| The prostate cancer always in elder man, The therapeutic meathod depends on androgen that surgical operation, radiotherapy, and the castration operation etc. in the past. There are seldom clinical symptom In primary prostate cancer, so it is detected late or after metastasis. Up to now ,we have no effective treatment for the advance prostate cancer. However, the gene therapy became new therapies can inhibit tumor cell differentiation and induce apoptosis. The result shows, the overexpress GRIM-19 gene can inhabit the prostate cance cell PC-3M differerntiation and induce the apotosis .Consquently it will therapy prostate cancer the day afert tomorrow.The GRIM-19 gene is a new gene which induce express of apotosis regulatory factor. Overexpress GRIM-19 can increase IFN/RA induce mammary cancer cell apotosis. The GRIM-19 is similar as many kinds of tumor cell inhibitory factors, as P53 P300/cbp and RB gene. But it is not any known apotosis factor which express majority tissue generally. GRIM-19 is a nucleus protein that locate in chromatosome 19P13.1. There is the indispensable domain which can inhabit prostate caner. It will be a new tumor inhabit factor.Objective:In this experiment, we choose human prostate cancer cell PC-3M as the study object. We approach the inhibition of GRIM-19 to the generation of the human prostate cancer cell and its probable mechanism through in viro experiments. It will provide a new way for the treatment of prostate cancer.Methods:1. Immunohistochemistry stain to detected expression of GRIM-19 in human prostate tissue 2. To succeed construct pGRIM-19 recombinant plasmid.; 3. The inhibition effect of pGRIM-19 to the PC-3M cell were detected by MTT experiment 4.Use acridine orange to detect the cell apoptosis; 5. to detect the cell cycle by FCM;6.Western blot detected the GRIM-19 in tranfection PC-3M cells 7. The expression of GRIM-19 mRNA in tumor tissue by RT-PCR;Results:1. Immunocytochemistry stain showed that the expression level of GRIM-19 protein was express in normal prostate tissue lower express in cancer tissue.2. Recombinant plasmid was transfected into PC-3M cells by liposome, and transfected PC-3M cells successfully. The transfected cells were screened by G418. We acquired stably transfected PC-3M cells with pcDNA3.1 empty plasmid, called empty plasmid group, and pcDNA3.1 recombinant plasmid with GRIM-19, called GRIM-19 group. Assessed thehigh expression level of GRIM-19 in PC-3M cells by RT-PCR andWestern blot.3. The mRNA expression of GRIM-19 groups are higher than control group.4 Western blot shows the GRIM-19 group protein express higher then other two groups.5. Grow curve and MTT chromatometry indicated that theproliferation ability of TIMP-3 group cells were significantly lower than those of untransfected group and empty plasmid group(p<0.01). The inhibition ratio of PC-3M cell growth was 48.3%.6.The result of flow cytometry (FCM) shows, The pGRIM-19 group found aptosis cells.Apotosis rate is 24.4%.7. Acridine orange detected the apotosis cell .8.The VEGF,Bcl-2, c-myc ,cyclin D1 apotosis factor is down regulate by GRIM-19.Conclusions:1) The expression of GRIM-19 is inhibited in prostate cancer tissue, the expression of GRIM-19 is negative correlation with malignant tumor.2) pGRIM-19 could inhibit proliferation of the human prostate cancer cell PC-3M in vitro.3) pGRIM-19 could induce human prostate cancer cell PC-3M with apopotsis.4) pGRIM-19 induce apotosis correlate with Bcl-2, c-myc, cyclinD1, VEGF express regulation...
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