YBX3通过MAPK途径促进鼻咽癌增殖和生长的实验研究

YBX3 Promotes the Proliferation and Tumor Growth of Nasopharyngeal Carcinoma by MAPK Pathway

目的探讨YBX3在鼻咽癌细胞中表达及对鼻咽癌增殖、生长的作用和分子机制。方法应用Q-PCR和Western blot,检测YBX3在多种细胞中的表达水平。选取SiRNA和ShRNA干扰高表达YBX3的5-8F细胞后,结合CFSE、CCK8、克隆形成实验检测YBX3对鼻咽癌细胞的增殖影响;在体小鼠荷瘤实验检测YBX3对肿瘤生长的作用;以及细胞增殖相关信号通路MAPK p38及其下游关键标志物PCNA等蛋白的表达。结果相对于正常鼻咽上皮细胞NP460,YBX3在鼻咽癌细胞中表达显著升高,差异具有统计学意义(P<0.01);且在5-8F细胞中表达最高,差异最明显(P<0.01);采用SiRNA和ShRNA干扰5-8F细胞,干扰效率实验结果显示YBX3表达显著降低,差异具有统计学意义(P<0.01)。CFSE,CCK8和克隆形成等增殖实验结果显示干扰YBX3表达显著降低细胞增殖,差异具有统计学意义(P<0.01);小鼠荷瘤实验结果也显示YBX3-ShRNA组小鼠肿瘤生长显著降低,差异具有统计学意义(P<0.01)。此外,干扰YBX3表达显著抑制MAPK p38的活性和下游关键蛋白PCNA的表达。结论干扰YBX3表达可通过MAPK p38/PCNA途径抑制5-8F细胞增殖,降低鼻咽癌肿瘤生长。

Objective To investigate the expression of YBX3 in nasopharyngeal carcinoma cells,the effect on proliferation and tumor growth of nasopharyngeal carcinoma and the molecular mechanism of YBX3.Methods Q-PCR and Western Blot were used to detect the expression of YBX3 in nasopharyngeal carcinoma cells.After SiRNA and ShRNA were used to interfere the expression of YBX3 of 5-8 F cells with highest expressing YBX3,the effects of YBX3 on cell proliferation of 5-8 F cells were detected by CFSE,CCK8 and Clone formation assays.And the effect of YBX3 on tumor growth of nasopharyngeal carcinoma was detected tumor-bearing experiment in vivo.Moreover,the activity of key proliferation related signaling pathway MAPK p38 and the expression of downstream key protein marker PNCA were detected by Western Blot.Results Compared with NP460 cells,the expression of YBX3 was significantly increased in all nasopharyngeal carcinoma cells,the difference was statistically significant(P<0.01).The expression was the highest in 5-8 F cells and the difference was the most obvious(P<0.01).SiRNA and lentivirus ShRNA were used to interfere with 5-8 F cells of nasopharyngeal carcinoma.The interference efficiency of YBX3 was significantly reduced,and the difference was statistically significant(P<0.01).The results of proliferation experiments including CFSE,CCK8 and Clonal formation shown that interference with YBX3 significantly reduced cell proliferation,and the difference was statistically significant(P<0.01).The tumor-bearing experimental results in vivo also shown that the tumor growth in YBX3-ShRNA group was significantly reduced and the difference was statistically significant(P<0.01).In addition,the activity of MAPK p38 pathway and the expression of downstream key protein marker PCNA were significantly inhibited after using Lentivirus ShRNA interference experiment.Conclusion The results shown that downregulation the expression of YBX3 can inhibit cell proliferation and tumor growth of nasopharyngeal carcinoma through MAPK p38 pathway.