The Research Of The Relation Between NPY、miR-15a/miR-16 And Bcl-2 In Pituitary Adenomas

Objective: This research was designed to further study the relationship among NPY, miR-15a/miR-16, and Bcl-2 in nonfunctioning pituitary adenoma cells. And NPY, miR-15a/miR-16, Bcl-2 variation within 96 h, while observing non-functioning pituitary adenoma cells’ proliferation and cell cycle changes. To investigate the impact of development in non-functioning pituitary adenoma cells by the relationship of NPY, mi R-15a/miR-16, and Bcl-2. Further understand the role of NPY in the pituitary adenoma occurrence and development, and adding some new experimental data for the gene therapy of pituitary adenomas. Content: Each αT3 cells group was given different interventions, observed NPY, miR-15a/miR-16, Bcl-2 gene expression and protein levels, as well as cells in each group proliferative activity, corresponding changes in the cell cycle. Finally, for each set of data were statistically analyzed and compared to a conclusion. Methods:1. Cells Source: non-functioning pituitary adenoma cell lines(αT3 cell lines) provided by the Beijing Xiehe Medical College Hospital Neurosurgery Laboratory.2.Experimental groups: Ⅰ control group; Ⅱ NPY enhancer group; Ⅲ NPY antagonist; Ⅳ miR-15 a antisense nucleic acid molecules group; Ⅴ carrying miR-15 a plasmid group; Ⅵ mi R-16 antisense nucleic acid molecules group; Ⅶ carrying miR-16 plasmid group; Ⅷ NPY enhancer and miR-15 a intervention group; Ⅸ NPY enhancer and miR-16 intervention group.3. In each group, six indicators were applicated by RTFQ-PCR and Western Blot.4.Ⅰ, Ⅱ, Ⅲ, Ⅷ, Ⅸ group were detected by MTT, and flow cytometry. Results:1. After NPY enhancer/antagonist, NPY mRNA, NPY and Bcl-2 mRNA, Bcl-2 protein expression compared with the control group increased/decreased. NPY expression and miR-15a/miR-16 was negative.2. After the increase mi R-15a/miR-16, NPY mRNA, NPY, and Bcl-2 mRNA, Bcl-2 expression than the control group decreased or no significant change.3. After the cut miR-15a/miR-16, NPY mRNA, NPY, and Bcl-2 mRNA, Bcl-2 expression increased compared with the control group, or no significant change.4. While NPY enhancer and be raised miR-15a/miR-16, Bcl-2 mRNA and Bcl-2 expression is lower than when alone NPY enhancer alone is higher than the increase miR-15a/miR-16.5. NPY, Bcl-2 expression increased αT3 when mitotic cell cycle tends to change; NPY, while Bcl-2 expression decreased αT3 cells tend to cell cycle arrest.6. NPY expression Ⅱ group and αT3 cell proliferation significant positive linear correlation; Bcl-2 also αT3 cell proliferation positive relationship. Conclusion:1. Each group of NPY and Bcl-2 expression was similar to the trend, NPY and miR-15a/miR-16 expression was negative. Presumably by acting on miR-15a/miR-16 NPY may regulate the expression of Bcl-2 forward.2. Whole miR-15a/miR-16 and Bcl-2 mRNA, Bcl-2 expression was negative, negative effect on NPY mRNA and NPY miR-15a/miR-16 minor.3. Bcl-2 may be affected by both NPY and miR-15a/miR-16, more support NPY may influence miR-15a/miR-16 thereby altering the expression of Bcl-2.4. NPY and Bcl-2 may be associated with αT3 cell cycle and cell proliferation activity. Cell proliferation activity decreased after antagonize. Suggesting that antagonism of NPY might be one way of treatment of pituitary adenomas.5. Experimental results show that the role of Bcl-2 may be subject to inhibit the expression of NPY from being in the early mi R-15a/miR-16.6. Increased expression of NPY may affect pituitary adenomas, one of the factors of development; antagonize NPY expression but also for gene therapy of pituitary adenomas adds new experimental data, it is worth further study.