FASTKD4在骨肉瘤组织中表达及对骨肉瘤细胞增殖和凋亡的影响

The expression of FASTKD4 in osteosarcoma and its effect on proliferation and apoptosis of osteosarcoma cells

目的探讨FASTKD4在骨肉瘤组织中表达及其对骨肉瘤细胞增殖和凋亡的影响。方法通过免疫组化和qPCR的方法检测FASTKD4在骨肉瘤及癌旁组织中的表达情况,同时还通过qPCR检测FASTKD4在不同骨肉瘤细胞中的表达情况,根据结果选择Saos-2细胞作为研究对象,细胞转染FASTKD4小干扰RNA(shFASTKD4组)及对照阴性序列(shCtrl组)。qPCR和Western blot检测转染后细胞中FASTKD4 mRNA和蛋白表达水平。高内涵筛选和CCK-8检测细胞增殖,基质胶成球实验检测细胞克隆形成能力,流式细胞仪检测细胞凋亡。结果免疫组化和qPCR结果显示FASTKD4在骨肉瘤组织中表达水平高于癌旁组织(P<0.05),与shCtrl组比较,shFASTKD4组骨肉瘤增殖能力受到抑制(P<0.05),且shFASTKD4组骨肿瘤细胞形成的克隆球直径小于shCtrl组(P<0.05)。shFASTKD4组骨肿瘤细胞凋亡率高于shCtrl组(P<0.05)。结论FASTKD4在骨肉瘤组织中高表达,沉默FASTKD4可以抑制骨肉瘤细胞增殖和克隆能力,并促进肿瘤细胞凋亡,进一步研究显示FASTKD4敲除使PI3K/AKT信号通路失活。

Objective To investigate the effect of the expression of FASTKD4 in osteosarcoma and its effect on proliferation and apoptosis of osteosarcoma cells.Methods The levels of FASTKD4 mRNA were detected by qPCR,and protein was detected by immunohistochemistry in osteosarcoma and paracancerous tissues.The levels of FASTKD4 RNA were detected by qPCR in different osteosarcoma cells,and the osteosarcoma cell Saos-2 was used as the research object.Saos-2 cells were transfected with FASTKD4 small interfering RNA(shFASTKD4 group)and negative control(shCtrl group).qPCR and Western blot were used to detect the FASTKD4 mRNA and protein in transfected Saos-2 cells.The proliferation of transfected Saos-2 cells were detected by multiparametric high-content screening and CCK-8.The colony formation ability and apoptosis of transfected Saos-2 cells were detected by clone formation assay and flow cytometry.Reuslts The results of qPCR and immunohistochemistry showed the expression of the FASTKD4 in osteosarcoma tissues was higher than that in paracancerous tissues(P<0.05).The proliferation and colony formation ability of shFASTKD4 group were suppressed when compared with shCtrl group(P<0.05).The percentage of apoptosis in shFASTKD4 group was higher than that in shCtrl group(P<0.05).Conculsion This study confirmed that FASTKD4 was highly expressed in osteosarcoma tissues,and silencing of FASTKD4 could inhibit proliferation,colony formation of osteosarcoma cell and induce the apoptosis of osteosarcoma cell,We also found that FASTKD4 Knockdown inactivated PI3K/AKT Signaling pathway in osteosarcoma cells.