AF1q在食管癌中的表达及其对细胞增殖和转移的影响

Expression of AF1q in esophageal carcinoma and its effect on cell proliferation and metastasis

目的探讨AF1q在食管癌中的表达及对细胞增殖和转移的影响。方法选取2018年1月到2022年1月商丘市第一人民医院收集的47例食管癌和癌旁组织作为研究对象,采用荧光定量聚合酶链反应(PCR)和蛋白质免疫印迹分析食管癌和癌旁组织AF1q mRNA和蛋白质表达水平;在人食管癌细胞株TE-1细胞采用慢病毒感染建立对照组和AF1q KD组稳定细胞系,采用细胞计数试剂盒(CCK-8)、体外移植瘤、划痕实验、Transwell分析对照组和AF1q KD组细胞增殖、迁移和侵袭能力。采用荧光定量PCR分析AF1q下游基因的表达水平。组间计量数据比较采用t检验。结果癌旁组织中AF1q mRNA表达水平(1.05±0.18)明显低于食管癌组织AF1q mRNA表达水平(2.10±0.17),差异有统计学意义(t=28.960,P<0.05)。蛋白质印迹法(Western blot)结果显示,癌旁组织中AF1q蛋白表达水平(0.75±0.10)明显低于食管癌组织AF1q蛋白表达水平(1.52±0.21),差异有统计学意义(t=23.460,P<0.05)。对照组细胞48 h吸光度值(2.03±0.11)明显高于AF1q KD组细胞(1.39±0.09),差异有统计学意义(t=10.900,P<0.05)。对照组细胞裸鼠体内30 d肿瘤体积[(806.83±47.88)mm^(3)]明显高于AF1q KD组细胞[(529.83±34.37)mm^(3)],差异有统计学意义(t=11.510,P<0.05)。对照组细胞裸鼠体内30 d肿瘤重量[(3.37±0.41)g]明显高于AF1q KD组细胞[(2.24±0.22)g],差异有统计学意义(t=5.918,P<0.05)。对照组细胞划痕愈合率[(85.83±4.71)%]明显高于AF1q KD组细胞[(65.83±7.36)%],差异有统计学意义(t=5.607,P<0.05)。Transwell结果显示,对照组细胞侵袭数量[(150.50±18.49)个]明显高于AF1q KD组细胞[(86.33±6.44)个],差异有统计学意义(t=8.027,P<0.05)。对照组细胞Wnt1、β-catenin和CD44蛋白表达水平(1.43±0.12、1.17±0.09、1.21±0.09)明显高于AF1q KD组细胞(1.06±0.05、0.70±0.09、0.85±0.05),差异有统计学意义(t=6.950、8.287、8.158,P<0.05)。结论AF1q在食管癌中呈高表达,通过调控Wnt信号通路参与食管癌细胞增殖,迁移和侵袭等过程。

Objective To investigate the expression of AF1q in esophageal carcinoma and its effect on cell proliferation and metastasis.Methods 47 cases of esophageal cancer and adjacent tissues collected by our hospital from January 2018 to January 2022 were selected as research objects.The expression levels of AF1q mRNA and protein in esophageal cancer and adjacent tissues were analyzed by fluorescent quantitative polymerase chain reaction(PCR)and Western blotting.In human esophageal cancer cell line TE-1,a stable cell line was established in the control group and AF1q KD group by lentivirus infection.The cell proliferation,migration and invasion ability of the control group and AF1q KD group were analyzed by cell counting kit-8(CCK-8),tumor transplantation in vitro,scratch test and Transwell.The expression level of AF1q downstream proteins were analyzed by Western blotting.Results The expression level of AF1q mRNA in the adjacent tissues(1.05±0.18)was significantly lower than that in esophageal carcinoma(2.10±0.17,t=28.960,P<0.05).The expression level of AF1q protein in the adjacent tissues(0.75±0.10)was significantly lower than that in esophageal cancer tissues(1.52±0.21,t=23.460,P<0.05).The 48 h absorbance absorbance(A)value(2.03±0.11)of cells in the control group was significantly higher than that of cells in the AF1q KD group(1.39±0.09,t=10.900,P<0.05).The tumor volume of control group cells[(806.83±47.88)mm^(3)]in nude mice at 30 days was significantly higher than that of AF1q KD group[(529.83±34.37)mm^(3),t=11.510,P<0.05].The tumor weight of control group[(3.37±0.41)g]in nude mice for 30 days was significantly higher than that of AF1q KD group cells[(2.24±0.22)g,t=5.918,P<0.05].The scratch healing rate of cells in the control group[(85.83±4.71)%]was significantly higher than that of cells in the AF1q KD group[(65.83±7.36)%,t=5.607,P<0.05].The number of cell invasion in the control group[(150.50±18.49)cells]was significantly higher than that in the AF1q KD group[(86.33±6.44)cells,t=8.027,P<0.05].The expression level of Wnt1,β-catenin and CD44 protein in control group cells(1.43±0.12;1.17±0.09;1.21±0.09)was significantly higher than that of AF1q KD cells(1.06±0.05;0.70±0.09;0.85±0.05,t=6.950,8.287,8.158,P<0.05).Conclusion AF1q is highly expressed in esophageal cancer and participates in the proliferation,migration and invasion of esophageal cancer cells by regulating Wnt signaling pathway.