Biological Effect Of Galectin-in Invasive Mechanism Of Pituitary Adenoma

1.To observe the expression of Galectin-3 in human pituitary adenomas2.Galectin-3 gene silencing was appliaed by RNAi3.To set up the cultured system of human pituitary adenoma lines4. hoping to evaluate its possible role in the mechanism of pituitary adenomas'invasion. To find a possible clinical marker of invasion. And to find a possible target of treatment of invasive adenomas.1 The expression of Galectin-3 in the human pituitary adenomas was detected by immunohistochemistry, and Galectin-3 is whose protein expression more strongly level was compared between patients of invasive and non-invasive adenoma;Galectin-3 is whose mRNA expression more strongly in invasion pituitary adenoma or noninvasion pituitary adenoma was detected by CISH.2 A high specific delivery system for siRNAs mediated by retrovirus vector was established. By this system, siRNAs can be integrated into the cell genome stably.We design and synthesized the shDNA, which can transcribe into shRNA and interfere with Galectin-3 gene. The shDNA duplex was ligated into the recombinant vector pGCL-GFP/U6 plasmid.the positive colonies of pGCL-GFP/U6 Gal-3shDNA-lwere selected by double restriction enzyme digestion with Hpa I and Xho I, Cell line was transfect with pGCL-GFP/U6 Gal-3shDNA-1 using pGCL-GFP/U6-s crambled shDNA as control. The expression level of Galectin-3 was detected by RT-PCR and Western Blot; then PI flow cytometric analysis were performed for apoptosis, MTT assay for cell proliferation.3 The prolactin adenoma derived cell-F cultured in vitro was in good condition and maintained the ability of proliferation and differentiation.1. The expression of Galectin-3 in the human pituitary prolactin adenomas and pituitary Adrenocorticotrophic hormone adenoma.2. In invasive adenomas, the Galectin-3 had a higher expression level than While in non-invasive adenomas, the difference was not so obvious.3. After when high the expression of Galectin-3-protein was detected by IHC method, CISH was used to detect the expression of Galectin-3-mRNA.4 A lentivirus RNAi vector containing shRNA targeting Galectin-3 gene was successfully constructed and be in efftct targeting Galectin-3 gene and will be used as method to develop Galectin-3 gene silencing therapeutics.5. The recombinant vector were successful constructed which was confirm by sequencing.The in vitro experiment indicated that the expression level of Galectin-3 were down regulated by western blot and RT-PCR analyses than control group (P<0.05); The interfering efficiency of Lentiviral was 95%, MTT assay showed that the proliferation of MCF-7 were suppress to(0.40±3.69)%than control group (0.71±0.156)%.FCM assay indicated that Lentiviral vector plasmid induce apoptosis of the cell about 68.78%.6 The cultured system of human prolactin pituitary adenoma derived cell-F was set up1. Galectin-3 might as a New molectin phyology marker of operation later invasion action of pituitary adenoma by judge2. Galectin-3 can judged phyology action of invasion prolactin pituitary adenoma3. Galectin-3 might have change in invasion pituitary adenoma was detected by chromogenic in situ hybridiza tion (CISH).4. A lentivirus RNAi vector containing shRNA targeting Galectin-3 gene was successfully constructed will be used as method to develop Galectin-3 gene silencing therapeutics.5. RNA interference can successful interfere with the expression of Galectin-3 in culture tumor cell and may affect the occurrence and development of the tumor.6. The cultured system of human prolactin pituitary adenoma derived cell-F have been set up.