| Recently, the incidence of malignant tumors showed an upward trend in the world. At present, 10 million new cancer patients are being increased all over the world every year. If we took effective preventive measures, the annual incidence rate will be reduced to 3 / 4 of the rate nowadays as is expected. Data in our country showed that 900,000 new cancer patients have been increased every year in our country since the 1970's. 1.3 million people have died of cancer each year. Cancer has become the first killer of mankind Whether international or domestic. Scientists are constantly exploring new methods to cure cancer. At present, the main methods including surgery, chemotherapy,radiotherapy,biological,are unsatisfactory. Medi- cat- ion therapy in malignant tumors of the three played an important role and developed the fastest.The development of anti-cancer drug, including genetic engineering, computer simulation and design of biomimetic natural anti-cancer activity is based on components. In addition, the application of genetically engineered bacteria producing special medicinal ingredients such as active protein or peptide drugs, are key point of biological medicine of our country and the world and is also the development of biotechnology industries and basic medicine. We know there are many kinds of protein having biological activity found in a variety of living organisms. That play a role in immune regulation in the experimental animal or human body. Purification of the activity of living organisms in the protein composition, with a view to finding some new clinical application of a protein or peptide, A large number of researches of domestic and abroad have become an important direction of work.From the end of 80's in the 20th century, the pharmacology of fungal immunomodulatory protein was deeply investigated in domestic and abroad . First of all, An active protein --8 (LZ-8)was isolated from Ganoderma lucidum hyphostroma . relative molecular weight of 12.7 KD, gene-coding region have about 330-350 base pairs, the protein product about 110-114 amino acids,acetylation blocks N-side.LZ-8 could inhibit the anaphylaxis, aggregate the action of red blood cell , excite peripheral lymphocyte of human proliferation and IL-2, TNF-αand other cytokines,immunological activity and so on.However, The investigation of the anti- S180 ascites tumor has not been reported. In this paper, the experiment in vitroexplored the immunomodulatory recombinant protein of Ganoderma lucidum (rLZ-8) anti-tumor mechanism.Expeiment in vivo:The selection of mouse ascites liver tumor cell the S180: domestic researchers investigated deeply S180 for immunology of biology,cell lines and set up the experimental conditions, transplanted to do more in depth research. Because the model has advantages ofshort time, high success rate against the characteristics of good homogen- eity, the current of this model has been widely used in tumor research and a tumor model recognized. In this study, the first rLZ-8 on S180 in vivo anti-tumor test, after the recovery of the S180 cells, passage and right hind hip subcutaneous inoculation, the success rate of model is 100%. This model has obvious external features, that it can be observed easily to, measure the growth of subcutaneous tumor size in order to reflect the efficacy of the experimental group.The comparative analysis of results in the solid tumor weight and growth rate showed that: With the increase in the concentration of rLZ-8 up to a certain extent, the tumor growth become lower and the tumor weight lighter. Although the treatment group of rLZ-8 and the inhibition rate was still inadequate compared to the control group, but rLZ-8--high, medium and low dose group, respectively, the inhibitory rate reached 43.6%, 28.7%, 19.8%; by comparing the statistical treatmen with normal control group .there was a significant difference (P <0.05), and it suggests rLZ-8 on S180 has a certain role in anti-tumor activity.From the comparative analysis of the mouse's weight and weight change in the extent of the growth curve experiment in each group, the average weight of rLZ-8-low, medium and high dose group is 34.2g, 36.1g, 37.9g. It is found that with the increase of rLZ-8 concentration the average weight of mice decreased.To some extent, rLZ-8 could inhibit the growth of ascites tumor cells S180. rLZ-8 anti-tumor effect of the treatment group significantly, and the normal control group, negative control group and positive control group showed significant differences (P<0.05).Experiment in vitro:rLZ-8 on the role of anti-S180 in vitro experiments is based on observation of CPE and MTT cell viability determination, which calculated the experimental drugs on the toxicity of the concentration of S180 cells.the results can be seen from the rLZ-8 in experimental group compared with normal control group, there is significant difference, indicating that rLZ-8 cells in vitro on the S180 has a strong role in the destruction.The rLZ-8 induced by NK, LAK cell cytotoxicity experiment. Drug group rLZ-8 --- a high concentration of low NK cell killing activity as follows: 16.9%, 28.3%, 32.5%, 38.6%, 56.9%. drug group rLZ-8—a high concentration of low-LAK cell killing activity as follows: 20.7%, 25.1%, 33.6%, 40.8%, 53.4%. As can be seen, rLZ-8 concentration of the higher of the two kinds of NK cells also increased.Apoptosis was detected by flow cytometry experiments, in strict accordance with the AnnexinV/PI, PI apoptosis detection kit instructions, and pay attention to each order of reagents added. From Annexin V/PI double staining graphs, charts can be seen, rLZ-8 drug groups were found to apoptotic cells, by different concentrations of drug treatment rLZ-8, the low, middle and high apoptosis rate of the drug group was 16.2%, 25.4%, 42.1% higher than the normal control group, indicating the rate of S180 -induced apoptosis rLZ-8 with the increase of the concentration has increased. Annexin V/PI detection of apoptosis are currently the best way. PI staining in a single map, table saw, rLZ-8 drug groups were apoptotic peak appeared, with different concentrations of drug treatment rLZ-8, the apoptosis rate of each group were 11.3%, 26.5%, 31.7%, The results showed that treatment with increased concentration, S180 cells also enhanced the apoptotic rate trend, which is particularly high concentration group, apoptosis rate of 31.7 percent. This note rLZ-8 caused apoptosis S180 is a very important role in anti-tumor mechanisms. However, its mechanism needs further study and we hope that it can be resolved in future experiment.The investigation suggests this rLZ-8 has a significant anti-tumor effect. So far, it has not been reported yet that the immunomodulatory regulatory protein conducts clinical trials of anti-tumor,and the fungal immunomodulatory protein has a wider development space and a greater excavation potential. With the depth of its research, it is expected to become an effective tumor biological drug in clinical treatment. |
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