The Effect Of Cyclin-dependent Protein Kinase Subunit 1 On Proliferation, Invasion And Migration Of Glioma Cells

Objective : Cyclin-dependent protein kinase subunit 1 (Cksl), a key regulatory protein in a highly conserved regulation of cell cycle progression. Domestic and international studies have found that Cksl can cause a variety of tumors and lesions,which can be combined with other cyclin-dependent kinases and phosphorylation proteins participate in cell cycle regulation, result in a variety of tumors development,but its less in glioma research. Therefore, this group discussed its expression in glioma and its impact on the biological function of glioma.Methods: First we make the experiment of immunohistochemistry,Immunofluorescence and QRT-PCR, to test the expression of Cksl in normal brain tissue, glioma tissues and normal astrocytes (HA), glioma cells (U87MG) and the location. Then we choose U87MG glioma cell lines for the study, to observe its expression in glioma cells after infected with lentivirus and the effect on glioma.U87MG glioma cell lines were conducted by CkslsiRNA lentivirus, and made the experiment of QRT-PCR, Western Blot to verify its successful infection. To make sure its empact on glioma proliferation, we choose the experiment of cck8 and colony formation assay. The Transwell technology and cell scratch experiment were made to explore the change of glioma invasion and migration, Western Blot test to observe the change of MEK, ERK and phosphorylation-ERK, et al.Results: First we make the experiment of immunohistochemistry and QRT-PCR,the result shoe that Cksl was expressed and localized in the nucleus. The level of Cksl in glioma cells was low than in normal glial cells.The result showed that Cksl were expressed both in normal brain tissue and glioma tissues, and its expression in glioma cell lines was decreased when it compared to normal glial cells. After lentivirus infection, QRT-PCR, Western Blot test to detect the infection efficiency, the results showed that the group of siCks1 1 #,siCks1 3 # have high suppressing efficiency when it compared with siCks1 2#, statistical analysis, the difference was statistically significant (P < 0.05);CCK-8, colony formation (colony)results showed that the downregulation of cksl enhanceed tumor cell proliferation; at the same time,the transwell techniques and healing assay showed that the downregulation of cks1 enhanceed tumor invasion and migration, the difference was statistically significant (P < 0.05). MEK, ERK expression do not decrease but increase the amount of phosphorylation-ERK expression, the difference was statistically significant (P < 0.05) This may provide new ideas and theoretical basis for us to the cure of glioma.Conclusion:1.Cksl was expressed in glioma cells and localized in the nucleus,at the same time the expression of Cksl in glioma cells was lower than in normal astrocytes.2. Inhibit the expression of Cks1, the proliferation, invasion and migration of glioma cells were enhanced.3. Inhibit the expression of Cksl,the levels of ERK phosphorylation was upregulated,Cksl may promote the proliferation of glioma cells and thus affect glioma by varying the p-ERK.