肿瘤间质成纤维细胞对体外培养唾液腺多形性腺瘤细胞生物学行为的影响  

Effect of tumor-stromal fibroblasts on the biological behavior of salivary gland pleomorphic adenoma cells in vitro

作　　者：侯亚丽[1] 李荷香[1] 宋鹏[1] 杨艳霄 郝亚丽 刘慧娟 Hou Yali;Li Hexiang;Song Peng;Yang Yanxiao;Hao Yali;Liu Huijuan(Dept.of Pathology,Hebei Key Laboratory of Stomatology,Hebei Clinical Research Center for Oral Diseases,School and Hospital of Stomatology,Hebei Medical University,Shijiazhuang 050017,China;Key Laboratory of Stomatology,Hebei Key Laboratory of Stomatology,Hebei Clinical Research Center for Oral Diseases,School and Hospital of Stomatology,Hebei Medical University,Shijiazhuang 050017,China)

机构地区：[1]河北医科大学口腔医(学)院病理科,河北省口腔医学重点实验室,河北省口腔疾病临床医学研究中心,石家庄050017 [2]河北医科大学口腔医(学)院重点实验室,河北省口腔医学重点实验室,河北省口腔疾病临床医学研究中心,石家庄050017

出　　处：《华西口腔医学杂志》2023年第2期149-156,共8页West China Journal of Stomatology

基　　金：河北省医学科学研究课题(20210349);河北医科大学大学生创新性实验计划项目(USIP2021110)。

摘　　要：目的探讨肿瘤间质成纤维细胞(TSF)对体外培养唾液腺多形性腺瘤(SPA)细胞增殖、侵袭和迁移的影响。方法采用组织块培养法培养唾液腺多形性腺瘤细胞(SPAC)、TSF和瘤旁正常成纤维细胞(NF),并通过免疫细胞化学染色法进行细胞鉴定。选取对数期TSF、NF制备条件培养液,培养SPAC并命名为TSF-SPAC组和NF-SPAC组,单纯培养SPAC作为对照组(SPAC组)。采用MTT实验、Transwell实验和划痕实验分别检测3组细胞的增殖、侵袭和迁移能力;酶联免疫吸附测定(ELISA)检测3组培养液中血管内皮生长因子(VEGF)的表达量。结果免疫细胞化学染色显示:波形蛋白(Vimentin)在TSF和NF中为阳性,而α-平滑肌肌动蛋白(α-SMA)和成纤维细胞活化蛋白(FAP)在NF中为阴性,在TSF中为弱阳性。MTT结果显示:TSF-SPAC组细胞增殖与NF-SPAC组和SPAC组的差异有统计学意义(P<0.05),NF-SPAC组与SPAC组的细胞增殖的差异无统计学意义(P>0.05)。Transwell侵袭实验和迁移实验显示:3组细胞的侵袭和迁移的差异无统计学意义(P>0.05)。ELISA检测结果显示:3组VEGF的表达量的差异均无统计学意义(P>0.05)。结论TSF可能参与了SPA的生物学行为,促进体外培养的SPAC增殖,对其体外侵袭和迁移无促进作用,为SPA寻找新的治疗靶点提供了方向。Objective This study aims to investigate the effects of tumor-stromal fibroblasts(TSFs)on the proliferation,invasion,and migration of salivary gland pleomorphic adenoma(SPA)cells in vitro.Methods Salivary gland pleomorphic adenoma cells(SPACs),TSFs,and peri-tumorous normal fibroblasts(NFs)were obtained by tissue primary culture and identified by immunocytochemical staining.The conditioned medium was obtained from TSF and NF in logarithmic phase.SPACs were cultured by conditioned medium and treated by TSF(group TSF-SPAC)and NF(group NF-SPAC).SPACs were used as the control group.The proliferation,invasion,and migration of the three groups of cells were detected by MTT,transwell,and scratch assays,respectively.The expression of vascular endothelial growth factor(VEGF)in the three groups was tested by enzyme linked immunosorbent assay(ELISA).Results Immunocytochemical staining showed positive vimentin expression in NF and TSF.Results also indicated the weak positive expression ofα-smooth muscle actin(SMA)and fibroblast activation protein(FAP)in TSFs and the negative expression ofα-SMA and FAP in NFs.MTT assay showed that cell proliferation in the TSF-SPAC group was significantly different from that in the NF-SPAC and SPAC groups(P<0.05).Cell proliferation was not different between the NF-SPAC and SPAC groups(P>0.05).Transwell and scratch assays showed no difference in cell invasion and migration among the groups(P>0.05).ELISA showed that no significant difference in VEGF expression among the three groups(P>0.05).Conclusion TSFs may be involved in SPA biological behavior by promoting the proliferation of SPACs but has no effect on the invasion and migration of SPACs in vitro.Hence,TSF may be a new therapeutic target in SPA treatment.

关 键 词：唾液腺多形性腺瘤 肿瘤间质成纤维细胞 增殖 侵袭 迁移 交界性肿瘤 

分 类 号：Q2[生物学—细胞生物学]