 and 5- Fu. At the same time, we detect the changes of cell proliferation, invasion, apoptosis and cell cycle related protein expression after different treatment factors processing at the molecular level.MethodUsing PD173074 and 5-Fu treat FGFR4 expression cell line by single-agent or a combination of two drugs. respectively use no processing cell lines as a control, making a series of function test in vitro to evaluate the influence of the biological behavior of the gastric cancer cell lines with expression different level of FGFR4 by PD173074 and 5- Fu. At the same time, we detect the changes of cell proliferation, invasion, apoptosis and cell cycle related protein expression after different treatment factors processing at the molecular level.Result:The gastric cancer cell line MGC803 and BGC823 cell proliferation was inhibited with 5- Fu or PD and combination of the two medicines increasing the rate of cell apoptosis. After constructed FGFR4 over expression cell lines by transfection technology, cell sensitivity of 5- Fu or PD and the combination of the two drugs decreased, cell proliferation inhibition of ease, apoptosis rate reduce comparing not the expression of cell lines.ConclusionThe gastric cancer cell line MGC803 and BGC823 cell proliferation was inhibited with 5- Fu or PD and combination of the two medicines increasing the rate of cell apoptosis. Over expression of FGFR4 may be able to reduce the effect of inhibition of gastric cancer cell line by 5- Fu and PD. If you want to get the same normal gastric cancer cells proliferation inhibition, you must increased 5- Fu and PD drug dosage to obtain satisfactory curative effect...