摘要
目的:探讨党参炔苷调节鞘氨醇激酶1(SPHK1)/1-磷酸化鞘氨醇(S1P)/1-磷酸鞘氨醇受体3(S1PR3)信号通路对口腔鳞状细胞癌(OSCC)CAL 27细胞恶性进展的影响。方法:将对数生长期CAL 27细胞分为NC组(0μmol/L党参炔苷)、低剂量党参炔苷组(10μmol/L党参炔苷)、中剂量党参炔苷组(20μmol/L党参炔苷)、高剂量党参炔苷组(40μmol/L党参炔苷)、高剂量党参炔苷+SPHK1激活剂PMA组(40μmol/L党参炔苷+100 nmol/L PMA)。MTT法、平板克隆法、流式细胞术和TUNEL染色分别测定CAL 27细胞活力、克隆形成率及凋亡情况;Transwell实验检测CAL 27细胞的迁移和侵袭;Western blot测定SPHK1/S1P/S1PR3信号通路蛋白及迁移蛋白MMP-2、MMP-9表达。将CAL 27细胞皮下注射到小鼠右侧,建立异种移植物模型。小鼠分为对照组和党参炔苷组,每组6只,治疗结束后,测量肿瘤体积和重量,TUNEL染色检测肿瘤组织细胞凋亡,免疫组化染色检测Ki67和MMP-9在组织中的表达,Western blot测定SPHK1、S1P、S1PR3蛋白在组织中的表达。结果:与NC组相比,低、中、高剂量党参炔苷组CAL 27细胞活力、克隆形成率、细胞迁移数及侵袭数、SPHK1、S1P、S1PR3、MMP-2、MMP-9蛋白表达均降低,细胞凋亡率、TUNEL阳性率升高(P<0.05)。与高剂量党参炔苷组对比,高剂量党参炔苷+PMA组CAL 27细胞活力、克隆形成率、细胞迁移及侵袭数、SPHK1、S1P、S1PR3、MMP-2、MMP-9蛋白表达均升高,细胞凋亡率、TUNEL阳性率降低(P<0.05)。与对照组相比,党参炔苷组肿瘤体积、肿瘤重量、Ki67+和MMP-9+细胞比例及SPHK1、S1P、S1PR3蛋白表达降低,凋亡细胞比例升高(P<0.05)。结论:党参炔苷可能通过抑制SPHK1/S1P/S1PR3信号通路活化进而抑制CAL 27细胞恶性进展。
Objective:To investigate the impact of lobetyolin on malignant progression of oral squamous cell carcinoma(OSCC)CAL 27 cells by regulating sphingosine kinase 1(SPHK1)/sphingosine-1-phosphate(S1P)/sphingosine 1 phosphate receptor 3(S1PR3)signaling pathway.Methods:CAL 27 cells in logarithmic growth phase were divided into NC group(0μmol/L lobetyolin),low dose lobetyolin group(10μmol/L lobetyolin),medium dose lobetyolin group(20μmol/L lobetyolin),high dose lobetyolin group(40μmol/L lobetyolin),and high dose lobetyolin+SPHK1 activator PMA group(40μmol/L lobetyolin+100 nmol/L PMA).Viability,clone formation rate and apoptosis of CAL 27 cells were detected by MTT assay,plate cloning assay,flow cytometry and TUNEL staining,respectively.Migration and invasion of CAL 27 cells were detected by Transwell experiment.Expressions of SPHK1/S1P/S1PR3 signaling pathway proteins and migration protein MMP-2,MMP-9 were detected by Western blot.Xenograft model was established by subcutaneous injection of CAL 27 cells into the right side of mice.The mice were divided into control group and lobetyolin group,with 6 mice in each group.After treatment,tumor volume and weight were measured,apoptosis of tumor tissue was detected by TUNEL staining,expressions of Ki67 and MMP-9 in tissue were detected by immunohistochemical staining,expressions of SPHK1,S1P and S1PR3 proteins in tissue were determined by Western blot.Results:Compared with NC group,viability,clone formation rate,cell migration number,invasion number,and protein expressions of SPHK1,S1P,S1PR3,MMP-2 and MMP-9 of CAL 27 cells in low,medium and high doses lobetyolin groups were decreased,while apoptosis rate and TUNEL positive rate were increased(P<0.05).Compared with high dose lobetyolin group,viability,clone formation rate,cell migration number,invasion number,and protein expressions of SPHK1,S1P,S1PR3,MMP-2 and MMP-9 of CAL 27 cells in high dose lobetyolin+PMA group were increased,while apoptosis rate and TUNEL positive rate were decreased(P<0.05).Compared with control group,tumor volume,tumor weight,propor‐tion of Ki67+and MMP-9+cells and protein expressions of SPHK1,S1P and S1PR3 were decreased in lobetyolin groups,while proportion of apoptotic cells was increased(P<0.05).Conclusion:Lobetyolin may inhibit the malignant progression of CAL 27 cells by inhibiting the activation of SPHK1/S1P/S1PR3 signaling pathway.
作者
王鹏
陈慧
代喆颖
江铃
彭红
WANG Peng;CHEN Hui;DAI Zheying;JIANG Ling;PENG Hong(Department of Stomatology,the Third Hospital of Wuhan,Wuhan 430070,China)
出处
《中国免疫学杂志》
北大核心
2026年第2期341-347,355,共8页
Chinese Journal of Immunology
基金
武汉市卫生健康委员会科研项目(WX18C06)。
