锌指蛋白22基因对肝癌细胞增殖、凋亡及侵袭转移的影响

Effect of zinc finger protein 22 gene on proliferation,apoptosis,invasion and metastasis of hepatocellular carcinoma cells

目的检测锌指蛋白22(ZNF22)基因在肝癌组织中的表达水平并观察ZNF22对肝癌细胞增殖、凋亡、侵袭、转移能力的影响。方法收集2020年1月至2021年6月在暨南大学附属广州红十字会医院行手术切除的32例肝细胞癌患者的肝癌组织及癌旁肝组织标本。分析ZNF22基因在32例肝癌标本以及癌症基因组图谱数据库371例肝癌样本中的表达水平。构建ZNF22基因敲减的SNU-449和JHH-7肝癌细胞系。通过细胞增殖实验、平板克隆形成实验、细胞凋亡实验、划痕愈合实验、Transwell侵袭实验、裸鼠皮下成瘤、尾静脉注射转移和小动物活体成像实验,观察ZNF22对肝癌细胞的影响。结果ZNF22基因在肝癌组织中表达量高于癌旁组织,差异有统计学意义(P<0.001)。ZNF22敲减的SNU-449和JHH-7细胞的生长速度均低于相应的对照组细胞,差异有统计学意义(均P<0.001)。相比阴性对照组,ZNF22敲减的SNU-449细胞形成的克隆数目减少(26±8比59±5),细胞凋亡水平增加(6.60%±0.22%比2.38%±0.30%),细胞的迁移率降低(14.47%±6.42%比68.84%±8.01%),侵袭细胞数也降低(48.00±2.23比179.00±4.81),差异均有统计学意义(均P<0.05)。ZNF22敲减的JHH-7细胞注射于裸鼠皮下后未见明显肿瘤生长,全身荧光表达量低于阴性对照组,差异有统计学意义(P<0.05),裸鼠解剖观察亦未见转移瘤。结论ZNF22在肝癌组织中的表达升高,ZNF22基因敲减抑制了肝癌细胞的生长、增殖、侵袭、转移和成瘤能力,并可诱导肝癌细胞的凋亡。

Objective To investigate the expression of zinc finger protein 22(ZNF22)gene in hepatocellular carcinoma(HCC)and its effect on tumor proliferation,apoptosis,invasion and metastasis of HCC.Methods The expression of ZNF22 in 32 HCC specimens,and 371 HCC samples from the cancer genome atlas database were analyzed.ZNF22 knockdown and negative control SNU-449 and JHH-7 HCC cell lines were constructed.The effects of ZNF22 on HCC cells were observed by cell proliferation assay,plate clone formation assay,apoptosis assay,scratch healing assay,Transwell invasion assay,subcutaneous tumor formation,tail vein injection transfer,and small animal live imaging assay in nude mice.Results The expression of ZNF22 gene is higher in HCC tissues than in paracellular carcinoma tissues,and the difference was statistically significant(P<0.001).The growth rate of SNU-449 and JHH-7 cells in ZNF22 knockdown group was lower than that in control group,and the difference was statistically significant(P<0.001).Compared with negative control group,the clone number formed by SNU-449 cells in ZNF22 knockdown group decreased(26±8 vs.59±5,P<0.01),the level of apoptosis increased(6.60%±0.22%vs.2.38%±0.30%,P<0.001),the migration rate decreased(14.47%±6.42%vs.68.84%±8.01%,P<0.001),and the number of invasive cells decreased(48.00±2.23 vs.179.00±4.81,P<0.001).There was no obvious tumor growth after subcutaneous injection of JHH-7 cells into nude mice in ZNF22 knockdown group,and the systemic fluorescence expression was lower than that of the negative control group,and the difference was statistically significant(P<0.05).No metastases were observed on autopsy in knockdown group nude mice.Conclusion ZNF22 is highly expressed in HCC while knockdowing ZNF22 gene inhibited the growth,proliferation,invasion,metastasis of HCC cells,and induced apoptosis of HCC cells.