肺鳞癌患者与健康人血清的差异蛋白质组学研究

为筛选肺鳞癌的血清标志物,采用二维凝胶电泳(2-DE)技术分离I期肺鳞癌患者和健康人的血清蛋白质,PDquest图像分析软件识别差异蛋白质点,电喷雾串联质谱(ESI-Q-TOF MS/MS)鉴定差异蛋白,然后应用蛋白质印迹和免疫组化方法分别检测差异蛋白——结合珠蛋白-2(haptoglobin-2,HP-2)在肺鳞癌患者血清和健康人血清以及肺鳞癌组织和癌旁正常支气管上皮组织中的表达.建立了肺鳞癌患者和健康人血清的2-DE图谱,图像分析软件识别了1O个差异蛋白质点,质谱鉴定了4种差异蛋白;蛋白质印迹分析显示,HP-2在肺鳞癌血清中的表达水平显著高于健康人(P<0.05),但其表达水平与肺鳞癌的临床分期无明显相关性;免疫组化结果显示,HP-2在肺鳞癌组织中的表达水平高于癌旁正常支气管上皮组织(P<0.05).研究结果提示:HP-2是候选的肺鳞癌血清分子标志物,血清中HP-2水平对肺鳞癌诊断可能具有一定的参考价值;肺鳞癌组织中HP-2表达上调可能是患者血清中HP-2表达升高的原因之一.     

肺癌病人血清中U1-A snRNP自身抗体的发现与鉴定

许多研究表明, 肺癌病人存在自身免疫现象. 为了寻找肺癌病人血清中具有潜在诊断价值的自身抗体, 采用免疫荧光染色, 免疫印迹和蛋白质芯片技术对10例肺癌病人和10例正常人血清进行了检测. 筛选中发现1例来自64岁男性肺鳞癌(Ⅲb期)病人的血清呈现特异性的细胞核染色, 并在31 kD处有一个明显的细胞核蛋白免疫印迹条带. 进而采用该病人的血清进行免疫沉淀, 对所捕获的蛋白组分进行质谱分析和数据库查询后确认31 kD条带为核内小核糖体蛋白U1-A(small nuclear ribonucleoprotein U1-A, U1-A snRNP). 进一步对36例鳞癌、26例腺癌、31例小细胞肺癌和20例健康对照血清样本进行了免疫荧光染色和免疫印迹验证, 结果显示50%的鳞癌、26.9%的腺癌、54.8%的小细胞肺癌病人血清中有抗U1-A snRNP抗体. 上述结果报道了在肺癌病人血清中出现Anti-U1-A snRNP自身抗体.     

人肺腺癌细胞A—549和正常细胞HBE的蛋白质组差异分析

为了研究人肺腺癌细胞A 5 49和正常细胞HBE的蛋白质组差异 ,用固相pH梯度双向凝胶电泳分离人肺腺癌细胞系A 5 49和正常细胞HBE的总蛋白质 ,银染显色 ,PDQuest 2 DE软件分析 ,对部分差异蛋白质点进行基质辅助激光解吸电离飞行时间质谱 (MALDI TOF MS)测定其胶内酶解后的肽质指纹图谱 ,用PeptIdent软件查询SWISS PROT数据库。结果获得了分辨率和重复性均较好的双向电泳银染图谱 ,图象分析探测到A 5 492 DE图谱的平均蛋白质点数为 (890± 38)个 ,HBE的平均蛋白质点数为 (75 7± 2 7)个 ,不同胶间蛋白质点的位置偏差在IEF方向为 (2 .85± 0 .48)mm ,在SDS PAGE方向为 (2 .6 9± 0 .37)mm。差异表达分析发现A 5 49和HBE图谱有5 35个蛋白质点相互匹配 ,其中A 5 49有 35 5个未被匹配 ,HBE中有 2 2 2个未被匹配 ;对A 5 49和HBE中的 18个差异蛋白质点分别进行肽质指纹分析 ,经数据库查询 ,初步鉴定为一些与物质代谢、细胞因子、信号转导有关的蛋白质。提示人肺腺癌细胞A 5 49和正常细胞HBE的蛋白质组具有差异 ,这种蛋白质组的差异分析有助于进一步研究肺腺癌的相关蛋白质及分子标记物     

应用二维电泳和质谱技术筛选喉癌及癌旁组织中的差异表达蛋白质

目的分离并鉴定喉癌和癌旁正常粘膜组织的差异表达蛋白质,为喉癌早期临床诊断、治疗提供新的有关的肿瘤生物学标记和靶标。方法收集5对人喉癌组织和对应的癌旁正常粘膜组织,提取组织总蛋白质,采用二维凝胶电泳技术分离蛋白并进行比较。选择在喉癌中明显差异表达的蛋白质点,进行质谱分析。结果获得了分辨率和重复性均较好的凝胶蛋白图谱。筛选出的在喉癌及癌旁正常粘膜组织中明显差异表达的10个蛋白质点,并成功鉴定。其中在喉癌组织中高表达的7个,低表达的3个。结论喉癌组织与癌旁正常粘膜组织蛋白存在明显的差异,筛选并鉴定出的这些蛋白质可能成为喉癌早期临床诊断、治疗的标志物和靶标。     

一种提高固定pH梯度（IPG）凝胶双向电泳的重复性、分辨率和通量的方法

在蛋白质组学研究中 ,双向聚丙烯酰胺凝胶电泳是现行蛋白质分离的最重要的方法之一。实验发展了一种提高固定pH梯度 (IPG)凝胶双向电泳的重复性、分辨率和通量的方法 :在一块SDS 聚丙烯酰胺凝胶上同时进行多块固定pH梯度(IPG)凝胶 (Multi stripsononeSDSgel,MSOG)电泳。用此方法比较了人肝癌细胞、不同生长状态的人肝癌细胞、3T3细胞的蛋白质以及同一个样品在不同大小的第二向凝胶系统 (大型和中型凝胶 )的双向电泳图谱。结果表明 ,同一样品在 13cmIPGStrip双向电泳可分离 2 0 0 0以上蛋白质点且图谱蛋白质点的匹配率可超过 95 %以上。同时又可以最大程度地降低凝胶背景对蛋白质点比较分析的干扰 ,从而提高了双向电泳分离蛋白质的分辨率和通量。这些优点都有助于差异蛋白质组学特别是细胞器差异蛋白质组学研究的自动化。     

激光捕获显微切割技术纯化的鼻咽癌和正常鼻咽上皮细胞的比较蛋白质组学研究

为筛选鼻咽癌(nasopharyngeal carcinoma, NPC)的分子标志物,采用激光捕获显微切割技术(laser capture microdissection, LCM)分别从NPC组织和正常鼻咽上皮组织(normal nasopharyngal epithelial tissue, NNET)中切割纯化NPC细胞和正常鼻咽上皮细胞(normal nasopharyngal epithelial cells, NNEC),应用二维凝胶电泳(two-dimensional electrophoresis, 2-DE)分离LCM纯化细胞的蛋白质,图像分析识别差异表达的蛋白质点,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)和电喷雾电离串联质谱(ESI-Q-TOF-MS)鉴定差异蛋白质点,Western blot检测差异蛋白cytokeratin 8 (CK8)在LCM纯化的NPC细胞和NNEC以及具有不同分化程度或转移潜能的4株NPC细胞中的表达,免疫组织化学检测CK8在63例NPC、28例NNET及20例颈淋巴结转移NPC组织中的表达水平.建立了LCM纯化的NPC细胞和NNEC的2-DE图谱,质谱鉴定了29个差异蛋白质,其中15个蛋白质只在NPC表达或表达明显增高,14个蛋白质在NPC中表达下调或缺失;Western blot结果显示,CK8的表达水平在NPC中较NNET明显下调,并与NPC细胞株的分化程度和转移潜能有关;免疫组织化学结果显示,CK8在NPC组织中的表达较NNET明显下调,在颈淋巴结转移NPC中的表达较原发NPC明显上调.研究结果提示,CK8与NPC分化及淋巴结转移相关,有望成为预测NPC转移和区别NPC分化程度的分子标志物.     

LCM纯化的人支气管上皮癌变各阶段组织的定量蛋白质组学研究

为分析支气管上皮癌变进程中的差异表达蛋白质,筛选肺鳞癌早期诊断标志物,以人支气管上皮癌变各阶段组织为研究对象,先采用激光捕获显微切割技术(LCM) 纯化人正常支气管上皮组织、鳞状化生、不典型增生、原位癌、浸润性肺鳞癌组织,再用同位素标记相对和绝对定量 (iTRAQ) 技术结合二维液相色谱串联质谱（2D LC-MS/MS）鉴定支气管上皮癌变进程中各阶段的差异表达蛋白质。结果共鉴定了1036个蛋白质,筛选出102个与人支气管上皮癌变相关的差异蛋白质,在这些差异蛋白质中,有的在支气管上皮癌变过程中进行性上调,有的在支气管上皮癌变过程中进行性下调,有的呈阶段特异性改变;功能分析表明,这些差异蛋白质涉及代谢、细胞凋亡、增殖、分化、信号传导、转录、翻译、细胞粘附、免疫反应与发育等。Western blotting 及免疫组织化学技术验证了其中 2个差异蛋白(S100A9和 CKB) 的表达,证实了定量蛋白质组学结果的可靠性。研究结果提示：这些差异表达蛋白质与支气管上皮癌变相关,并可成为肺鳞癌的早期诊断标志物,进一步研究差异蛋白的生物学功能,将有助于阐明支气管上皮的癌变机制,从而为肺鳞癌的早期诊断与发病机制研究提供新思路。     

应用蛋白质组学的方法筛选胰腺干细胞标记物

探索利用蛋白质组学的方法筛选胰腺干细胞的标记物.通过大鼠胰腺大部分切除(Px)的方法建立胰腺增生模型,采用双向电泳(2DE)分离比较Px术后第3天大鼠增生胰腺组织与假手术(Sx)非增生胰腺组织蛋白质的表达谱差异,用基质辅助激光解吸离子化飞行时间质谱(MALDI-TOF-MS)和肽质量指纹图谱(PMF)的方法对差异蛋白进行鉴定,进一步经生物信息学检索,筛选与胚胎发育和细胞分化相关的蛋白质分子.蛋白质印迹分析蛋白质HSP47、Vimentin在Px大鼠胰腺组织中的表达,以验证2DE的结果.2DE显示Sx和Px大鼠胰腺组织平均蛋白质点数分别为(1369±31)和(1315±28)个,其中共有91个蛋白质点出现1.5倍以上的表达差异.所有差异蛋白质点PMF鉴定出53种蛋白质,其中Vimentin,cytokeratin8(CK8),lymphocytecytosolicprotein1(L-plastin),heterogeneousnuclearribonucleoproteinA2/B1(hnRNPA2/B1)和L-arginine:glycineamidinotransferase(AGAT)与胚胎发育和细胞分化有关,这些蛋白质可能是胰腺干细胞潜在的候选标记物.总之,蛋白质组学的方法利用其高通量的特点能有效发现潜在的胰腺干细胞生物学标记物.     

长链非编码RNA SNHG17通过抑制p57表达促进肺癌进展

目的:分析肺癌中长链非编码RNA(lncRNA)小核仁RNA宿主基因17(SNHG17)与p57的表达相关性及功能联系。方法:通过starBase数据库分析SNHG17和p57在肺癌组织和正常组织中的表达差异;采用人正常肺上皮细胞BEAS-2B和人肺癌细胞系A549、H1975进一步分析SNHG17和p57在肺癌细胞和正常肺细胞中的表达差异;采用qRT-PCR和Western印迹检测si-SNHG17在mRNA和蛋白表达方面对SNHG17和p57的影响;采用CCK-8法检测siSNHG17对A549细胞增殖的影响;采用Transwell法检测si-SNHG17对A549细胞迁移和侵袭能力的影响。结果:通过数据分析发现SNHG17在肺腺癌和肺鳞癌组织中显著高表达,而p57在肺腺癌和肺鳞癌组织中却显著低表达;与正常肺细胞相比,SNHG17和p57在肺癌细胞中的表达趋势与在肺癌组织中一致;SNHG17和p57的表达呈负相关;沉默SNHG17使p57的表达一定程度上调,A549细胞的增殖、迁移和侵袭能力受到抑制。结论:SNHG17在肺癌中高表达,促进肺癌细胞增殖,并通过调节p57的表达来抑制肺癌细胞的增殖和迁移。SNHG17和p57在肺癌中的具体作用机制仍有待进一步研究。     

胆管癌患者与健康志愿者血清样品蛋白质表达差异分析

胆管癌（cholangiocarcinoma,CCA）是一种难于早期诊断和治疗的恶性肿瘤.利用蛋白质组学技术以期筛选出人血清中具有医学价值的胆管癌癌变的标志物.利用白蛋白/IgG去除试剂盒去除血清中高丰度的白蛋白和IgG,再经丙酮沉淀得到高质量的血清样品.样品经双向电泳（2-DE）得到了分辨率较好的胆管癌血清的蛋白点图谱.利用Image Master 2D软件对比分析了胆管癌病人和正常健康对照组的2-DE图谱.其中,表达上调蛋白有8个,表达下调蛋白有6个.用质谱获得上述差异蛋白的肽质量指纹图谱,并经数据库检索共鉴定出了11种差 异蛋白质,其中8种蛋白质的表达与胆管癌密切相关.本研究为阐明胆管癌的机理提供了一条新途径.     

Proteome analysis of human lung squamous carcinoma

Few lung cancer-specific molecular markers have been established in regard of "early-stage" diagnosis and prognosis. In this study the proteome analysis of human lung squamous carcinoma (hLSC) was carried out using two strategies to explore the carcinogenic mechanisms and identify its molecular markers more directly and comprehensively. Comparative proteome analysis on 20 hLSC tissues and paired normal bronchial epithelial tissues revealed 76 differential proteins, among which 68 proteins were identified by PMF. The identified proteins fell into three categories: oncoproteins, cell cycle regulators and signaling molecules. To validate the identified differential proteins, the expressions levels of three differential proteins mdm2, c-jun and EGFR were determined by immunohistochemical staining and immunoblots. The results verified proteome analysis results. Serological proteome analysis (SERPA) of ten hLSC tissues was performed to identify the tumor-associated antigens. The results revealed 36 +/- 8 differential proteins reactive with patients' autologous sera, of which 14 proteins were identified. Six of the 14 proteins, alpha enolase, pre-B cell-enhancing factor precursor, triosephosphate isomerase, phosphoglycerate mutase 1, fructose-bisphosphate aldolase A, and guanine nucleotide-binding protein beta subunit-like protein, were also up-regulated in hLSCs in the comparative proteomic study, which suggests potential application of these 6 hLSC-associated antigens in diagnosis and therapy of hLSC.     

Comparative proteomics analysis of human lung squamous carcinoma

Two-dimensional polyacrylamide gel electrophoresis (2-DE) profiles of human lung squamous carcinoma tissue and paired surrounding normal bronchial epithelial tissue were compared. Selected differential protein-spots were identified with peptide mass fingerprinting based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and database searching. Well-resolved and reproducible 2-DE patterns of both the tumor and the normal tissues were acquired. The average deviations of spot position were 0.873+/-0.125mm in IEF direction and 1.025+/-0.213mm in SDS-PAGE direction, respectively. For the tumor tissues, a total of 1349+/-67 spots were detected and 1235+/-48 spots were matched with an average matching rate of 91.5%. For the corresponding normal tissues, a total of 1297+/-73 spots were detected and 1183+/-56 spots were matched with an average matching rate of 91.2%. A total of 1069+/-45 spots were matched between the tumor and the normal tissues. Forty differential proteins between tumor and normal tissues were characterized. Some proteins were the products of oncogenes and others were involved in the regulation of cell cycle and signal transduction. These data are valuable for mass identification of differentially expressed proteins involved in lung carcinogenesis, establishing human lung cancer proteome database and screening molecular marker to further study human lung squamous carcinoma.     

Identification of vaccine candidate antigens of Staphylococcus aureus by serological proteome analysis

In this study we applied serological proteome analysis (Klade, C. S. et al. Proteomics 2001, 1, 890-898) for identification of bacterial vaccine candidate antigens. First, approximately one hundred sera from healthy individuals and patients suffering from Staphylococcus aureus infections were screened for antibodies against staphylococcal lysates and recombinant proteins representing surface antigens. Two pools (healthy donors, patients) each consisting of five sera with the highest antiproteinaceous IgG reactivity were selected. Second, S. aureus COL was grown under different conditions and the number of antigens expressed was monitored by Western blot analysis. Third, surface proteins were enriched by digesting the bacterial cell wall under isotonic conditions and subsequent removal of protoplasts. These protein preparations were resolved by two-dimensional electrophoresis (2-DE) (pI 4-7). 2-DE immunoblotting using the preselected serum pools at 1:10 000-1:100 000 dilutions revealed a number of highly immunogenic staphylococcal proteins. Twenty-one spots were isolated by preparative 2-DE, and analysed by matrix-assisted laser desorption/ionization mass spectrometry and tandem mass spectrometry sequencing of tryptic peptides. This led to the identification of 15 proteins including known and novel vaccine candidates. Seroreactivity of several antigens including serine-aspartate repeat containing protein D, immuno-dominant staphylococcal antigen and a novel 309 amino acid lipoprotein was independently confirmed by enzyme-linked immunosorbent assay and Western blot analysis of purified recombinant proteins. In conclusion, serological proteome analysis proved to be a powerful tool for the identification of novel staphylococcal antigens, which provide a basis for rational vaccine design.     

Proteome-Based Analysis of Serologically Defined Tumor-Associated Antigens in Cutaneous Lymphoma

Information on specificities of serological responses against tumor cells in cutaneous lymphoma patients is relatively restricted. To advance the knowledge of serological immune responses against and to assess the scope of tumor antigenicity of cutaneous lymphoma, 1- and 2-dimensional Western blot analyses with sera from patients were combined with proteomics-based protein identification. Testing sera from 87 cutaneous lymphoma patients by 1-dimensional Western blot analysis, 64 cases of seroreactivity against lymphoma cells were found. The positive responses were relatively weak, restricted to few antigens in each case, and heterogeneous. To identify the antigens, proteins of the mycosis fungoides cell line MyLa and primary tumor cells were separated by 2-dimensional gel electrophoresis, Western-blotted and probed with heterogeneous and autologous patient sera. The antigens were identified from silver-stained replica gels by MALDI-TOF mass spectrometry. 14 different antigens were assigned and identified with this proteome-serological approach. Only one, vimentin, had been reported before, the other 13 are new antigens for cutaneous lymphomas.     

Identification of tumor antigens in renal cell carcinoma by serological proteome analysis

We have investigated the suitability of proteomics for identification of tumor-associated antigens. First, we compared the proteomes of nontumorous kidney and renal cell carcinoma (RCC) by two-dimensional gel electrophoresis (2-DE) and silver staining. Protein patterns were markedly different (approximately 800 spots in RCCs versus approximately 1400 spots in kidney). 2-DE immunoblotting revealed five RCC-specific spots, reproducibly reactive with RCC-patient but not healthy donor control sera. Two of these antigens were isolated by preparative 2-DE, and identified by Edman sequencing of tryptic peptides. The first antigen, smooth muscle protein 22-alpha (SM22-alpha), is an actin-binding protein of unknown function predominantly expressed in smooth muscle cells. In situ hybridization revealed that SM22-alpha is not expressed in the malignant cells but in mesenchymal cells of the tumor stroma. The second antigen represents carbonic anhydrase I (CAI), an isoform usually not expressed in kidney. Interestingly, a different isoform (CAXII) has previously been identified by serological expression cloning as an antigen overexpressed in some RCCs. In additional assays, antibodies to recombinant CAI or SM22-alpha were detected in sera from 3/11 or 5/11 RCC patients, respectively, whereas sera from 13 healthy individuals did not react. In conclusion, serological proteome analysis may be a new tool for the identification of tumor-associated antigens.     

Identification of tumor antigens in human lung squamous carcinoma by serological proteome analysis

Autoantibodies against tumor antigens are promising means for cancer diagnosis and prognosis. In this study, we applied a proteomic approach to identify proteins that commonly elicit humoral response in lung squamous carcinoma (LSC). Sera from 20 newly diagnosed patients with LSC and 20 matched healthy individuals were analyzed for antibody-based reactivity against LSC proteins separated by two-dimensional electrophoresis. Autoantibodies against triosephosphate isomerase (Tim) and superoxide dismutase [Mn] (MnSOD) were detected in sera from over 20% patients with LSC but none from the normal controls. Furthermore, the occurrence of autoantibodies against Tim and MnSOD was evaluated by ELISA in an additional 40 LSC patients, 30 other types of cancer (OTC) patients, and 50 noncancer controls (NC). Results showed that frequency of autoantibody against Tim (27.5%) in LSC patients was significantly higher than that in OTC patients (6.7%, p = 0.027) and in NC (6%, p = 0.005). Likewise, frequency of autoantibody against MnSOD in LSC (20%) patients was significantly higher than that in NC (4%, p = 0.016), however, there was no significant difference when comparing to that in OTC patients (6.7%, p = 0.115). We also observed significantly increased expression and secretion of Tim and MnSOD in LSC, which possibly account for their autoantibody development. Our results indicate that autoantibody and antigen of Tim and MnSOD may be useful for screening and diagnosis of the lung squamous carcinoma.     

Optimized serological isolation of lung-cancer-associated antigens from a yeast surface-expressed cDNA library

The technique of serological analysis of antigens by recombinant cDNA expression library (SEREX) uses autologous patient sera as a screening probe to isolate tumor-associated antigens for various tumor types. Isolation of tumor-associated antigens that are specifically reactive with patient sera, but not with normal sera, is important to avoid false-positive and autoimmunogenic antigens for the cancer immunotherapy. Here, we describe a selection methodology to isolate patient sera-specific antigens from a yeast surface-expressed cDNA library constructed from 15 patient lung tissues with non-small cell lung cancer (NSCLC). Several rounds of positive selection using patient sera alone as a screening probe isolated clones exhibiting comparable reactivity with both patient and normal sera. However, the combination of negative selection with allogeneic normal sera to remove antigens reactive with normal sera and subsequent positive selection with patient sera efficiently enriched patient sera-specific antigens. Using the selection methodology described here, we isolated 3 known and 5 unknown proteins, which have not been isolated previously, but and potentially associated with NSCLC.     

Analysis of Leishmania chagasi by 2-D difference gel electrophoresis (2-D DIGE) and immunoproteomic: identification of novel candidate antigens for diagnostic tests and vaccine

Identification of novel antigens is essential for developing new diagnostic tests and vaccines. We used DIGE to compare protein expression in amastigote and promastigote forms of Leishmania chagasi. Nine hundred amastigote and promastigote spots were visualized. Five amastigote-specific, 25 promastigote-specific, and 10 proteins shared by the two parasite stages were identified. Furthermore, 41 proteins were identified in the Western blot employing 2-DE and sera from infected dogs. From these proteins, 3 and 38 were reactive with IgM and total IgG, respectively. The proteins recognized by total IgG presented different patterns in terms of their recognition by IgG1 and/or IgG2 isotypes. All the proteins selected by Western blot were mapped for B-cell epitopes. One hundred and eighty peptides were submitted to SPOT synthesis and immunoassay. A total of 25 peptides were shown of interest for serodiagnosis to visceral leishmaniasis. In addition, all proteins identified in this study were mapped for T cell epitopes by using the NetCTL software, and candidates for vaccine development were selected. Therefore, a large-scale screening of L. chagasi proteome was performed to identify new B and T cell epitopes with potential use for developing diagnostic tests and vaccines.     

Heat-shock protein 27: a potential biomarker for hepatocellular carcinoma identified by serum proteome analysis

Hepatocellular carcinoma (HCC) is the third leading cause of cancer mortality worldwide and ranks second in China. The prognosis of HCC remains dismal mainly because of its late diagnosis, especially in patients with coexisting chronic liver diseases. To identify serum biomarkers for HCC, sera from 20 healthy volunteers, 20 hepatitis B virus (HBV) infected patients and 20 HCC patients were selected for screening study and same number of sera into the same three groups were used for validation study. A strategy including sonication, albumin and immunoglobulin G (IgG) depletion and desalting was optimized for screening differentially expressed proteins of low abundance in serum. By 2-DE image analysis and MALDI-TOF-MS/MS identification, eight proteins including heat-shock protein 27 (HSP27), alpha-fetoprotein (AFP), alpha-1 antitrypsin, clusterin, caeruloplasmin, haptoglobin alpha2 chain, tranferrin and transthyretin were found significantly changed among the healthy, HBV and HCC groups. Further validation study by Western blot showed the detection of HSP27 in 90% HCC sera and two HBV sera, but in none of normal sera. Thus, 2-DE based serum proteome analysis can be useful in the screening of serum biomarkers for HCC and HSP27 could aid in the diagnosis of HCC though further validation is needed.