Siglec-15调节STAT1/STAT3通路促进肝癌细胞增殖和迁移的研究  

Siglec-15 regulates the STAT1/STAT3 pathway to promote the proliferation and migration of hepatocellular carcinoma cells

作　　者：闫家伟 关璐璐 郭彩茹 王金艳 张亚丽 文学军 金自立 亓民[2,4] YAN Jia-wei;GUAN Lu-lu;GUO Cai-ru;WANG Jin-yan;ZHANG Ya-li;WEN Xue-jun;JIN Zi-li;QI Min(Xinxiang Medical University GraduateSchool,Xinxiang 453003,Henan Province,China;Key Laboratory of Individualized Liver Cancer Treatment,Luoyang 471000,Henan Province,China;Henan International Joint Laboratory of Tumor Cellular Immunity and Regenerative Medicine,Luoyang 471000,Henan Province,China;Department of Geriatrics,Luoyang Central Hospital,Luoyang 471000,Henan Province,China;Key Laboratoryof Regenerative Medicine,Universityof Virginia,Charlottesville 22093,USA)

机构地区：[1]新乡医学院研究生院,河南新乡453003 [2]洛阳市中心医院洛阳市肝癌个体化治疗重点实验室,河南洛阳471000 [3]洛阳市中心医院河南省肿瘤细胞免疫与再生医学国际联合实验室,河南洛阳471000 [4]洛阳市中心医院老年医学科,河南洛阳471000 [5]美国弗吉尼亚大学再生医学重点实验室,美国夏洛茨维尔市22093

出　　处：《中国临床药理学杂志》2025年第3期350-354,共5页The Chinese Journal of Clinical Pharmacology

基　　金：河南省医学科技攻关计划联合共建基金资助项目(LHGJ20230818)。

摘　　要：目的探究唾液酸结合性免疫球蛋白样凝集素-15(Siglec-15)是否通过调节信号传导与转录激活因子1/3(STAT1/STAT3)通路促进肝细胞癌的增殖和迁移。方法将SMMC-7721细胞分为S-NC组(NC siRNA转染至SMMC-7721细胞)和S-si-Siglec-15-3组(Siglec-15-3 siRNA转染至SMMC-7721细胞),将BEL-7402细胞分为B-NC组(NC siRNA转染至BEL-7402细胞)和B-si-Siglec-15-3组(Siglec-15-3 siRNA转染至BEL-7402细胞)。用噻唑蓝(MTT)法检测细胞的增殖能力;用Transwell小室和划痕试验检测细胞的纵向和横向迁移能力;用蛋白质印迹法检测细胞中STAT3、磷酸化STAT3(p-STAT3)、STAT1、磷酸化STAT1(p-STAT1)、Siglec-15蛋白相对表达水平。结果S-NC组、S-si-Siglec-15-3组、B-NC组、B-si-Siglec-15-3组在24 h时的细胞活力值分别为0.93±0.01、0.87±0.03、0.96±0.03、0.83±0.02;细胞纵向迁移率分别为100.00%、36.40%、100.00%、52.20%;24 h的细胞划痕愈合率分别为(0.76±0.02)%、(0.53±0.02)%、(0.44±0.02)%、(0.36±0.03)%;STAT3蛋白相对表达水平分别为1.11±0.05、0.66±0.05、1.07±0.14、0.49±0.04;p-STAT3蛋白相对表达水平分别为1.07±0.04、0.16±0.06、0.78±0.05、0.32±0.03;STAT1蛋白相对表达水平分别为1.32±0.04、0.71±0.04、0.91±0.03、0.61±0.04;p-STAT1蛋白的表达水平分别为0.99±0.04、0.29±0.05、0.67±0.04、0.20±0.06;Siglec-15蛋白相对表达水平分别为1.51±0.04、0.93±0.05、0.73±0.02、0.12±0.01。S-si-Siglec-15-3组与S-NC组比较,在统计学上差异均有统计学意义(P<0.05,P<0.01,P<0.001);B-si-Siglec-15-3组与B-NC组比较,在统计学上差异均有统计学意义(P<0.05,P<0.01,P<0.001)。结论Siglec-15通过STAT1/STAT3通路促进肝癌细胞的增殖、迁移能力。Objective Probing whether salivary acid-binding immunoglobulin-like agglutinin-15(Siglec-15)promotes hepatocellular carcinoma proliferation and migration by regulating the signal transducer and activator of transcription 1/3(STAT1/STAT3)pathway.Methods SMMC-7721 and BEL-7402 cells were divided into empty vector groups:S-NC and B-NC groups(NC siRNA was transfected into SMMC-7721 and BEL-7402 cell,respectively),Siglec-15 knockdown expression group:S-si-Siglec-15-3 and B-si-Siglec-15-3 groups(Siglec-15-3 siRNA was transfected into SMMC-7721 and BEL-7402 cell,respectively).The proliferation ability of the cells were detected by methyl thiazolyl tetrazolium(MTT);the longitudinal and lateral migration ability of the cells were detected by Transwell and scratch assay;the expression of STAT3,phosphorylation of STAT3(p-STAT3),STAT1,phosphorylation of STAT1(p-STAT1),Siglec-15 was detected by Western blot.Results Cell viability values at 24 h in the S-NC group,S-si-Siglec-15-3 group,B-NC group and B-si-Siglec-15-3 group were 0.93±0.01,0.87±0.03,0.96±0.03 and 0.83±0.02,respectively;the vertical migration rates of cells were 100.00%,36.40%,100.00%and 52.20%,respectively;the healing rate of cell scratches at 24 h were(0.76±0.02)%,(0.53±0.02)%,(0.44±0.02)%and(0.36±0.03)%,respectively;STAT3 protein expression levels were 1.11±0.05,0.66±0.05,1.07±0.14 and 0.49±0.04,respectively;p-STAT3 protein expression levels were 1.07±0.04,0.16±0.06,0.78±0.05 and 0.32±0.03,respectively;STAT1 protein expression levels were 1,32±0.04,0.71±0.04,0.91±0.03 and 0.61±0.04,respectively;p-STAT1 protein expression levels were 0.99±0.04,0.29±0.05,0.67±0.04 and 0.20±0.06,respectively;Siglec-15 protein expression levels were 1.51±0.04,0.93±0.05,0.73±0.02 and 0.12±0.01,respectively.The differences between the S-si-Siglec-15-3 group and the S-NC group,between the B-si-Siglec-15-3 group and the B-NC group were statistically significant in all comparisons(P<0.05,P<0.01,P<0.001).Conclusion Siglec-15 promotes the proliferation and mig

关 键 词：唾液酸结合性免疫球蛋白样凝集素-15 信号传导与转录激活因子1/3通路 肝癌 增殖 迁移 

分 类 号：R97[医药卫生—药品]