Siglec-15对胃癌细胞系MKN-45增殖和凋亡作用的影响  

The Effect of Siglec-15 on the Proliferation and Apoptosis of Gastric Cancer Cell Line MKN-45

作　　者：丁祥林 张光波[2] 谢金晶 DING Xiang-lin;ZHANG Guang-bo;XIE Jin-jing(Suzhou Yongding Hospital,Suzhou,Jiangsu,215200,China;Clinical Immunology Institute,First Affiliated Hospital of Soochow University,Suzhou,Jiangsu,215006,China)

机构地区：[1]苏州永鼎医院,江苏苏州215200 [2]苏州大学附属第一医院临床免疫所,江苏苏州215006

出　　处：《中国血液流变学杂志》2022年第3期333-338,F0002,共7页Chinese Journal of Hemorheology

基　　金：苏州市吴江区“科教兴卫”项目(WWK202011)。

摘　　要：目的探讨Siglec-15对胃癌细胞增殖和凋亡的影响。方法免疫组织化学检测胃癌组织中Siglec-15的表达;收集胃癌细胞系检测细胞中Siglec-15的表达,慢病毒感染法构建Siglec-15低表达稳转细胞系(LV-sh-S15)和阴性对照组(LVNC),RT-qPCR和Western blotting实验验证慢病毒转染效率;CCK-8法检测MKN-45细胞系LV-NC组和LV-sh-S15组细胞增殖差异,细胞克隆形成检测Siglec-15对细胞克隆形成能力的影响;流式细胞术检测两组细胞周期及凋亡变化;Western blotting检测凋亡相关蛋白及信号通路蛋白变化。结果免疫组织化学检测发现,相比胃息肉和胃癌早期标本,Siglec-15在胃癌中期标本中表达升高;检测胃癌细胞系中Siglec-15的表达发现,MKN-45细胞中Siglec-15的表达处于中等水平,慢病毒感染法成功构建Siglec-15低表达稳转细胞系;CCK-8和细胞克隆形成实验发现,相比LV-NC组细胞,LV-sh-S15组细胞增殖能力和细胞克隆率降低。流式细胞术和Western blotting发现,相比LV-NC组细胞,LV-sh-S15组细胞凋亡率升高,并且PI3K和AKT的磷酸化水平降低。结论在胃癌早期标本中Siglec-15的表达升高,敲低Siglec-15表达影响MKN-45细胞的增殖和凋亡水平,表明Siglec-15可以作为一种肿瘤标志分子,调控肿瘤进展。Objective To explore the effect of Siglec-15 on the proliferation and apoptosis of gastric cancer cells.Methods Immunohistochemistry was used to detect the expression of Siglec-15 in gastric cancer tissues.Gastric cancer cell lines were collected to detect the expression of Siglec-15 in the cells,and a low-expression stable Siglec-15 cell line(LV-sh-S15)and negative control group(LV-NC)were constructed by lentiviral infection,RT-qPCR and Western blotting experiments were used to verify the efficiency of lentivirus transfection.CCK-8 method was used to detect the difference in cell proliferation of MKN-45 cell line LV-NC group and LV-sh-S15 group,cell clone formation was used to test the effect of Siglec-15 on cell clone formation ability.Flow cytometry was used to detect cell cycle and apoptosis changes in the two groups.Western blotting was used to detect changes in apoptosis-related proteins and related signal pathways.Results Immunohistochemical detection found that compared with gastric polyps and early gastric cancer specimens,Siglec-15 expression was increased in mid-stage gastric cancer specimens.Detection of Siglec-15 expression in gastric cancer cell lines found that the expression of Siglec-15 in MKN-45 cells was at a higher level.At a moderate level,the lentiviral infection method successfully constructed a low-expression and stable Siglec-15 cell line.CCK-8 and cell clone formation experiments found that compared with LV-NC group cells,LV-sh-S15 group cell proliferation ability and cell cloning rate decreased.Flow cytometry and Western blotting showed that compared with the LV-NC group,the apoptosis rate of the LV-sh-S15 group increased,and the phosphorylation level of PI3K and AKT decreased.Conclusion The expression of Siglec-15 is elevated in early gastric cancer specimens.Knockdown of Siglec-15 expression affects the proliferation and apoptosis of MKN-45 cells,indicating that Siglec-15 can be used as a tumor marker molecule to regulate tumor progression.

关 键 词：Siglec-15 胃癌 MKN-45 AKT 

分 类 号：R730.3[医药卫生—肿瘤]