口腔鳞癌对牙龈成纤维细胞转化为肿瘤相关成纤维细胞的影响  

The influence of oral squamous cell carcinomas on normal fibroblasts turning into oral carcinoma associated fibroblasts

作　　者：孟琳 王丹丹[1,2] 布文奂 李杏[1,2] 辛颖 王璐[1,2] 孙宏晨 

机构地区：[1]吉林省牙发育及颌骨重塑与再生重点实验室,吉林长春130021 [2]吉林大学口腔医院病理科,吉林长春130021

出　　处：《口腔生物医学》2017年第3期136-140,共5页Oral Biomedicine

基　　金：国家自然科学基金资助项目(81320108011);吉林省省级产业创新专项资金项目(2016C044-3);吉林省科技发展计划项目(20170101093JC)

摘　　要：目的:探讨肿瘤相关成纤维细胞(CAFs)的来源。方法:分离正常牙龈组织和口腔鳞癌组织的成纤维细胞,将口腔鳞状细胞癌细胞Cal27培养基加入到正常牙龈成纤维细胞内(NFs);以NFs作为阴性对照、来自于口腔鳞癌组织的成纤维细胞为阳性对照,通过免疫细胞化学染色、实时定量RT-PCR等技术对其进行对比研究。结果:与NFs相比,CAFs胞体更大,排列更加混乱,生长更为快速,形态明显不同。免疫细胞化学染色结果显示:CAFs波形蛋白(VIM)呈阳性,α-平滑肌肌动蛋白(α-SMA)部分呈强阳性、部分呈阴性,成纤维细胞激活蛋白(FAP)呈强阳性,血小板源性生长因子受体α(PDGFR-α)呈阳性,细胞角蛋白(CK)呈阴性;NFs的VIM呈阳性,其余呈阴性;NFs与Cal27条件培养基共培养后,可见α-SMA表达部分阳性,FAP和PDGFR-α表达呈强阳性。实时定量RT-PCR结果发现:与NFs比较,CAFs、NFs与Cal27条件培养基共培养后α-SMA、FAP的表达明显上调,PDGFR-α的表达明显下调。结论:口腔鳞状细胞癌细胞可激活NFs转化为CAFs,NFs可能为CAFs的来源之一。Objective： To investigate the source of cancer associated fibroblasts（ CAFs）. Methods： Normal fibroblasts（ NFs） and CAFs were separated from normal gingival tissues and oral squamous carcinoma tissues,and then Cal27 medium was addedto NFs; we carried on the contrast research by immunohistochemistry,qRT-PCR technology,regarding NFs as negative control,CAFs as the positive control. Results： Compared with NFs,CAFs cell body was bigger,more disarranged,more rapid in growth,and with obviously different morphology. Cell immunohistochemistry showed： CAFs showed positive for vimentin,α-SMA was partly strongly positive,partly negative and FAP was strong positive,PDGFR-α was positive,CK（ total） was negative. NFs was positive for vimentin,but the rest were negative. After culturing NFs with conditioned medium of Cal27,we found α-SMA was partly strongly positive,FAP was strong positive and PDGFR-α was positive. qRT-PCR showed： compared with NFs,α-SMA,FAP mRNA expression in CAFs and CMNFs was significantly higher,and PDGFR-α mRNA expression was significantly lowered. Conclusions： Oral squamous cell cancer cells can activate the NFs to CAFs,NFs likely to be a source of CAFs.

关 键 词：肿瘤相关成纤维细胞 口腔鳞状细胞癌 激活机制 

分 类 号：R739.8[医药卫生—肿瘤]