PFKFB3激活MAPK信号通路促进卵巢癌增殖和转移

PFKFB3 Activates MAPK Signaling Pathway to Promote the Proliferation and Metastasis of Ovarian Cancer

目的探讨6-磷酸果糖激酶-2/果糖双磷酸酶-2同工酶3(PFKFB3)在卵巢癌组织中的表达及其与卵巢癌临床病理特征之间的关系,并通过体外实验初步探讨PFKFB3的促癌作用及可能的作用机制。方法采用免疫组化法检测45例卵巢癌组织和33例输卵管组织中PFKFB3蛋白的表达,分析PFKFB3蛋白表达与卵巢癌临床病理特征之间的相关性。将卵巢癌SKOV3细胞分为三组:空白组(Blank组)不作任何处理,阴性对照组(si-NC组)转染空载质粒,si-PFKFB3组转染PFKFB3特异性干扰质粒。Western blotting检测PFKFB3蛋白表达验证敲减效率,CCK-8检测各组细胞活力,平板克隆形成实验检测各组细胞克隆形成能力,Transwell实验检测各组细胞的迁移和侵袭能力,Western blotting检测MAPK信号通路蛋白p-p38、t-p38的表达。结果PFKFB3主要在细胞核和细胞浆中表达,且在卵巢癌组织中的阳性表达率显著高于输卵管组织(P<0.05);卵巢癌组织学分级越低,FIGO分期越高,PFKFB3蛋白表达阳性率越高(P<0.05)。与Blank组和si-NC组比较,si-PFKFB3组PFKFB3蛋白表达水平显著降低,细胞活力显著下降,细胞克隆形成能力显著降低,细胞迁移和侵袭能力显著下降,p-p38蛋白表达水平及p-p38/p38比值显著降低(P<0.05)。结论PFKFB3在卵巢癌组织中呈高表达,敲减PFKFB3可抑制卵巢癌的增殖、转移和MAPK信号通路激活。

Objective To investigate the expression of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3(PFKFB3)in ovarian cancer and its relationship with the clinicopathological features,and to explore the carcinogenic effect and possible mechanism of PFKFB3 in vitro.Methods The expression of PFKFB3 in 45 cases of ovarian cancer and 33 cases of fallopian tube tissues were detected by immunohistochemitry technique.The relationship between expression of PFKFB3 and clinicopathological characteristics of ovarian cancer was analyzed.SKOV3 cells were divided into three groups:blank group without any treatment,negative control group(si-NC)transfected with empty plasmid,si-PFKFB3 group transfected with PFKFB3 specific interference plasmid.Western blotting was applied to detect the PFKFB3 protein expression and to verify the knock down efficiency.Besides,the activity of cells in each group was detected by CCK-8,and cell clone formation ability by plate clone formation test,and the cell migration and invasion by transwell test.Finally,the expression of p-p38 and t-p38 protein in MAPK signal pathway was detected by Western blotting.Results PFKFB3 was mainly expressed in nucleus and cytoplasm.Its expression positive rate was higher in ovarian cancer than in fallopian tube tissues,and the difference was statistically significant(P<0.05).Lower histological grade as well as higher FIGO stage were related to higher positive rate of PFKFB3 protein expression(P<0.05).Compared with blank group and si-NC group,the expression of PFKFB3 protein and cell viability in si-PFKFB3 group were significantly decreased.In addition,the ability of cell clone formation as well as cell migration and invasion were significantly reduced(P<0.05).The p-p38 protein expression and p-p38/p38 ratio was also decreased(P<0.05).Conclusion PFKFB3 was highly expressed in ovarian cancer tissues.Knockdown of PFKFB3 could inhibit the proliferation,metastasis and activation of MAPK signal pathway of ovarian cancer.