Expression Of PTTG In Human Glioma And Construction Of SiRNA Targeting PTTG

Objective: To construct silent and high efficient PTTG in view of the human brain sol lump cell line U251 the RNAi carrier, and transfect the U251 cell, and discuss its impact on PTTG gene expression in order to seek new target to treat the sol lump.Methods: Examine 72 examples of normal tissue of sol lump patients and the expression situation of PTTG protein of sol lump tissue; obtain cDNA (NM:004219) sequence of PTTG from Genebank; according to the Ambion Corporation online design software, screen three target sequences with their lengths respectively being 19 bp, designs these three pairs of nucleotide sequence to direct the thing, synthesize the small interference RNA fragment of PTTG specificity interference and link them with the pGenesil2 carrier to successfully constructs the PTTG interference carrier (pGenesil2-PTTG siRNA). Transfect pGenesil2-PTTG siRNA into U251 cell using the lipid .After 48 hours, collect cell to determine efficiency of transfection and observe conditions of transfection by using stream mode cell device and fluorescence microscope respectively. Using stream mode cell device and Western blot method to measure the PTTG protein expression conditions in each group.Results:1. PTTG is widely expressed in human brain sol lump, and its expression intensity increases along with the tumor malignant degree.2. Three reorganization carriers construct successfully.3. After transfection of U-251 cell, three pairs of recombined particle carrier can reduce the level of the sol lump PTTG protein expression quantity and the mRNA level obviously.Conclusions: PTTG in sol lump cell is widely expressed, especially in high rank tumor. The sol lump U251 cell is introduced the PTTGsiRNA gene to obviously reduce PTTGmRNA and the protein expression, thus suppressing the U251 cell multiplication, and suppressing the tumor the formation. This discovery proposes new content and direction for the sol lump's treatment.