siRNA interfere with growth and invasion of U251 cell by reducing expression of Moesin
AIM: To investigate the inhibitory effect of RNA interference ( RNAi) on the membrane structure extending spike protein ( Moesin) gene expression in human brain glioblastoma (U251) cells and the influence of Moesin to the proliferation and invasion capacity of the U251 cells. METHODS; In human glioma cell strain U251 as the research object, design small interfering RNA fragments for Moesin encoding gene, transfection to U251 cells, use reverse transcription polymerase chain reaction ( RT-PCR) to screen of efficient siRNA silencing fragment, and use immunoblotting( Western Blot) to detect the expression level of protein Moeisn after transfection. Tetra methyl-Thiazolyl-blue was used to assay the ability of proliferation, flow cytometry was performed to detect cell apoptosis, Transwell was performed to assay invasion changes, and scanning electron microscopy was applied to observe cell morphology changes after transfection. RESULTS; RT-PCR and Western Blot screen of a MOESIN-139 fragment of the best silence. siRNA transfected U251 cell growth was slowed down, in the 24-48 h period, siRNA group of U251 cells growth rate significantly lower than the blank group and the negative control group; and increased apoptosis, siRNA interference average apoptosis rate (17. 30 ± 2.01)% , was much higher than the blank group in average apoptosis rate( 1. 95 ±0. 33)% , negative group the average apoptosis rate of(2.02 ±0. 28) %. After siRNA transfection, the number of U251 cells went through the cell membrane Polycarbosilane ester significantly reduced from the blank group 26.47 ±4.07 and negative group 24.27 ±3. 63 reduced to the transfection group 11. 53 ±2. 61. Changes in cell morphology, cell surface pseudopodia decreased from blank group 14. 2 ± 2. 58, negative group 15. 8 ±1. 30 to transfection group of 6.6 ± 1. 82 ( P < 0. 05 ). CONCLUSION: The decrease of expression of Moesin caused by siRNA plays an important role in human glioma cell growth and invasion, has the potential to become the treatment target molecule of human glioma.
NETL
NSTL
万方数据
维普
