摘要
目的:探讨成纤维细胞生长因子受体4(FGFR4)基因沉默介导RAS/RAF/MAPK信号通路对口腔鳞癌细胞增殖和凋亡的影响。方法:收集本院2018年4月至2019年8月收治的口腔鳞癌患者32对肿瘤组织样本和癌旁非癌组织,利用免疫组织化学和免疫印迹检测FGFR4蛋白的表达;免疫印迹检测正常口腔上皮细胞HOEC和口腔鳞癌细胞(HSC-2、CAL-27和SACC-83)中FGFR4蛋白的表达水平。通过基因沉默技术下调FGFR4表达,利用qRT-RCR和免疫印迹检测SACC-83细胞的FGFR4表达的变化,克隆形成实验检测SACC-83细胞的增殖能力,流式细胞术检测SACC-83细胞的凋亡和细胞周期,免疫印迹检测SACC-83细胞的RAS/RAF/MAPK信号通路。结果:与癌旁非癌组织相比,口腔鳞癌组织的FGFR4表达升高;与正常口腔上皮细胞HOEC相比,口腔鳞癌细胞(HSC-2、CAL-27和SACC-83)的FGFR4表达量高;下调FGFR4表达可抑制SACC-83细胞的增殖,抑制抗凋亡蛋白Bcl-2的表达,上调凋亡蛋白Bax的表达,促进SACC-83细胞的凋亡,同时抑制周期蛋白P21的表达,使SACC-83细胞停滞于G0/G1期;下调FGFR4表达可抑制RAS、RAF及p-MEK和p-ERK1蛋白的表达。结论:下调FGFR4表达可抑制口腔鳞癌细胞增殖,促进细胞凋亡,使其停滞于G0/G1期,可能与抑制RAS/RAF/MAPK信号通路有关。
Objective:To investigate the effects of FGFR4 gene silencing-mediated RAS/RAF/MAPK signaling pathway on proliferation and apoptosis of oral squamous carcinoma cells.Methods:Thirty-two pairs of tumor tissue samples and paracancerous noncancerous tissues were collected from oral squamous cell carcinoma patients admitted to the Affiliated Hospital of Henan Medical College from April 2018 to August 2019.Oral squamous cell carcinoma tissues and adjacent non-cancerous tissues were obtained to detect the expression of FGFR4 by using immunohistochemistry and Western blotting.Western blotting was used to detect the expression of FGFR4 in normal oral epithelial cells HOEC and oral squamous carcinoma cells(HSC-2,CAL-27 and SACC-83).Down-regulation of FGFR4 was performed by gene silencing technology,and then qRT-PCR and Western blotting were used to detect FGFR4 expression in SACC-83 cells,colony formation assay was used to measure proliferation of SACC-83 cells,flow cytometry was utilized to evaluate apoptosis and cell cycle of SACC-83 cells,Western blotting was used to detect the expression of RAS,RAF,p-MEK and p-ERK1 in SACC-83 cells.Results:Compared with adjacent non-cancerous tissues,FGFR4 expression was increased in oral squamous carcinoma tissues;Compared with the normal oral epithelial cell HOEC,FGFR4 was over-expressed in oral squamous carcinoma cells(HSC-2,CAL-27 and SACC-83);Down-regulation of FGFR4 inhibited proliferation of SACC-83 cells and the expression of anti-apoptotic protein Bcl-2,promoted the expression of apoptotic protein Bax and apoptosis of SACC-83 cells.Moreover,silencing FGFR4 inhibited the expression of cyclin P21,and resulted in a higher proportion of SACC-83 cells in G0/G1 phase.Down-regulation of FGFR4 could inhibit the protein level of RAS,RAF,p-MEK and p-ERK1.Conclusion:Down-regulation of FGFR4 can inhibit proliferation of oral squamous carcinoma cells,and promote apoptosis and make them stagnate in G0/G1 phase,which may be related to the inhibition of RAS/RAF/MAPK signaling pathway.
作者
董庆旭
赵郑莉
Dong Qingxu;Zhao Zhengli(Department of Stomatology,Affiliated Hospital of Henan Medical College,Zhengzhou 451100,China)
出处
《解剖学杂志》
CAS
2021年第4期312-318,共7页
Chinese Journal of Anatomy
基金
河南省医学科学研究重点课题计划(202102310399)。
