NCAPG促进成骨肉瘤细胞增殖的研究

Study of the proliferation of osteosarcoma cells promoted by NCAPG

目的:初步探究非染色体结构维护亚基凝聚素Ⅰ复合物亚基G(NCAPG)基因对成骨肉瘤细胞增殖的影响。方法:选择商丘市第一人民医院和烟台毓璜顶医院2010年7月—2021年1月收治的成骨肉瘤患者58例,通过免疫组织化学染色法对成骨肉瘤和正常组织中NCAPG基因的表达水平进行比较。成骨肉瘤细胞MG-63和U-2 OS,通过转染慢病毒载体和对照载体的shRNA,敲减NCAPG基因来抑制其表达,qRT-PCR、Western印迹验证敲减效果,通过集落形成实验、CCK-8检测其对成骨肉瘤细胞MG-63与U-2 OS细胞增殖的影响。结果:免疫组织化学染色显示,与正常组织相比,成骨肉瘤组织中NCAPG蛋白表达水平明显升高,并且NCAPG基因的表达与临床分期(χ^(2)=6.375,P=0.042)和肿瘤大小(χ^(2)=4.169,P=0.012)密切相关。qRT-PCR、Western印迹显示,慢病毒载体明显抑制了NCAPG mRNA及蛋白的表达(mRNA:tUG-63=20.7,tU-2 OS=22.9;蛋白质:tUG-63=18.7,tU-2 OS=16.9,均P<0.001)。与NCAPG基因未敲减相比,NCAPG基因敲减后MG-63与U-2 OS克隆形成细胞数量降低(tUG-63=18.3,tU-2 OS=15.9,均P<0.01)。结论:成骨肉瘤组织NCAPG基因表达增加,敲减NCAPG基因可抑制成骨肉瘤细胞的增殖。

Objective:To preliminarily explore the effect of non-structural maintenance of chromosomes condensation I compex subunit G(NCAPG)gene on the proliferation of osteosarcoma cell.Methods:A total of 58 patients with osteosarcoma admitted to Shangqiu First People′s Hospital and Yantai Yuhuangding Hospital from July 2010 to January 2021 were selected to compare the expression level of NCAPG gene in osteosarcoma and normal tissues by immunohistochemical staining.The expression of NCAPG gene in osteosarcoma cells mG-63 and U-2 OS was inhibited by transfection of shRNA of lentiviral vector and control vector,and the knockdown effect was verified by qRT-PCR and Western blotting.Colony formation assay and CCK-8 assay were used to detect the effects on proliferation of oste osarcoma cells MG-63 and U-2 OS.Results:Immunohistochemical staining showed that the expression of NCAPG protein in osteosarcoma tissues was significantly higher than that in normal tissues,and the expression of NCAPG gene was closely correlated with clinical stage(Х^(2)=6.375,P=0.042)and tumor size(Х^(2)=4.169,P=0.012).qRT-PCR and Western blotting showed that the expression of NCAPG mRNA and protein was significantly inhibited by lentiviral vector(mRNA:tUG-63=20.7,tU-2 OS=22.9;protein:tUG-63=18.7,tU-2 OS=16.9,P<0.001).Compared with no NCAPG gene knockdown,the number of mG-63 and U-2 OS clones was decreased after NCAPG gene knockdown(tUG-63=18.3,tU-2 OS=15.9,P<0.01).Conclusion:The expression of NCAPG gene in osteosarcoma tissue is increased,and the knocking down NCAPG gene can inhibit the proliferation of osteosarcoma cells.