抗癌生物活性肽对与肿瘤相关成纤维细胞共培养的食管癌细胞的增殖影响  

Effect of Anticancer Bioactive Peptides on the Proliferation of Esophageal Cancer Cells Co-Cultured with Cancer-Associated Fibroblasts

作　　者：梁亚冰 杨凌[1] 张满[2] 苏秀兰[1] LIANG Yabing;YANG Ling;ZHANG Man;SU Xiulan(Inner Mongolia Key Laboratory of Medical Cell Biology,Clinical Medical Research Center,the Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010050,China;Department of Thoracic Surgery,the Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010050,China)

机构地区：[1]内蒙古医科大学附属医院临床医学研究中心,内蒙古医学细胞生物学重点实验室,呼和浩特010050 [2]内蒙古医科大学附属医院胸外科,呼和浩特010050

出　　处：《中国细胞生物学学报》2021年第10期1944-1951,共8页Chinese Journal of Cell Biology

基　　金：内蒙古自治区科技成果转化项目(批准号:cgzh2018149);国家自然科学基金(批准号:81341125);内蒙古自治区自然科学基金(批准号:2021lhms08041)资助的课题。

摘　　要：该研究旨在探讨肿瘤相关成纤维细胞(CAFs)对食管鳞状细胞癌(ESCC)细胞增殖的影响及抗癌生物活性肽(ACBP)对增殖的干预作用和可能的作用机制。收集5例手术切除患者的食管癌组织,分离培养得到ESCC CAFs并进行特征指标鉴定;建立CAFs与ESCC细胞共培养体系,CFSE染色和流式细胞术检测细胞增殖;收集CAF-1的条件培养基,加入ACBP进行联合培养KYSE140细胞,IncuCyte检测细胞增殖;qRT-PCR和Western blot检测Hedgehog信号通路相关基因的表达水平。该研究成功分离CAFs,Western blot结果显示CAFs均表达波形蛋白(Vimentin)及纤维连接蛋白(Fibronectin),不表达E-钙黏蛋白(E-cadherin);相比于单独培养,与CAF-1共培养的KYSE140细胞的CFSE水平降低,细胞融合率增加(P<0.05),细胞中的GLI1和PTCH1的mRNA和蛋白水平升高(P<0.05);相比于条件培养基组,ACBP加入可以使KYSE140细胞的融合率下降(P<0.05);相较于单独培养,ACBP加入后KYSE140细胞的GLI1和PTCH1的mRNA和蛋白水平显著下降(P<0.05)。该研究表明,CAFs可以通过激活Hedgehog信号通路促进食管癌细胞增殖,ACBP可在与CAFs共培养条件下通过抑制Hedgehog信号通路抑制食管癌细胞的生长。This work was to investigate the effect of CAFs(cancer-associated fibroblasts)on the proliferation of ESCC(esophageal squamous cell carcinoma)cells and the intervention effect of proliferation and possible mechanism of ACBP(anti-cancer biological active peptide).Five cases of esophageal cancer tissues were collected,from which ESCC CAFs were isolated and cultured,and then the characteristic indexes were identified;the co-culture system of CAFs and ESCC cells was established.CFSE staining and flow cytometry were used to detect cell proliferation;the conditioned medium of CAF-1 was collected,and ACBP was added to co-cultured KYSE140 cells;cell proliferation was detected by IncuCyte,and the expression levels of Hedgehog signaling pathway-related genes were detected by qRT-PCR and Western blot.CAFs were successful separated.The results of Western blot showed that CAFs expressed Vimentin and Fibronectin without E-cadherin.Compared with the control group,the CFSE level of KYSE140 cells decreased and the fusion rate increased(P<0.05),and the mRNA and protein levels of GLI1 and PTCH1 were increased(P<0.05)after co-cultured with CAF-1;compared with the conditional medium group,the fusion rate of KYSE140 cells decreased(P<0.05)after ACBP was added;compared with the control group,the mRNA and protein levels of GLI1 and PTCH1 of KYSE140 cells decreased after ACBP was added(P<0.05).This study suggests that CAFs can promote the proliferation of esophageal cancer cells by activating the Hedgehog signaling pathway,and ACBP can inhibit the growth of esophageal cancer cells by inhibiting the Hedgehog signaling pathway under the co-culture with CAFs.

关 键 词：肿瘤相关成纤维细胞 食管鳞状细胞癌细胞 共培养 抗癌生物活性肽 HEDGEHOG 增殖 

分 类 号：R735.1[医药卫生—肿瘤]