Complex Behavior of ALDH1A1 and IGFBP1 in Liver Metastasis from a Colorectal Cancer.
Complex Behavior of ALDH1A1 and IGFBP1 in Liver Metastasis from a Colorectal Cancer.
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ALDH1A1 和 IGFBP1 在结直肠癌肝转移中的复杂行为。
DOI：
10.1371/journal.pone.0155160
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Kim YS
中科院分区：
文献类型：
--
作者：
Kim JC;Ha YJ;Tak KH;Roh SA;Kim CW;Kim TW;Kim SK;Kim SY;Cho DH;Kim YS
利用我们先前通过RNA测序建立的数据集（GSE 50760），本研究旨在鉴定与结直肠癌（CRC）肝转移（CLM）相关的上调基因，并验证其生物学行为。使用细胞增殖和侵袭测定来评估候选基因在肿瘤中的潜在作用。从18名患有同步CLM的CRC患者收集组织样品，并使用两种CRC细胞系（SW 480和SW 620）进行转染和克隆。在48只裸鼠脾内移植CRC细胞后，使用免疫组化验证CLM中鉴定的基因的作用。mRNA和蛋白质表达分别通过实时定量逆转录聚合酶链反应和蛋白质印迹法测定。根据其分子功能和生物学过程的相关性初步选择了9个基因，最后根据mRNA表达差异和蛋白表达正相关性选择了ALDH 1A 1和IGFBP 1。在SW 480细胞系中，ALDH 1A 1和IGFBP 1的过表达显著且时间依赖性地降低了细胞增殖（p ≤ 0.001-0.003），并抑制了侵袭性，抑制程度是对照细胞的3倍以上（p < 0.001），而它们对降低SW 620细胞增殖具有轻微作用。CLM中E-cadherin、N-cadherin、claudin-1和vimentin的蛋白表达水平显著高于原发肿瘤组织（p < 0.05）。然而，在CLM组织和转染的CRC细胞中未观察到钙粘蛋白转换，即N-钙粘蛋白过表达和E-钙粘蛋白表达降低。尽管体外细胞分析发现增殖和侵袭减少，但IGFBP 1过表达的SW 480细胞中β-连环蛋白、波形蛋白和ZO-1的持续过表达可能有助于植入IGFBP 1过表达的SW 480细胞的小鼠中CLM的发展（CLM发生率：SW 480/IGFBP 1转染小鼠对比SW 480/载体和SW 480/ALDH 1A 1转染小鼠，4/8对比0/10，p = 0.023）。总之，ALDH 1A 1和IGFBP 1在CLM中差异过表达，并可能发挥双重作用，在CRC中既作为肿瘤抑制因子又作为转移促进因子。
Using our data set (GSE50760) previously established by RNA sequencing, the present study aimed to identify upregulated genes associated with colorectal cancer (CRC) liver metastasis (CLM) and verify their biological behavior. The potential roles of candidate genes in tumors were assessed using cell proliferation and invasion assays. Tissue samples were collected from 18 CRC patients with synchronous CLM and two CRC cell lines (SW480 and SW620) were used for transfection and cloning. The roles of the genes identified in CLM were verified using immunohistochemistry in 48 nude mice after intrasplenic transplantation of CRC cells. mRNA and protein expression was determined by quantitative real-time reverse transcription polymerase chain reaction and western blot, respectively. Nine genes were initially selected according to the relevance of their molecular function and biological process and, finally, ALDH1A1 and IGFBP1 were chosen based on differential mRNA expression and a positive correlation with protein expression. The overexpression of ALDH1A1 and IGFBP1 significantly and time-dependently decreased cell proliferation (p ≤ 0.001–0.003) and suppressed invasiveness by ≥3-fold over control cells (p < 0.001) in the SW480 cell line, whereas they had a slight effect on reducing SW620 cell proliferation. The protein expression levels of E-cadherin, N-cadherin, claudin-1, and vimentin were significantly higher in CLM than in primary tumor tissues (p < 0.05). However, the cadherin switch, namely, N-cadherin overexpression with reduced E-cadherin expression, was not observed in CLM tissues and transfected CRC cells. Irrespective of reduced proliferation and invasion found on in vitro cell assays, persistent overexpression of β-catenin, vimentin, and ZO-1 in IGFBP1-overexpressing SW480 cells possibly contributed to CLM development in mice implanted with IGFBP1-overexpressing SW480 cells (CLM occurrences: SW480/IGFBP1-transfected mice vs. SW480/vector- and SW480/ALDH1A1-transfected mice, 4/8 vs. 0/10, p = 0.023). In conclusion, ALDH1A1 and IGFBP1 are differentially overexpressed in CLM and may play a dual role, functioning as both tumor suppressors and metastasis promoters in CRC.