CDCA5在裸鼠肝细胞癌中的作用及临床价值

The Role and Clinical Value of CDCA5 in Nude Mice Hepatocellular Carcinoma

目的探讨不同程度CDCA5表达在裸鼠肝细胞癌中的作用及临床价值。方法取HepG2正常细胞组、CDCA5低表达组、CDCA5高表达组、HepG2肝癌细胞株,将高表达组、低表达组CDCA5高效转染人肝癌HepG2细胞,并设阴性对照组(NC组)。免疫印迹法检测转染24 h后各组CDCA5蛋白的表达水平,CCK-8法检测转染24、48、72、96、120 h各组细胞的增殖能力,Annexin V-FITC/PI双标记流式细胞术检测低表达组转染48 h的凋亡情况;将HepG2正常细胞组,CDCA5低表达组的细胞分别注射于裸鼠成瘤,观察成瘤时间,第30天处死小鼠,观察瘤体状态。免疫组化检测CDCA5在裸鼠肿瘤中的表达。结果高表达和低表达组的CDCA5蛋白水平均高于NC组(P<0.05);自转染48 h起,高表达组和低表达组的相对增殖率均高于NC组(P<0.05);高表达组和低表达组转染72 h后的相对增殖率分别为(130.03±0.35)%和(70.10±1.04)%,均高于其他观察时间点(P<0.05);低表达组转染48 h的早期和晚期凋亡率均高于NC组(P<0.05);裸鼠皮下注射普通HepG2细胞组及CDCA5低表达HepG2细胞,瘤体重量分别为(0.93±0.06)g、(0.69±0.10)g,差异有统计学意义(P<0.05);CDCA5在低表达组裸鼠肿瘤组织中表达降低。结论CDCA5能促进HepG2癌细胞的增殖,可通过调控CDCA5表达水平,有效抑制肝癌细胞的增殖并促进其凋亡。

Objective To explore the role and clinical value of different levels of CDCA5 expression in nude mice hepatocellular carcinoma.Methods HepG2 normal cell group,CDCA5 low expression group,CDCA5 high expression group and HepG2 hepatocarcinoma cell lines were selected.The high expression group and low expression group CDCA5 were efficiently transfected into human liver cancer HepG2 cells,and a negative control group(NC group)was established.Western blotting was used to detect the expression level of CDCA5 protein in each group after 24 h of transfection.CCK-8 method was used to detect the proliferation ability of cells in each group after transfection 24,48,72,96,120 h.Annexin V-FITC/PI double label flow Cytometry was used to detect the apoptosis of the low expression group for 48 h after transfection;the HepG2 normal cell group and the CDCA5 low expression group were injected into nude mice to form tumors,and the tumor formation time was observed.The mice were killed on the 30th day and the tumors were observed body state.Immunohistochemistry was used to detect the expression of CDCA5 in tumors of nude mice.Results The CDCA5 protein levels of the high and low expression groups were higher than those of the NC group(P<0.05);since 48 h after transfection,the relative proliferation rates of the high and low expression groups were higher than those of the NC group(P<0.05);The relative proliferation rates of high expression group and low expression group 72 h after transfection were(130.03±0.35)%and(70.10±1.04)%,respectively,which were higher than other observation time points(P<0.05);low expression group was transfected the early and late apoptosis rates at 48 h were higher than those in the NC group(P<0.05);the nude mice were subcutaneously injected with ordinary HepG2 cells and CDCA5 low-expressing HepG2 cells,and the tumor weights were(0.93±0.06)g and(0.69±0.10)g,the difference was statistically significant(P<0.05);the expression of CDCA5 in the tumor tissues of nude mice in the low expression group decreased.Conclusion CDCA5 can promote the proliferation of HepG2 cancer cells,and could effectively inhibit the proliferation of liver cancer cells and promote their apoptosis by regulating the expression level of CDCA5.