Siglec-15和PD-L1在肝细胞癌中表达的关系及其对肝癌细胞生物学影响

The relationship between Siglec-15 and PD-L1 expression in hepatocellular carcinoma and its biological impact on liver cancer cells

目的 分析Siglec-15和PD-L1在肝细胞癌(hepatocellutar carcinoma,HCC)中的表达及两者之间的关系,联合临床病理因素、细胞功能实验进行探究。方法 收集2015年6月至2020年6月于郑州大学附属洛阳中心医院行肝癌根治切除术的30例HCC患者癌组织及正常新鲜标本及石蜡切片标本。利用qRT-PCR、Western blotting、组织切片免疫组化检测Siglec-15和PD-L1的表达,并对两者表达的关系进行分析。进一步用慢病毒介导的siRNA及基因转导技术将siRNA-Siglec-15和siRNA-NC(空白质粒)分别转染至SMMC-7721细胞,分为si-Siglec-15组和si-NC组,对照组细胞不做任何处理。利用CCK-8法、划痕实验、Transwell实验检测HCC细胞的增殖、迁移、侵袭。结果 HCC组织中Siglec-15蛋白和PD-L1蛋白的表达均高于癌旁正常组织(P<0.05),且Siglec-15蛋白表达与PD-L1蛋白表达呈负相关(R=-0.586,P<0.001)。构建Siglec-15低表达细胞株,发现si-Siglec-15组HCC细胞SMMC-7721的增殖、迁移、侵袭能力下降。结论 Siglec-15在HCC组织中高表达,与PD-L1的表达呈负相关;Siglec-15低表达可抑制HCC细胞SMMC-7721的增殖、迁移、侵袭。

Objective To analyze the expression of Siglec-15 and PD-L1 in hepatocellutar carcinoma(HCC) tissues and the relationship between them.The clinicopathological factors and cell function test were combined and explored.Methods Fresh liver cancer and paraside carcinoma specimens and paraffin sections were collected from 30 patients with HCC underwent radical resection from Jun.2015 to Jun.2020.The expressions of Siglec-15 and PD-L1 were determined by qRT-PCR,Western blotting,and immunohistochemical experiments of tissue sections.And the relationship between the two expression was analyzed.Further,siRNA-Siglec-15,siRNA-NC(blank plasmid) were transfected into SMMC-7721 cells by lentivirus-mediated siRNA and by-gene transduction.They were divided into si-Siglec-15 group and si-NC group,control group cells were treated without any treatment.Proliferation,migration and invasion experiments were performed using CCK-8,scratch assay and Transwell.Results The expressions of Siglec-15 and PD-L1were higher in HCC patients than those in normal tissues(P<0.05),and Siglec-15 expression was negatively correlated with PD-L1(R=-0.586,P<0.001).Further knocked down Siglec-15 expression in SMMC-7721 cells by using lentiviral infection to construct low Siglec-15 expression cell lines,which found that the proliferation,migration and invasion ability of cells in the test group were decreased.Conclusion Siglec-15 is highly expressed in HCC tissues and significantly negatively associated with PD-L1;low Siglec-15 expression inhibites the proliferation,migration and invasion of SMMC-7721 cells.