【摘要】：目的:肝細(xì)胞癌(Hepatocellular carcinoma,HCC)是全球常見(jiàn)惡性腫瘤,位列我國(guó)惡性腫瘤死因第2位。Wnt信號(hào)通路(Wnt signaling pathway)是一條在進(jìn)化上極其保守的信號(hào)傳導(dǎo)途徑,在胚胎發(fā)育、組織再生、造血、細(xì)胞增殖分化與遷移等生物學(xué)過(guò)程和功能中均發(fā)揮了十分重要的作用。此外,有研究顯示W(wǎng)nt信號(hào)通路在許多組織和器官的干細(xì)胞更新和分化中起介導(dǎo)作用。Wnt信號(hào)通路的異常激活已被證實(shí)與包括HCC在內(nèi)多種腫瘤發(fā)生發(fā)展密切相關(guān)。HCC是我國(guó)特色腫瘤,但我們目前對(duì)國(guó)人HCC中Wnt信號(hào)通路基因表達(dá)情況仍不明了。本研究旨在篩選國(guó)人HCC中Wnt信號(hào)通路差異表達(dá)基因,探討Wnt信號(hào)傳導(dǎo)通路差異表達(dá)基因在國(guó)人HCC發(fā)生發(fā)展過(guò)程中的作用,為尋找HCC潛在分子治療靶點(diǎn)提供依據(jù)。方法:本研究應(yīng)用Affymetrix U133 Plus2.0基因芯片檢測(cè)93例HCC及其配對(duì)癌旁正常肝組織中78個(gè)Wnt信號(hào)傳導(dǎo)通路相關(guān)基因的mRNA表達(dá)水平,篩選差異表達(dá)基因。HCC與配對(duì)癌旁正常肝組織間各基因表達(dá)水平比較采用配對(duì)t檢驗(yàn)。差異表達(dá)基因篩選標(biāo)準(zhǔn):t檢驗(yàn)P0.001,且癌較癌旁肝組織基因表達(dá)上調(diào)2倍(表達(dá)上調(diào)者)或下調(diào)50%(表達(dá)下調(diào)者)。癌組織中差異基因表達(dá)水平間的相關(guān)性采用Spearman相關(guān)檢驗(yàn)。差異表達(dá)基因的表達(dá)水平與臨床病理學(xué)參數(shù)的相關(guān)性分析采用χ2檢驗(yàn)。采用Stata 13.0軟件進(jìn)行統(tǒng)計(jì)分析。結(jié)果:1.與癌旁正常肝組織相比,HCC中TCF19、MMP12、DKK1、BCL9、SFRP4、MMP1、PYGO2、FZD6、MMP9、WNT6等10個(gè)基因表達(dá)上調(diào),而SFRP1、TCF21、SFRP5、FZD1、WNT2等5個(gè)基因表達(dá)下調(diào)。其中,TCF19表達(dá)上調(diào)倍數(shù)最多,達(dá)9.61倍(P=1.07×10-26);2.15個(gè)HCC差異表達(dá)基因的表達(dá)水平間存在廣泛相關(guān)性。TCF19表達(dá)與WNT6、FZD1、FZD6、MMP1、MMP12和BCL9表達(dá)水平均呈顯著正相關(guān)(均P0.05)。Wnt拮抗家族SFRPs和DKK中差異表達(dá)基因間的相關(guān)性:DKK1與SFRP4、SFRP4與SFRP1、SFRP1與SFRP5以及SFRP4與SFRP5的表達(dá)水平間均呈顯著正相關(guān)(均P0.05)。而DKK1與SFRP1及SFRP5的表達(dá)水平間均無(wú)顯著相關(guān)性(均P0.05);3.TCF19高表達(dá)(≥中位表達(dá)水平)與HCC多發(fā)(P=0.017)和低分化(P=0.046)顯著相關(guān);DKK1高表達(dá)(≥中位表達(dá)水平)與高AFP(20μg/L,P=0.026)、血管侵犯(P0.001)及低分化(P0.001)顯著相關(guān);SFRP4高表達(dá)(≥中位表達(dá)水平)與高AFP(P=0.008)和低分化(P=0.046)顯著相關(guān);SFRP1高表達(dá)與肝硬化(P=0.036)和TNM分期(P=0.033)顯著相關(guān);SFRP5高表達(dá)與AFP(P=0.009)、血管侵犯(P=0.011)、Edmondson-Steiner分級(jí)(P=0.022)及TNM分期(P=0.024)顯著相關(guān)。結(jié)論:Wnt信號(hào)傳導(dǎo)通路中多個(gè)基因異常表達(dá)參與了HCC的演進(jìn),研究結(jié)果支持Wnt信號(hào)傳導(dǎo)通路異常在我國(guó)HCC演進(jìn)過(guò)程中發(fā)揮重要作用,為Wnt信號(hào)通路在國(guó)人HCC中的作用機(jī)制研究提供了基礎(chǔ)。核內(nèi)轉(zhuǎn)錄因子TCF19是其中表達(dá)上調(diào)倍數(shù)最多的基因,可能體現(xiàn)了Wnt信號(hào)通路激活的關(guān)鍵終末效應(yīng),因而可能成為HCC分子治療的潛在靶點(diǎn),故其在HCC中的生物學(xué)及病理學(xué)意義有待進(jìn)一步研究。
[Abstract]:Objective: hepatocellular carcinoma (Hepatocellular carcinoma,HCC) is a common malignant tumor in the world. It ranks second in the cause of death of malignant tumors in China. Wnt signaling pathway (Wnt signaling pathway) is an evolutionarily conserved signal transduction pathway, which can be used in embryonic development, tissue regeneration and hematopoiesis. Cell proliferation, differentiation, migration and other biological processes and functions play a very important role. In addition, some studies have shown that Wnt signaling pathway mediates the regeneration and differentiation of stem cells in many tissues and organs. The abnormal activation of Wnt signaling pathway has been proved to be closely related to the occurrence and development of many tumors, including HCC. However, we still do not understand the gene expression of Wnt signaling pathway in Chinese HCC. The purpose of this study was to screen differentially expressed genes of Wnt signaling pathway in Chinese HCC and to explore the role of differential expression genes of Wnt signal transduction pathway in the pathogenesis and development of Chinese HCC. Methods: Affymetrix U133 Plus2.0 gene chip was used to detect the mRNA expression of 78 Wnt signal transduction pathway related genes in 93 cases of HCC and their matched normal liver tissues. The gene expression levels of HCC and matched normal liver tissues were compared by paired t test. The differential expression gene screening criteria were: P0.001, and the gene expression of cancer was increased by 2 times (the expression was up-regulated) or by 50% (the expression was down-regulated) than that of the adjacent liver tissue. The correlation of differentially expressed genes in cancer tissues was examined by Spearman correlation test. The correlation between the expression level of differentially expressed genes and clinicopathological parameters was analyzed by 蠂 2 test. Stata 13.0 software was used for statistical analysis. The result is 1: 1. Compared with the adjacent normal liver tissues, 10 TCF19,MMP12,DKK1,BCL9,SFRP4,MMP1,PYGO2,FZD6,MMP9,WNT6 genes were up-regulated and 5 SFRP1,TCF21,SFRP5,FZD1,WNT2 genes were down-regulated. The upregulation of TCF19 expression was the most. 2.15 HCC differentially expressed genes were widely correlated. TCF19 expression was significantly positively correlated with WNT6,FZD1,FZD6,MMP1,MMP12 and BCL9 expression levels (P0.05). Wnt antagonistic family SFRPs and DKK differentially expressed genes in the correlation between DKK1 and SFRP4,SFRP4 and SFRP1,SFRP1 There was a significant positive correlation with the expression of SFRP5, SFRP4 and SFRP5 (P 0.05). However, there was no significant correlation between the expression level of DKK1 and SFRP1 and SFRP5 (P0.05). 3. The high expression of TCF19 (鈮，

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