探讨SOAT1联合多种标记物在肝细胞性肝癌辅助诊断中的应用

目的:探究甾醇氧-酰基转移酶1(SOAT1)在肝细胞性肝癌组织中的表达情况,并探讨其联合多种标记物(HSP70、GS、CD10、CD34)在诊断肝细胞性肝癌中的应用可行性。方法:收集58例肝细胞性肝癌组织、配对癌旁肝组织及12例良性肝病变组织样本,制作组织芯片,免疫组织化学方法检测SOAT1、HSP70、GS、CD10、CD34的表达情况,并分析其表达相关性。结果:SOAT1在肝细胞性肝癌组织中表达上调(P<0.01),且表达水平与肝细胞性肝癌患者分化程度密切相关(P<0.05)。SOAT1与HSP70、GS、CD34表达相关(P<0.01);肝细胞性肝癌诊断阳性符合率分别为CD34100.00%、GS 89.66%、HSP7082.75%、CD1063.79%、SOAT163.79%;而在癌旁组织中诊断阴性符合率分别为SOAT1100.00%、CD1098.27%、HSP7096.55%、CD3434.48%、GS 5.17%;良性肝病变组织中诊断阴性符合率分别为SOAT1100.00%、CD1083.34%、HSP70100.00%、CD348.33%、GS 0%。以任意3种标记物阳性为诊断标准,不将SOAT1计入标记物行列,58例肝细胞性肝癌中有39例可被诊断,诊断率为67.24%;而将SOAT1计入标记物行列,58例肝细胞性肝癌中有48例可被诊断,诊断率可提高至82.76%。结论:SOAT1在肝细胞性肝癌中表达水平明显上调,且在低分化肝细胞性肝癌表达水平更高,提示SOAT1可作为肝细胞性肝癌的辅助诊断及预后判断指标之一。SOAT1、HSP70、GS、CD10及CD345种标记物中,敏感性较好为CD34及GS,而特异性较好则为SOAT1、CD10及HSP70。SOAT1与HSP70、GS、CD10、CD34联合应用,在肝细胞性肝癌的辅助诊断中有一定应用价值。

肝癌生物标志物SOAT1单克隆抗体的制备及评估

肝癌是临床上一种常见的恶性肿瘤,目前临床上仍缺乏特异有效的诊断标志物。有研究表明,固醇O-酰基转移酶1(sterol O-acyltransferase 1,SOAT1)作为胆固醇代谢的关键酶,与肝癌的发生发展密切相关。因此,SOAT1在作为肝癌生物标志物方面具有巨大潜力。本研究从肝癌细胞Huh-7中获取SOAT1基因,经原核表达、蛋白质纯化后,以纯化的重组SOAT1蛋白作为抗原免疫小鼠,利用杂交瘤等技术共获得5株SOAT1单克隆抗体细胞株,分别命名为1F3、1G3、1D6、2F8、4D11,其中4D11能够识别肝癌细胞Huh-7及临床肝癌组织标本中的SOAT1蛋白;最后,将4D11送至生物公司进行测序,获得该抗体可变区的氨基酸序列。本研究利用重组SOAT1蛋白成功制备了抗SOAT1的特异性单克隆抗体,这不仅为后续应用SOAT1蛋白制备肝癌伴随诊断试剂盒提供了原材料,而且还为建立基于SOAT1的肝癌诊断方法奠定了基础。

A pan-cancer analysis identifies SOAT1 as an immunological and prognostic biomarker

Sterol o-acyltransferase1(SOAT1)is an enzyme that regulates lipid metabolism.Nevertheless,the predictive value of SOAT1 regarding immune responses in cancer is not fully understood.Herein,we aimed to expound the predictive value and the potential biological functions of SOAT1 in pan-cancer.Raw data related to SOAT1 expression in 33 different types of cancer were acquired from The Cancer Genome Atlas(TCGA)and Genotype-Tissue Expression(GTEx)databases.SOAT1 expression was significantly increased in most cancers and showed a distinct correlation with prognosis.This enhanced expression of the SOAT1 gene was confirmed by evaluating SOAT1 protein expression using tissue microarrays.In addition,we found significant positive associations between SOAT1 expression levels and infiltrating immune cells,including T cells,neutrophils,and macrophages.Moreover,the co-expression analysis between SOAT1 and immune genes showed that many immune-related genes were increased with enhanced SOAT1 expression.A gene set enrichment analysis(GSEA)revealed that the expression of SOAT1 correlated with the tumor microenvironment,adaptive immune response,interferon signaling,and cytokine signaling.These findings indicate that SOAT1 is a potential candidate marker for predicting prognosis and a promising target for tumor immunotherapy in cancers.

Application of SOAT1 combined with multiple markers in the auxiliary diagnosis of hepatocellular carcinoma

Objective:To explore the expression of sterol oxygen-acyltransferase 1(SOAT1)in hepatocellular carcinoma tissues and to evaluate the application value of its combination with various markers(HSP70,GS,CD10,CD34)in the diagnosis of hepatocellular carcinoma.Methods:Totally 58 cases of liver cancer tissue and its corresponding adjacent tissue,and 12 cases of benign liver lesions tissue were collected,and tissue chips were made to detect the expressions of SOAT1,HSP70,GS,CD10 and CD34(immunohistochemical staining method)which were analyzed by scientific method.Results:The expression of SOAT1 was up-regulated in hepatocellular carcinoma tissues,and the difference was statistically significant(P<0.01),and the expression level was closely related to the differentiation degree of hepatocellular carcinoma patients(P<0.05).SOAT1 was correlated with the expression of HSP70,GS,and CD34,with statistical significance(P<0.01).Positive percent agreement for the hepatocellular carcinoma was 100.00% for CD34,89.66% for GS,82.75% for HSP70,63.79% for CD10,and 63.79% for SOAT1;negative percent agreement(paracancerous tissues)was 100% for SOAT1,98.27 %for CD10,96.55% for HSP70,34.48% for CD34,and 5.17% for GS;Negative percent agreement(benign tissues)was 100%for SOAT1,83.34% for CD10,100.00% for HSP70,8.33% for CD34,and 0% for GS,respectively.Taking the positive expression of any three markers as the diagnostic criteria,39 of the 58 cases of hepatocellular carcinoma could be diagnosed,and the diagnostic rate was 67.24%(SOAT1 was not included in the list of markers).If SOAT1 is included,48 of the 58 hepatocellular carcinoma cases could be diagnosed,and the diagnosis rate could be increased to 82.76%.Conclusions:The expression level of SOAT1 was significantly up-regulated in hepatocellular carcinoma,and the expression level was higher in poorly differentiated HCC,suggesting that SOAT1 can be used as one of the indicators for the diagnosis and prognosis of hepatocellular carcinoma.Among the five markers above,CD34 and GS were more sensitive,while the specificity is better for SOAT1,CD10 and HSP70.The combined application of SOAT1 with HSP70,GS,CD10 and CD34 has certain application value in the diagnosis of hepatocellular carcinoma.

新型SOAT1蛋白靶向抑制剂的筛选与体外活性考察

目的筛选新型甾醇O-酰基转移酶1(sterol O-acyltransferase,SOAT1)靶向抑制剂,并对其活性进行考察。方法通过CCLE数据库筛选SOAT1高表达肝癌细胞系,选择相应细胞系作为实验细胞模型。采用Autodock Vina软件对Interbioscreen数据库中61742个化合物与SOAT1蛋白进行分子对接,检测结合能。筛选结合力强的8个化合物,在细胞模型上用CCK8法检测其对肝癌细胞活力的影响。选择在细胞模型上活性最强的2个化合物进一步研究,并测定其IC50。划痕试验及结晶紫染色法进一步检测筛选出的2个化合物对肝癌细胞迁移和增殖的影响,并采用蛋白质免疫印迹法检测化合物对肝癌细胞中SOAT1蛋白的影响。转染siRNA构建SOAT1沉默的肝癌细胞模型,并检测化合物对肝癌细胞活力的影响。结果从CCLE数据库中筛选出SOAT1蛋白高表达的Hep3B和PLC/PRF/5肝癌细胞系作为模型细胞。通过分子对接筛选出的结合力前8的化合物中,化合物1和化合物7对上述2种肝癌细胞活力的抑制作用最为显著。此外,这2个化合物还能抑制肝癌细胞迁移及克隆形成,并降低肝癌细胞中SOAT1蛋白表达。在SOAT1沉默的肝癌细胞中,2个化合物对肝癌细胞活力的抑制作用均显著减弱。结论化合物1和化合物7在细胞水平上能通过抑制SOAT1蛋白表达发挥抗肝癌作用,有潜力开发为治疗肝癌的SOAT1抑制剂。

靶向胆固醇稳态小分子药物治疗胶质瘤的研究进展

胶质母细胞瘤(GBM)是成人中最具有侵袭性的恶性脑肿瘤。胆固醇是细胞的必要组分,胆固醇的合成、转运和代谢异常在肿瘤的发生发展过程中起着非常重要作用。胆固醇代谢稳态相关调控因子如尼曼-匹克样C型蛋白(NPC1)、甾醇O-酰基转移酶(SO⁃AT1)已成为肿瘤治疗中潜在的新药物干预靶点。该研究阐述了部分靶向胆固醇稳态的小分子药物可能对GBM的治疗作用。

纳米二氧化钛对小鼠卵巢组织中甾醇O-酰基转移酶1与类固醇合成急性调节蛋白表达的影响

目的研究纳米二氧化钛染毒对小鼠卵巢组织中甾醇O-酰基转移酶1(SOAT1)与类固醇合成急性调节蛋白(STAR)表达的影响。方法将48只40日龄清洁级ICR雌性小鼠随机分为4组,分别为对照(0.5%羧甲基纤维素)组和25、50、100 mg/kg纳米二氧化钛染毒组,每组12只。通过灌胃方式进行染毒,染毒容积为10 ml/kg,每天1次,连续染毒60 d。采用基因芯片检测100 mg/kg纳米二氧化钛染毒60 d小鼠卵巢组织中类固醇代谢相关基因表达的影响。采用实时定量PCR法检测100 mg/kg纳米二氧化钛染毒60 d后卵巢组织SOAT1与STAR基因表达的水平。使用ELISA法检测各剂量纳米二氧化钛染毒15、30、60 d后卵巢组织SOAT1与STAR蛋白水平。结果与对照组比较,以差异值≥13分或≤-13分为筛选标准,在100 mg/kg纳米二氧化钛染毒60 d小鼠卵巢中已知功能为类固醇代谢的差异表达基因中,Soat1、Star、Cyp17a1和Cyp11a1表达上调,Cyp2b1、Akr1c19和Cyp2a5表达下调。与对照组比较,100 mg/kg纳米二氧化钛染毒60 d小鼠卵巢中SOAT1与STAR基因实时定量PCR法的检测结果上调(P<0.01),与芯片结果一致。与对照组比较,50 mg/kg纳米二氧化钛染毒60 d和100 mg/kg纳米二氧化钛染毒30、60 d小鼠卵巢组织中STAR与STAR的蛋白水平上升,差异均有统计意义(P<0.05);且随着纳米二氧化钛染毒剂量的升高和染毒时间的延长,小鼠卵巢组织中SOAT1与STAR的水平呈现上升趋势。结论纳米二氧化钛暴露通过促进小鼠卵巢组织中SOAT1与STAR的表达,干扰卵巢类固醇代谢。

HBV-DNA、甾醇氧-酰基转移酶1表达及肝细胞癌分化程度的相关性

目的 探讨HBV-DNA水平、甾醇氧-酰基转移酶1(Sterol O-acyltransferase, SOAT1)表达及肝细胞癌肿瘤分化程度之间的相关性,为了解肝细胞癌发展提供科学依据。方法 收集58例HBV相关肝细胞癌病例及病历资料中HBV-DNA水平数据,同时收集病例癌组织及其配对癌旁组织,采用免疫组织化学方法检测SOAT1表达情况并评估肿瘤分化程度。结果 SOAT1高表达病例组和SOAT1低表达组HBV-DNA高水平率分别为81.1%(30/37)和19.0%(4/21),差异有统计学意义(χ^(2)=21.253,P<0.05),且SOAT1表达与HBV-DNA水平之间呈相关性(χ^(2)=7.021,P<0.05)。低分化肝细胞癌病例组中HBV-DNA高水平率为71.1%(27/38),中高分化肝细胞癌组中HBV-DNA高水平率为35.0%(7/20),差异具有统计学意义,且HBV-DNA水平与肿瘤分化程度之间呈现相关性(χ^(2)=7.021,P<0.05)。SOAT1在所有收集病例中整体高表达率为63.8%(37/58),在所有配对癌旁组织中均无表达(0/58),且SOAT1蛋白在癌组织中的表达水平与肿瘤分化程度呈现相关性(χ2=19.889,P<0.05)。在低分化病例组中SOAT1呈现高表达,高表达率为84.2%(32/38),在较高分化病例组的HCC样本中SOAT1普遍呈现低表达,高表达率仅为25.0%(5/20)。结论 HBV-DNA水平与肝细胞癌分化程度相关,低分化癌中HBV-DNA水平更高;SOAT1表达水平亦与肝细胞癌分化程度相关,低分化癌中SOAT1表达水平亦更高;且HBV-DNA水平与SOAT1表达水平呈正相关,而SOAT1是细胞脂质代谢的关键酶,提示HBV感染可能影响SOAT1的功能及水平从而干预肝细胞脂质代谢而参与肿瘤的发生及演进。

肝细胞癌相关固醇-O酰基转移酶1基因的生物信息学分析

目的使用生物信息学工具对肝细胞癌相关固醇-O酰基转移酶1(sterol O-acyltransferase 1,SOAT1)的结构、功能进行预测分析,以了解其作为S-Ⅲ亚型标记物以及治疗靶标的相关作用机制。方法使用NCBI、STRING、Protscale、SignalP、TMHMM、PSORT、SOPMA、SWISS-MODEL、NetNGlyc、NetOGlyc、Netphos、ProtParam等数据库或软件,对SOAT1的结构、功能、蛋白相互作用等进行预测分析。结果SOAT1编码的蛋白质为疏水性蛋白,稳定性较好,属于非经典途径蛋白,具有8个跨膜区域,主要分布于细胞膜。SOAT1在多种组织均有表达,但绝大多数位于肾上腺。具有多个磷酸化位点,主要参与胆固醇的合成以及分解代谢过程。结论通过生物信息学分析SOAT1的结构以及功能,发现SOAT1脂质的合成以及分解代谢途径发挥重要作用,脂质的表达与癌症的发展密切相关。因此可通过调节SOAT1基因的表达从而实现肝细胞癌的治疗。