Fat1通过细胞外调节蛋白激酶信号通路抑制食管鳞癌细胞增殖  

Fat1 inhibits cell proliferation via ERK signaling pathway in esophageal squamous cell carcinoma

作　　者：杨斌[1] 张云魁[1] 叶永菁[1] 刘陶东[1] 彭胜祖[1] 张荣生[1] Yang Bin;Zhang Yunkui;Ye Yongjing;Liu Taodong;Peng Shengzu;Zhang Rongsheng(The Third Department of Thoracic Surgery,Shanxi Cancer Hospital,Taiyuan 030013,China)

机构地区：[1]山西省肿瘤医院胸外三病区,太原030013

出　　处：《中华肿瘤杂志》2021年第5期523-527,共5页Chinese Journal of Oncology

基　　金：山西省自然科学基金(201701D121166)。

摘　　要：目的探讨Fat1对食管鳞癌(ESCC)细胞增殖能力的影响及其机制。方法以Plko.1-puro-GFP-shRNA-Fat1质粒转染KYSE450细胞,应用实时定量PCR检测Fat1敲低效率。采用四甲基偶氮唑蓝法检测Fat1和细胞外调节蛋白激酶(ERK)通路抑制剂U0126对KYSE450细胞增殖能力的影响,平板克隆实验观察Fat1对ESCC细胞克隆形成能力的影响,活细胞动态观察细胞周期,Western blot方法检测相关靶蛋白的表达。通过裸鼠体内成瘤实验检测KYSE450细胞敲低Fat1后对移植瘤生长的影响,免疫组化方法检测肿瘤组织中相关蛋白的表达。结果Fat1sh1组和Fat1sh2组细胞中Fat1敲低效率分别为(77.1±6.9)%和(77.7±7.1)%。与对照组相比,Fat1sh1组和Fat1sh2组细胞增殖明显加快(P<0.05),细胞中p-ERK1/2的表达明显增加。经U0126处理后,Fat1敲低促进KYSE450细胞增殖的作用消失,细胞中p-ERK1/2的表达下降到与对照组相近的水平。对照组的细胞克隆数为(72±8)个,Fat1sh1组和Fat1sh2组分别为(155±28)个和(193±9)个,均高于对照组(均P<0.05)。对照组细胞分裂1次的时间为(1622±32)min,Fat1sh1组细胞分裂1次的时间为(1408±29)min,比对照组明显缩短(P<0.05)。裸鼠成瘤实验显示,Fat1敲低组比对照组裸鼠移植瘤增长速度快,接种后4周,对照组和Fat1敲低组瘤体重量分别为(0.224±0.028)g和(1.532±0.196)g,差异有统计学意义(P<0.05)。结论Fat1可通过抑制ERK信号通路进而抑制ESCC细胞增殖。Objective To clarify the mechanism of Fat1 on the proliferation of esophageal squamous cell carcinoma(ESCC).Methods KYSE450 cells were transfected with Plko.1-puro-GFP-shRNA-Fat1 plasmid and real time polymerase chain reaction(PCR)was used to verify the efficiency of Fat1 knockdown.The effects of Fat1 and extracellular regulated protein kinase(ERK)pathway inhibitor U0126 on the proliferation of ESCC cells were detected by methyl thiazolyl tetrazolium(MTT).Colony formation assay was used to detect the colony formation ability.Cell cycle was detected by live cell imaging.Western blot was used to observe the level of target protein.Mouse xenograft assay was applied to detect the effect of Fat1 knockdown on KYSE450 cell tumor growth.Immunohistochemistry was used to detect the expressions of related proteins in tumor sections.Results The efficiency of Fat1 knockdown was(77.1±6.9)%in Fat1 sh1 group and(77.7±7.1)%in Fat1sh2 group.Compared with the control group,the cell proliferation and the expression of p-ERK1/2 were significantly increased in Fat1 sh1 and Fat1sh2 group(P<0.05).After U0126 treatment,the effect of Fat1 knockdown on the proliferation of KYSE450 cells disappeared,and the expression of p-ERK1/2 in KYSE450 cells decreased to a level similar to that in the control group.The number of cell clones in the control group was(72±8),lower than(155±28)and(193±9)in the Fat1sh1 and Fat1sh2 groups,respectively(P<0.05).In KYSE450 cell,division time was shortened from 1622±32 min in control group to 1408±29 min in Fat1 sh1 group,the difference was statistically significant(P<0.05).Compared with the control group,the tumor volume of Fat1 knockdown group increased significantly.The tumor weight of control group and Fat1 knockdown group were(0.224±0.028)g and(1.532±0.196)g,respectively,at 4 weeks after inoculation,and the difference was statistically significant(P<0.05).Conclusion Fat1 inhibits cell proliferation via ERK signaling in ESCC.

关 键 词：食管鳞癌 Fat1 细胞增殖 细胞外调节蛋白激酶信号通路 

分 类 号：R735.1[医药卫生—肿瘤]