ABSTRACT
Objective To construct PES1 shRNA stable expression cell lines in tongue squamous cell carcinoma ( TSCC) cells and to study the effect of knockdown of PES 1 on the growth of TSCC cells .Methods Recombinant lentivirus carrying PES1 shRNA was packaged and obtained in 293T cells.TSCC cells (Tca8113, SCC6 and SCC15) were infected with the lentivirus and selected for stable cells .PES1 expression was identified by Western blot .The effect of inhibition of PES1 on the growth and cell cycle of TSCC cells was detected by growth curve and flow cytometry .Results TSCC cells stably expressing PES1 shRNA were constructed.Knockdown of PES1 inhibited cell proliferation and induced cell cycle ar-rest at G0/G1 phase.Knockdown of PES1 inhibited expression of cyclin D1 in TSCC cells.Conclusion Inhibition of PES1 results in reduced cell proliferation , cell cycle arrest at G 0/G1 phase and reduction of cyclin D 1 expression in TSCC cells . PES1 may be a target for TSCC gene therapy .
ABSTRACT
Objective To investigate the expression of PES1 in ovarian cancer and its relationship with the vascular endothelial growth factor(VEGF) expression .Methods The expression of PES1 in ovarian cancer tissues and corresponding pericancerous tis-sues was detected with Western blot .Results The expression of PES1 in ovarian cancer tissues was obviously higher than that in the pericancerous tissues .The secretion amounts of VEGF in cell culture fluid of CAOV-3 and ES-2 with transfection of FLAG-PES1 were elevated from (178 .0 ± 11 .8) pg/mL and (309 .5 ± 18 .5) pg/mL to (375 .0 ± 18 .3) pg/mL and (633 .2 ± 25 .7)pg/mL respectively (P<0 .01) ,the secretion amounts of VEGF were also increased(P<0 .01) .The relative VEGF mRNA levels were also raised by 1 .8 times and 2 times respectively (P< 0 .01);Western blot simultaneously showed that the over expression of PES1 could increase the expression of HIF-1αin these two kinds of cells .The luciferase report gene transcriptional activation activity de-tection reveald that PES1 had no direct effect on the VEGF transcription .After cotransfecting HIF-1αSiRNA and FLAG-PES1 to CAOV-3 and ES-2 cells ,Western blot demonstrated that HIF-1αSiRNA could obviously inhibit the increase of HIF-1αexpression induced by PES1 ,at the same time the secretion amounts of VEGF and mRNA level were also supressed .Conclusion The expres-sion of PES1 is obviously up-regulated in ovarian cancer tissues .