A-kinase interacting protein 1 is sufficiently expressed and positively associates with WHO grade,meanwhile predicts unfavorable overall survival independently in glioma patients

The present study aimed to investigate the association of A-kinase interacting protein 1 (AKIP1) with clinical characteristics, and further explore the prognostic value of AKIP1 in glioma patients.Totally 168 glioma patients who underwent tumor resection were analyzed, and their tumor tissue specimens were acquired for the detection of AKIP1 expression by immunohistochemistry (IHC), which was scored by a semi-quantitative method considering staining intensity and staining density.According to AKIP1 expression in tumor tissues of glioma patients, there were 65 (38.7%) patients with AKIP1 low expression (IHC score 0–3), 48 (28.6%) patients with AKIP1 high + expression (IHC score 4–6), 42 (25.0%) patients with AKIP1 high++ expression (IHC score 7–9) and 13 (7.7%) patients with AKIP1 high+++ expression (IHC score 10–12), respectively. AKIP1 expression was positively associated with World Health Organization grade. Overall survival (OS) was the lowest in the patients with AKIP1 high+++ expression, followed by those with AKIP1 high++ expression and those with AKIP1 high+ expression, and highest in those with AKIP1 low expression. Further subgroup analysis exhibited that AKIP1 expression was negatively associated with OS especially in high-grade glioma patients. In addition, AKIP1 expression was negatively associated with OS in all subgroups of patients with/without adjuvant radiotherapy, with/without adjuvant chemotherapy. Further multivariate Cox''s regression exhibited that AKIP1 high expression was an independent predictive factor for worse OS.AKIP1 presents with the potential to be a novel biomarker for tumor management and prognosis surveillance in glioma patients.     

Transglutaminase 3 expression in hepatocellular carcinoma patients: Correlation with tumor features and survival profile

BackgroundTransglutaminase 3 (TGM3) regulates multiple oncogene pathways (GSK-3β/β-catenin pathway, Akt/ERK pathway, etc.) to promote hepatocellular carcinoma (HCC) cell proliferation, migration and invasion, however, its clinical value for HCC management is still limited. Therefore, we conducted this study to compare the TGM3 expression between tumor tissue and paired adjacent noncancerous tissue, aiming to explore the clinical application of TGM3 in HCC patients.MethodsTotally, 208 HCC patients were enrolled and their clinicopathological features were collected. Then, 208 pairs of HCC specimens and adjacent noncancerous specimens were used to detect TGM3 protein expression by IHC assay and assessed by a semi-quantitative scoring method. Besides, 157 pairs were proposed to detect TGM3 mRNA expression by RT-qPCR.ResultsBoth TGM3 protein (P<0.001) and mRNA (P<0.001) levels were increased in HCC specimens compared to adjacent noncancerous specimens. Besides, TGM3 high protein expression correlated with multifocal tumor nodules (P<0.001), advanced Barcelona Clinic Liver Cancer (BCLC) stage (P = 0.006), higher carcinoembryonic antigen (P = 0.038) and alpha-fetoprotein (AFP) (P<0.001). While TGM3 high mRNA expression correlated with multifocal tumor nodules (P = 0.025), largest tumor size ≥ 5.0 cm (P = 0.042) and higher AFP (P = 0.019). Furthermore, both TGM3 protein (P = 0.002) and mRNA (P = 0.028) high expressions correlated with shorter overall survival (OS). While after adjustment by multivariant Cox's regression, TGM3 protein high expression (vs. low) independently predicted worse OS (P = 0.004).ConclusionsTMG3 expression is increased in tumor tissue, also its high expression correlates with multiple tumor nodules, higher BCLC stage, abnormal AFP and reduced OS in HCC patients.     

CKS2在结直肠癌中的表达和临床意义

背景：细胞周期蛋白依赖性激酶亚基（CKS）家族在细胞周期调节中起重要作用。研究发现其家族成员CKS2在一些恶性肿瘤中呈高表达,并与肿瘤的高侵袭性行为和预后不良相关。目前关于CKS2与结直肠癌关系的文献报道较罕见。目的：研究CKS2在结直肠癌中的表达和临床意义。方法：应用realtimeliT—PCR和蛋白质印迹法检测23例临床结直肠癌手术标本癌组织、癌旁非癌组织和正常组织中的CKS2mRNA和蛋白表达。结果：CKS2mRNA和蛋白在结直肠组织中的相对表达量依次为：癌组织〉癌旁组织〉正常组织,癌组织与正常组织问差异有统计学意义（P〈0．01）。性别对CKS2mRNA在不同结直肠组织中的表达趋势无明显影响。癌组织中的CKS2蛋白表达与肿瘤大小和pTNM分期呈正相关（P〈0．01）,与肿瘤部位无关。结论：CKS2蛋白在结直肠癌中呈高表达并与肿瘤临床病理特征相关,有望成为结直肠癌新的分子标记物和治疗靶点。     

MSI2在胰腺癌中的表达及其临床意义

目的 检测胰腺导管腺癌（PDAC）组织中Musashi 2（MSI2）蛋白和mRNA的表达,分析癌组织MSI2表达与临床病理参数的相关性.方法 应用免疫组化方法检测61例PDAC组织及配对癌旁组织MSI2蛋白表达水平,采用蛋白质印迹法及实时PCR法检测10例癌组织及配对癌旁组织MSI2蛋白及mRNA的表达.分析癌组织MSI2表达与肿瘤临床病理参数的关系.结果 61例PADC患者癌组织MSI2高表达率为63.9％,配对癌旁组织为41.0％,癌组织的表达显著高于配对的癌旁组织,差异有统计学意义（t =2.809,P=0.007）.10例癌组织及癌旁组织的MSI2蛋白表达量分别为0.748±0.195、0.420±0.171;癌组织MSI2 mRNA的表达量为癌旁组织表达量的（2.507±2.981）倍,差异均有统计学意义（t=3.689,P=0.005;t=2.660,P=0.026）.癌组织MSI2表达仅与肿瘤大小呈正相关（x2=5.096,P =0.024）,而与其他参数均无相关性.MSI2高表达患者的术后中位生存期为321 d,低表达者为730 d,高表达者的生存期显著短于低表达者,差异有统计学意义（x2=6.706,P=0.010）.结论 胰腺癌组织中MSI2表达上调,其表达量与肿瘤大小相关,高表达患者的预后差.     

肿瘤相关基因RASAL1在结肠癌中的表达及临床意义

目的:探讨Ras三磷酸鸟苷酶激活样蛋白(RASAL1)及基因在人结肠癌中的表达及其与临床病理参数间的关系.方法:收集2009-04/2010-05经手术切除的结肠腺癌标本50例作为研究组,相应正常组织为对照组,做成蜡块标本;其中20例另取其癌组织、癌旁组织及正常组织的新鲜标本;用免疫组织化学染色观察50例蜡块标本RASAL1蛋白的表达;用RT-PCR检测20例新鲜标本RASAL1 mRNA的表达,并分析其与临床病理参数的关系.结果:RASAL1蛋白主要在细胞质中表达;RASAL1蛋白在结肠癌中的阳性表达率明显低于癌旁组织及正常组织[46%(23/50)vs85%(17/20),96%(48/50),均P<0.05];RASAL1 mRNA在结肠癌中的阳性表达率较癌旁组织及正常组织明显减低[50%(10/20)vs90%(18/20),95%(19/20),均P<0.05],RASAL1蛋白与mRNA的表达呈明显正相关(r=0.686,P<0.01),与肿瘤的分化程度(P<0.05)、侵袭深度(P<0.01)、淋巴结转移(P<0.05)、TNM分期(P<0.05)呈负相关.结论:RASAL1在结肠腺癌组织中低表...     

Expression of Egr-1 gene and its correlation with the oncogene proteins in non-irradiated and irradiated esophageal squamous cell carcinoma

We study the expression of early growth response gene-1 (Egr-1 gene) in non-irradiated and irradiated human esophageal cancer tissues, and its relationship with the expression of C-fos, C-jun onco-proteins as well as Egr-1 target gene proteins P53, Rb and Bax expression. In situ hybridization (ISH) and immunohistochemistry (IHC) were used respectively to detect Egr-1 mRNA, Egr-1, C-fos, C-jun, P53, Rb and Bax proteins in 80 surgically resected non-irradiated and irradiated tumor specimens of esophageal squamous cell carcinoma. Egr-1 gene mRNA and Bax protein were located in the cytoplasm, whereas Egr-1, C-fos, C-jun, P53, Rb proteins were located in the nuclei. Egr-1 was expressed in nine out of 40 cases (22.5%) of non-irradiated and 23 of 40 cases (57.5%) of irradiated tumor specimens. No correlation was found between Egr-1 gene expression and C-fos, C-jun onco-proteins expression, neither was any correlation disclosed between Egr-1 gene expression with its target gene protein expression. Patients who underwent radiotherapy with Egr-1 overexpressed in their cancer tissue had better prognosis. Radiotherapy up-regulates Egr-1 expression in esophageal carcinoma. Egr-1 overexpression may be a potential radiation response gene marker and may play an important role in prognosis of esophageal squamous cell carcinoma.     

Musashi 2 (MSI2) expression as an independent prognostic biomarker in non-small cell lung cancer (NSCLC)

BackgroundMusashi-2 (MSI2) is a member of RNA-binding protein family that regulates mRNA translation of numerous intracellular targets and influences maintenance of stem cell identity. This study assessed MSI2 as a potential clinical biomarker in non-small cell lung cancer (NSCLC).MethodsThe current study included 40 patients with NSCLC, of whom one presented with stage 1, 14 presented with stage II, 15 presented with stage III, and 10 patients had stage IV. All patients received standard of care treatments. All patient samples were obtained before treatment started. We used immunohistochemical (IHC) approach to measure MSI2 protein expression in matching specimens of normal lung versus tumor tissues, and primary versus metastatic tumors, followed by correlative analysis in relation to clinical outcomes. In parallel, clinical correlative analysis of MSI2 mRNA expression was performed in silico using publicly available datasets (TCGA/ICGC and KM plots).ResultsMSI2 protein expression in patient samples was significantly elevated in NSCLC primary tumors versus normal lung tissue (P=0.03). MSI2 elevated expression positively correlated with a decreased progression free survival (PFS) (P=0.026) combined for all stages and with overall survival (OS) at stage IV (P=0.013). Elevated MSI2 expression on RNA level was confirmed in primary tumor versus normal tissue samples in TCGA dataset (P<0.0001), and positively correlated with decreased OS (P=0.02). No correlation was observed between MSI2 expression and age, sex, smoking, and treatment type.ConclusionsElevated MSI2 expression in primary NSCLC tumors is associated with poor prognosis and can be used as a novel potential prognostic biomarker in NSCLC patients. Future studies in an extended patient cohort are warranted.     

15-脂氧合酶-1在胃癌中的表达及其临床意义

目的检测15-脂氧合酶-1（15-LOX-1）mRNA及蛋白在胃癌及癌旁正常组织中的表达，并探讨15-LOX-1表达与胃癌临床病理因素的关系。方法采用RT-PCR、westernblot和免疫组织化学方法检测胃癌组织及相匹配的癌旁正常组织中15-LOX-1 mRNA和蛋白的表达。结果15-LOX-1 mRNA及蛋白在胃癌组织中的表达均显著低于癌旁正常组织（P＜0．05）。15-LOX-1蛋白表达水平与胃癌中肿瘤大小、淋巴结转移和TNM分期呈明显的负相关（P＜0．05）。结论15-LOX-1蛋白可能对胃癌的发生发展有一定抑制作用。检测胃癌中15-LOX-1的表达情况对评估患者的预后可能具有意义。     

The relationship between SPARC expression in primary tumor and metastatic lymph node of resected pancreatic cancer patients and patients' survival

BACKGROUND:Previous researches in pancreatic cancer demonstrated a negative correlation between secreted protein acidic and rich in cysteine (SPARC) expression in primary tumor and survival,but not for SPARC expression in lymph node.In the present study,we aimed to evaluate the SPARC expression in various types of tissues and its impact on patients' prognosis.METHODS:The expression of SPARC was examined by immunohistochemistry in resected pancreatic cancer specimens.Kaplan-Meier analyses and Cox proportional hazards regression were applied to assess the mortality risk.RESULTS:A total of 222 tissue samples from 73 patients were collected to evaluate the SPARC expression,which included 73 paired primary tumor and adjacent normal tissues,38 paired metastatic and normal lymph nodes.The proportion of positive SPARC expression in metastatic lymph node was high (32/38),whereas in normal lymph node it was negative (0/38).Positive SPARC expression in primary tumor cells was associated with a significantly decreased overall survival (P=0.007) and disease-free survival (P=0.003),whereas in other types of tissues it did not show a predictive role for prognosis.Univariate and multivariate analyses both confirmed this significance.CONCLUSION:SPARC can serve a dual function role as both predictor for prognosis and potentially biomarker for lymph node metastasis in resected pancreatic cancer patients.     

基于TCGA和GEO数据挖掘分析NAT1在结肠癌中的表达及预后意义

目的探讨结肠癌癌组织中N-乙酰化转移酶1（NAT1）的表达及其对结肠癌患者预后的影响。 方法通过肿瘤免疫评估资源（TIMER）数据库分析NAT1 mRNA在33种肿瘤中的表达情况，并用人类蛋白质图谱（HPA）数据库的免疫组化结果验证NAT1蛋白在结肠癌中的表达。通过肿瘤基因图谱（TCGA）和基因表达综合（GEO）数据库获得NAT1在结肠癌中的表达数据及相关临床特征参数，分析NAT1 mRNA表达水平与结肠癌患者的临床特征和总生存期（OS）的关系，并构建预后模型。采用基因集富集分析（GSEA）预测NAT1相关的基因通路。采用CIBERSORT分析NAT1与免疫浸润的关系。 结果TIMER数据库分析结果显示，在13种肿瘤组织中NAT1 mRNA表达水平低于正常对照组织。TCGA数据库结果提示，结肠癌组织中NAT1 mRNA表达水平均明显低于正常对照组织或癌旁正常组织，差异均有统计学意义（均P＜0.01），并在GSE44076、GSE44861和GSE73360中得到验证。HPA数据库的免疫组化结果提示，NAT1蛋白在结肠癌组织中呈低表达。TCGA数据库分析结果提示，NAT1 mRNA表达水平与结肠癌患者的N分期、M分期和stage分期均有关（均P＜0.01）。NAT1高表达组患者OS均好于低表达组（均P＜0.05）。单因素Cox分析表明，NAT1 mRNA表达水平是影响结肠癌患者OS的危险因素（P＜0.05），并和其他危险因素构建列线图，同时使用校准曲线和ROC评估了预后模型的特异性和敏感性。选取本院确诊的35例结肠癌患者肿瘤组织作为肿瘤组，选取其癌旁正常组织作为对照组。采用实时荧光定量PCR（qRT-PCR）法检测NAT1表达水平，结果与数据库结果一致（P＜0.05）。GSEA结果提示NAT1高表达样本上调抗坏血酸和醛糖酸盐代谢、戊糖和葡萄糖醛酸的相互转化、淀粉和蔗糖代谢、卟啉和叶绿素代谢途径、视黄醇代谢、药物代谢相关酶等基因集，而下调糖胺聚糖生物合成途径、Hedgehog信号通路、基底细胞癌、ECM受体作用通路、神经活性配体-受体相互作用途径、Wnt信号通路等基因集。使用CIBERSORT计算每个样品中的免疫细胞浸润，高NAT1组免疫细胞中原始B细胞、静息记忆CD4 T细胞、静息树突状细胞、活化树突状细胞显著增加，而M0巨噬细胞显著减少（均P＜0.05）。 结论结肠癌中NAT1 mRNA表达水平下调，NAT1低表达提示结肠癌患者的预后差，可作为结肠癌患者治疗的潜在靶点。     

Decreased expression of insulin-like growth factor binding protein 7 in human colorectal carcinoma is related to DNA methylation

Purpose Insulin-like growth factor binding protein 7 (IGFBP-7) is considered a tumor suppressor in various cancers, but its role in colorectal cancer (CRC) is still uncertain. The aims of this study were to analyze the IGFBP-7 expression, and explore the mechanism responsible for the inactivation of IGFBP-7 in CRC. Methods mRNA expression was studied by RT-PCR and Northern blot analysis of cultured cells. Methylation status was analyzed by treatment with 5-aza-2′-deoxycytidine followed by sequencing of PCR products of sodium bisulfite-treated genomic DNA. IGFBP-7 protein expression was evaluated by immunohistochemistry (IHC) on tissue microarrays. Results mRNA expression was lost in six out of eight CRC cell lines as compared to normal colon cells. DNA methylation was found in the region of exon 1 and intron 1 of IGFBP-7. In tumor tissue, 107 out of 279 samples showed a negative expression of IGFBP-7 by IHC, which was significantly associated with poor prognosis. The analysis of 37 paired cancerous and normal mucosa samples confirmed the downregulation in the tumors, but revealed variable basal expression levels of IGFBP-7 in normal mucosal samples. Conclusions DNA methylation is a mechanism responsible for IGFBP-7 gene silencing providing a target for therapeutic intervention of this tumor suppressor gene.     

Angiopoietins and Tie-2 expression in angiogenesis and proliferation of human hepatocellular carcinoma

Human hepatocellular carcinoma (HCC) is a hypervascular tumor but the mechanisms underlying the process of angiogenesis are not fully understood. Angiopoietins (Ang) have been recently identified as ligands for Tie-2 receptor and are thought to be important factors in vascular maturation and stability during angiogenesis. In this study, we investigated the expression of Ang-1, Ang-2, Tie-2, and vascular endothelial growth factor (VEGF) in surgically resected specimens from 46 patients with HCC to determine their potential role in tumor angiogenesis and its progression. VEGF messenger RNA (mRNA) was significantly up-regulated in HCC compared to normal liver tissue from patients with hepatic metastases. No differences were found between HCC and adjacent liver tissue. Meanwhile, Ang-2 mRNA expression in HCC was significantly increased when compared to adjacent liver tissue. On the other hand, Ang-1 and Tie-2 mRNA expression in HCC was not different from that in adjacent liver tissue. Immunohistochemical staining also showed increased Ang-2 protein in HCC. Furthermore, a high Ang-2/1 mRNA ratio in HCC was closely associated with tumor portal vein invasion, tumor diameter, and the microvessel density level as assessed by CD34 immunostaining. With regard to prognosis, the survival time for patients in the high Ang-2/1 mRNA ratio group was significantly poorer when compared with the low Ang-2/1 mRNA ratio group. In conclusion, an increased expression of Ang-2/1 in the presence of VEGF may play a critical role in promoting tumor angiogenesis and progression in human HCC.     

Hypermethylation and aberrant expression of Wnt antagonist secreted frizzled-related protein 1 in gastric cancer

AIM： To identify the methylation of secreted frizzled-related protein 1 （SFRP1） in gastric cancer and to investigate the aberrant expression of SFRP1 and its correlation with the clinical pathological features of patients.
METHODS： We determined SFRP1 methylation and SFRP1 mRNA expression in 3 gastric cancer cell lines SGC-7901, BGC-823, HGC-27, from 52 primary gastric cancer specimens and matched tumor adjacent tissue specimens by methylation-specific （MSP） PCR and RT-PCR respectively. Fisher＇s exact test was used to analyze the statistical association between clinical pathological data and aberrant expression of SFRP1.
RESULTS： In 3 cancer cell lines, BGC-823 and HGC-27 had methylated SFRP1 and lost SFRP1 mRNA expression. After treatment of BGC-823 and HGC-27 with the demethylating agent, 5-aza-2′-deoxycytidine, SFRP1 was re-expressed. In 52 primary gastric cancer specimens and matched tumor adjacent tissue specimens, hypermethylation of SFRP1 was detected in 23 （44%） and 8 （15%） specimens respectively （x^2= 10.34, P 〈 0.01）. Loss of SFRP1 expression was detected in 17（33%） and 6 （12%） specimens respectively （x^2= 6.75, P 〈 0.01）. There was a significant correlation between SFRP1 hypermethylation and SFRP1 expression loss. SFRP1 expression was also correlated significantly with tumor stage and lymph node status, but not with patient sex, age and histological type.
CONCLUSION： SFRP1 inactivation is a common and early event caused mainly by hypermethylation in gastric cancer. SFRP1 expression loss may be correlated with tumor metastasis in primary gastric cancer.     

Prognostic significance of HIF-2α/EPAS1 expression in hepatocellular carcinoma

AIM: To evaluate the prognostic significance of HIF-2α/EPAS1 expression in hepatocellular carcinoma(NCC).METHODS: Surgical specimens from 315 patients with HCC as well as 196 adjacent noncancerous lesions and 22 cases of normal liver tissue were investigated by immunohistochemistry (IHC) for HIF-2α/EPAS1 using a standard detection system. Correlations with clinicopathological factors, VEGF, microvessel density(MVD), and prognosis were analyzed.RESULTS: Immunoreactivity of HIF-2α/EPAS1 was positive in 69.5% of HCC, 55.6% of adjacent noncancerous tissue, and 0% of normal liver tissue. And it was significantly correlated with tumor grade, venous invasion, intrahepatic metastasis, necrosis, and capsule infiltration. Correlation analysis of HIF-2α/EPAS1 with angiogenic factor VEGF (P＜0.001), and MVD (P =0.016) was also noted. HIF-2α/EPAS1 protein was less frequently expressed in low MVD cases, whereas a high rate of expression was noted in cases with both medium and high MVD (P=0.042). By Kaplan-Meier analysis,strong HIF-2α/EPAS1 staining (＞ 50% of tumor cells) in HCC correlated with a shortened survival in patients (Cox's regression, P＜0.001, r= 3.699).CONCLUSION: We conclude that HIF-2α/EPAS1 expression may play an important role in tumor progression and prognosis of HCC. Assessment of HIF-2α/EPAS1 expression in HCC may be used as a diagnostic tool and possibly a target in the treatment of HCC.     

Prognostic signif icance of HIF-2α/EPAS1 expression in hepatocellular carcinoma

AIM: To evaluate the prognostic signif icance of HIF- 2α/EPAS1 expression in hepatocellular carcinoma (HCC). METHODS: Surgical specimens from 315 patients with HCC as well as 196 adjacent noncancerous lesions and 22 cases of normal liver tissue were investigated by immunohistochemistry (IHC) for HIF-2α/EPAS1 using a standard detection system. Correlations with clinicopathological factors, VEGF, microvessel density (MVD), and prognosis were analyzed. RESULTS: Immunoreactivity of HIF-2α/EPAS1 was positive in 69.5% of HCC, 55.6% of adjacent noncancerous tissue, and 0% of normal liver tissue. And it was significantly correlated with tumor grade, venous invasion, intrahepatic metastasis, necrosis, and capsule infiltration. Correlation analysis of HIF-2α/EPAS1 with angiogenic factor VEGF (P < 0.001), and MVD (P = 0.016) was also noted. HIF-2α/EPAS1 protein was less frequently expressed in low MVD cases, whereas a high rate of expression was noted in cases with both medium and high MVD (P = 0.042). By Kaplan-Meier analysis, strong HIF-2α/EPAS1 staining (> 50% of tumor cells) in HCC correlated with a shortened survival in patients (Cox's regression, P < 0.001, r = 3.699). CONCLUSION: We conclude that HIF-2α/EPAS1 expression may play an important role in tumor progression and prognosis of HCC. Assessment of HIF-2α/EPAS1 expression in HCC may be used as a diagnostic tool and possibly a target in the treatment of HCC.