| 夏明亮,任川川,李云龙,等.转录因子叉头框蛋白 O3a对前列腺癌细胞上皮间质转化的影响[J].安徽医药,2022,26(2):297-303. |
| 转录因子叉头框蛋白 O3a对前列腺癌细胞上皮间质转化的影响 |
| The effects of FOXO3a on epithelial-mesenchymal transition of prostate cancer cells |
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| DOI：10.3969/j.issn.1009-6469.2022.02.022 |
| 中文关键词: 前列腺肿瘤 上皮间质转化 转录因子叉头框蛋白 O3a Wnt/β-catenin信号通路 |
| 英文关键词: Prostatic neoplasms Epithelial-mesenchymal transition Transcription factor forkhead box protein O3a Wnt/β-catenin signaling pathway |
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| 中文摘要: |
| 目的探讨转录因子叉头框蛋白 O3a(FOXO3a)对前列腺癌细胞上皮间质转化( EMT)的影响及其作用机制。方法采用逆转录聚合酶链式反应( RT-PCR)检测正常前列腺上皮细胞 RWPE-1和前列腺癌细胞 LNCaP、DUl45、PC-3中 FOXO3a表达水平，选择 FOXO3a表达最低的前列腺癌细胞系转染 pcDNA-FOXO3a、pcDNA-NC构建 FOXO3a过表达细胞系( pc-FOXO3a)和阴性对照( NC)采用 MTT法、流式细胞法检测细胞活性和凋亡情况， Transwell试验、划痕试验检测细胞侵袭和迁移能力，白质印迹法(Wester，n blotting)检测钙黏附蛋白 E(E-cadherin)、波形蛋白( Vimentin)、神经型钙黏附蛋白( N-cadherin)、 Snail、c蛋-Myc、细胞周期蛋白 D1(cyclin D1)、剪切型胱天蛋白酶 -3(Cleaved-Cas-3)、 Bcl-2相关 X(Bax)、 B细胞淋巴瘤 -2(Bcl-2)、 β-连环蛋白( β-catenin)、 Wnt1水平。运用氯化锂( Wnt/β-catenin信号通路激活剂)作用于 pcDNA-FOXO3a前列腺细胞，分析细胞活性、凋亡和上皮间质转换( EMT)情况。结果 LNCaP、DUl45、PC-3细胞中 FOXO3a表达水平均显著低于 RWPE-1细胞［( 0.23± |
| 英文摘要: |
| Objective To explore the effects and action mechanism of forkhead box protein O3a (FOXO3a) on epithelial-mesenchy. mal transition (EMT) of prostate cancer cells.Methods The expression levels of FOXO3a in normal prostate epithelial cells (RWPE-1) and prostate cancer cells (LNCaP, DU145, PC-3) were detected by RT-PCR. The prostate cancer cell line with the lowest FOXO3a ex. pression was selected and was transfected with pcDNA-FOXO3a and pcDNA-NC to construct FOXO3a overexpression cell line (pc-FOXO3a) and negative control (NC). The cell viability and apoptosis were detected by MTT and flow cytometry. The invasion and migra.tion of cells were detected by Transwell assay and scratch assay. The levels of E-cadherin, Vimentin, N-cadherin, Snail, c-Myc, cyclin D1, Cleaved-Cas-3, Bax, Bcl-2, β-catenin and Wnt1 were detected by Western blot. The pcDNA-FOXO3a prostate cells were treated with lithium chloride (Wnt/β-catenin signaling pathway activator) to analyze cell viability, apoptosis and EMT.Results The expres. sion level of FOXO3a in LNCaP, DU145 and PC-3 cells was significantly lower than that in RWPE-1 cells [(0.23±0.04), (0.53±0.05), (0.42±0.06) vs. (1.02±0.08)] (P<0.05), and the expression level of FOXO3a in LNCaP cells was the lowest. The cells activity, number ofinvasion cells in per field and migration rate of scratches in pc-FOXO3a LNCaP cells were significantly lower than those in NC LNCaP cells (P<0.05), while apoptosis rate was significantly higher than that in NC LNCaP cells (P<0.05). The levels of N-cadherin, Vimentin, Snail, β-catenin, c-Myc, cyclin D1, Bcl-2 and Wnt1 in pc-FOXO3a LNCaP cells were significantly lower than those in NC LNCaP cells (P<0.05), while the levels of E-cadherin, Cleaved-Cas-3 and Bax were significantly higher than those in NC LNCaP cells (P<0.05). The cells activity, number of invasion cells in per field, migration rate of scratches, and levels of N-cadherin,Vimentin and Snail in LiCl-pc- |
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