cPLA2抑制剂对EGF和FBS诱导的Hela细胞增殖及cPLA2活性和蛋白表达的影响

Cytosolic phospholipase A_2 involved in epidermal growth factor and fetal bovine serum-induced proliferation in Hela cells and its activity and expression

目的:研究抑制胞浆型磷脂酶A2(cPLA2)对表皮生长因子(EGF)和胎牛血清(FBS)诱导Hela细胞增殖及cPLA2活性和蛋白表达的影响。方法:采用cPLA2特异性抑制剂AACOCF3、反义寡核苷酸(SK7111)及其上游酶ERK1/2抑制剂U0126,分别作用于Hela细胞,观察在EGF及FBS作用下Hela细胞增殖以及细胞周期的变化;采用cPLA2活性检测盒及Western blot检测cPLA2活性及表达。结果:AACOCF3、SK7111及U0126均能明显抑制cPLA2活性,且呈剂量依赖性地抑制EGF和FBS诱导的Hela细胞增殖,尤以SK7111作用最为明显;流式细胞仪检测细胞周期表明,AACOCF3和U0126可将Hela细胞阻滞在S期。结论:cPLA2选择性抑制剂及其反义寡核苷酸可通过抑制cPLA2活性显著抑制EGF和FBS诱导Hela细胞增殖,同时,ERK1/2在此过程中可能具有调节作用。

Objective：To explore effects of cytosoloic phospholipase A2（cPLA2） on epidermal growth factor（EGF） induced and fetal bovine serum（FBS） induced Hela cells proliferation and its activity and expression.Methods：We used inhibitor AACOCF3 and antisense oligodeoxynucleotide（ODN） of cPLA2,and inhibitor U0126 of ERK1/2 to inhibit cPLA2 activity and expression,and observed the changes of Hela cells viabilities and cell cycle by CellTiter-Blue and flow cytometry.Meanwhile,cPLA2 activity and expression were also detected by cPLA2 activity assay kit and western blot.Results：Both inhibitor and antisense ODN of cPLA2 suppressed activity of cPLA2,and obviously inhibited the proliferation induced by EGF and FBS in dose-dependent manner in Hela cells.Both inhibitors of cPLA2 and ERK1/2 decreased cell proportion of G2/M obviously,and they blocked Hela at the S/G2 boundary.Moreover,the inhibitory effect on Hela cells proliferation of antisense ODN was most obvious and the inhibitory effect on cPLA2 protein expression was in a dose-dependent manner.The similar situation of ERK1/2 inhibitor was observed.Conclusion：cPLA2 could play important roles in EGF- and FBS-induced Hela cells proliferation,and ERK1/2 probably contribute to regulation on this course.