Y-Box结合蛋白1/锌指蛋白转录因子5轴对前列腺癌细胞成瘤性的影响及其机制

Effect of Y-Box binding protein-1/zinc finger protein transcription factor 5 axis on the tumorigenicity of prostate cancer cells and mechanism

目的探讨Y-Box结合蛋白1(YB-1)通过调控锌指蛋白转录因子5(KLF5)对前列腺癌细胞成瘤能力的影响。方法采用对照和YB-1短发卡RNA(shRNA)慢病毒感染感染人前列腺癌细胞株LNCaP,构建对照组和YB-1 KD组细胞,采用细胞计数试剂盒(CCK-8),EdU染色、克隆形成实验和体内移植瘤实验分析两组细胞的增殖和生长能力。采用荧光定量和蛋白质印迹法(Western blot)分析YB-1的靶基因KLF5 mRNA和蛋白表达水平。组间计量数据比较采用t检验。结果对照组细胞吸光度(A)值(2.04±0.11)明显高于YB-1 KD组(1.36±0.11),差异有统计学意义(t=10.930,P<0.05)。对照组细胞EdU阳性率[(85.41±4.13)%]明显高于YB-1 KD组[(63.34±5.70)%],差异有统计学意义(t=7.680,P<0.05)。对照组细胞克隆形成率[(76.72±5.51)%]明显高于YB-1 KD组[(53.30±4.93)%],差异有统计学意义(t=7.759,P<0.05)。对照组体内成瘤体积[(806.90±60.26)mm3]明显高于YB-1 KD组[(507.39±64.94)mm3],差异有统计学意义(t=10.690,P<0.05)。对照组成瘤重量[(7.01±0.92)g]明显高于YB-1 KD组[(4.80±0.73)g],差异有统计学意义(t=5.942,P<0.05)。对照组细胞KLF5 mRNA和蛋白表达水平(1.27±0.05、1.08±0.14)明显高于YB-1 KD组(0.56±0.11、0.53±0.09),差异有统计学意义(t=14.100、8.191,P<0.05)。结论YB-1蛋白参与前列腺癌细胞的增殖和成瘤效应,主要通过调节KLF5的表达水平来实现。

Objective To investigate the effect of Y-Box binding protein-1(YB-1)on the tumorigenic ability of prostate cancer cells by regulating zinc finger protein transcription factor 5(KLF5).Methods The human prostate cancer cell line LNCaP was infected with control and YB-1 short hairpin RNA(shRNA)lentivirus,and the control group and YB-1 KD group cells were constructed.The proliferation and growth abilities of the two groups of cells were analyzed using cell counting kit-8(CCK-8),EdU staining,clone formation experiments,and in vivo tumor transplantation experiments.The KLF5 mRNA and protein expression levels were analyzed by fluorescence quantification and Western blotting.The comparison of measurement data between groups was conducted using t-test.Results The absorbance(A)value in the control group(2.04±0.11)was significantly higher than that in the YB-1 KD group(1.36±0.11,t=10.930,P<0.05).The EdU positive rate in the control group[(85.41±4.13)%]was significantly higher than that in the YB-1 KD group[(63.34±5.70)%,t=7.680,P<0.05].The cell clone formation rate in the control group[(76.72±5.51)%]was significantly higher than that in the YB-1 KD group[(53.30±4.93)%,t=7.759,P<0.05].The tumor formation volume in the control group[(806.90±60.26)mm3]was significantly greater than that in the YB-1 KD group[(507.39±64.94)mm3,t=10.690,P<0.05].The tumor weight in the control group[(7.01±0.92)g]was significantly greater than that in the YB-1 KD group[(4.80±0.73)g,t=5.942,P<0.05].The expression levels of KLF5 mRNA and protein in the control group(1.27±0.05,1.08±0.14)were significantly higher than those in the YB-1 KD group(0.56±0.11,0.53±0.09),with statistically significant differences(t=14.100,8.191,P<0.05).Conclusion YB-1 protein participates in the proliferation and tumorigenesis of prostate cancer cells,mainly by regulating the expression level of KLF5.