INPP4B和PTEN在人脑胶质瘤中的表达及意义

目的:检测磷脂酰肌醇4-磷酸酶Ⅱ型（inositol polyphosphate 4-phosphatase type Ⅱ,INPP4B)和磷酸酯酶与张力蛋白同源物（phosphatase and tensin homolog deleted on chromosome ten,PTEN)在人脑胶质瘤中的表达情况及二者的相关性,希望对胶质瘤的诊断分级、预后判断及治疗等方面提供理论依据。方法:采用免疫组织化学法（IHC）和实时荧光定量PCR（qRT-PCR）法分别测定25例正常脑组织和60例不同级别胶质瘤中INPP4B和PTEN蛋白及mRNA的表达情况。结果:在IHC、qRT-PCR中显示:INPP4B、PTEN在正常脑组织组和低级别胶质瘤组、高级别胶质瘤组三组间表达差异均有统计学意义（P＜0.05）;低级别胶质瘤组中 INPP4B、PTEN表达均高于高级别胶质瘤组（P＜0.05）,正常脑组织组中 INPP4B、PTEN表达均高于低级别胶质瘤组和高级别胶质瘤组（P＜0.05）。INPP4B与PTEN mRNA在正常脑组织、低级别胶质瘤、高级别胶质瘤三组中表达呈正相关（P=0.000）。结论:INPP4B与PTEN的表达水平均与胶质瘤的恶性级别呈负相关,随着胶质瘤恶性程度的增高二者的表达水平下降。INPP4B与PTEN在胶质瘤中的表达水平具有正相关性,推测其可能在抑制胶质瘤的发生、发展、侵袭增殖等方面具有协同作用。     

ECT2 mRNA及蛋白在胃癌患者外周血中的表达及意义

目的:探讨ECT2 mRNA及蛋白在胃癌患者外周血中的表达水平及其意义。方法:应用逆转录聚合酶链式反应（RT-PCR）及酶联免疫吸附实验（ELISA）测定50例胃癌患者及30例健康志愿者外周血ECT2 mRNA及蛋白的表达水平。结果:胃癌患者外周血中ECT2 mRNA阳性表达率为54%,健康志愿者外周血中ECT2 mRNA均阴性表达(P<0.05)。ECT2蛋白在胃癌患者外周血中的浓度（1 306.389±215.824)ng/L显著高于健康志愿者外周血中的浓度（502.718±69.440)ng/L(P<0.05),并且与TNM分期、淋巴结转移和浸润深度相关(P<0.05)。结论:外周血中ECT2 mRNA及蛋白的表达与胃癌关系密切,可作为反映胃癌发生、发展过程的有效分子指标。     

CD133 mRNA在胃癌患者外周血单核细胞中的表达及意义

[目的]通过检测胃癌患者外周血单核细胞中CD133的表达,探讨其与临床病理特征的关系。[方法]于术前抽取30例胃癌患者、15例胃溃疡患者、15名健康志愿者外周静脉血,密度梯度离心法分离单核细胞。半定量反转录聚合酶链反应检测CD133 mRNA表达水平。分析胃癌患者外周血CD133 mRNA表达与临床病理特征的关系。[结果]胃癌患者外周血中CD133mRNA表达水平明显高于胃溃疡患者和健康志愿者（0.2804±0.1835 vs 0.0984±0.1321,t=6.724,P〈0.001;0.2804±0.1835 vs 0.0282±0.0597,t=-7.327,P〈0.001）。CD133 mRNA表达与肿瘤最大直径、淋巴结转移、TNM分期有关（P〈0.05）,而与性别、分化程度、淋巴管浸润、血管浸润、浸润深度无关。Spearman相关分析显示,CD133 mRNA表达与淋巴结转移呈正相关（P〈0.01）。[结论]胃癌患者外周血中CD133 mRNA的表达与胃癌的发展、转移及预后密切相关。     

晚期胃癌患者外周血CEA mRNA的表达及临床意义

目的 探讨晚期胃癌患者外周血中CEA mRNA的表达及其临床意义.方法 将临床确诊的32例晚期胃癌根据影像学检查分为血行转移组13例,局部浸润组19例,对照组为胃良性病变10例.以外周血为检测标本,用反转录巢式聚合酶链反应技术(RT-NP-PCR)检测CEA mRNA.结果 初诊时对照组阳件率0%,局部浸润组阳性率63.16%,血行转移组阳性率92.31%,两胃癌组与对照组比较差异有统计学意义(P<0.01),两胃癌组比较差异无统计学意义(P>0.05);6个月后无血行转移病例阳性率50.00%,有血行转移病例阳性率90.00%,两者比较差异有统计学意义(P<0.05).结论 应用RT-NP-PCR技术检测晚期胃癌患者外周血CEA mRNA可作为晚期胃癌血源性播散的指标,对指导选择治疗方案和判断预后有重要意义.     

Survivin mRNA在胃癌外周血中的表达及临床意义

背景与目的:Survivin在肿瘤的发生、发展中起重要作用.近年来,Survivin在胃癌研究领域备受关注.本研究探讨Survivin mRNA在胃癌外周血中的表达及其临床意义.方法:以GAPDH作内参照,应用实时定量RT-PCR技术检测胃炎及化疗前胃癌各50例外周血中Survivin mRNA表达水平.结果:正常对照组(胃炎患者)外周血中未检测到Survivin mRNA表达.胃痛组患者Survivin表达检测率为86%(43例),其中28例出现转移的患者阳性率为100%,22例未出现转移者为68.2%(P＜0.05).Survivin mRNA的表达与年龄、性别及淋巴结转移均无相关性(P＞0.05).回归分析发现,Survivin mRNA的表达水平是影响胃癌预后的独立预测因素.结论:外周血Survivin mRNA可作为临床评估胃癌患者预后的客观指标之一.     

p21活化激酶4在胃癌中的表达及意义

目的探讨p21活化激酶4（PAK4）在人胃癌组织和胃癌细胞系中的表达及与临床病理因素的关系。方法免疫组化法检测95例人胃癌组织中PAK4蛋白的表达；Westernblot法检测40例新鲜胃癌组织和相应的癌旁正常胃黏膜组织以及7种人胃癌细胞系中PAK4蛋白的表达。结果95例人胃癌组织中，PAK4蛋白的阳性表达率为67．4％（64／95），其中62例（96．9％）阳性染色位于细胞浆中，仅有2例（3．1％）位于细胞膜上。PAK4蛋白的阳性表达率与胃癌TNM分期（Χ^2=10．426，P〈0．01）和淋巴结转移（Χ^2=4．912，P〈0．05）有关。新鲜胃癌组织中PAK4蛋白的表达量显著高于癌旁正常胃黏膜组织（t=-5．978，P〈0．01），且随胃癌TNM分期的进展，其表达量逐渐增加。7种人胃癌细胞系中均有PAK4蛋白的表达，与GES-1细胞系相比，PAK4蛋自在ACTS、BGC823、MKN45和N87细胞系中呈高表达，而在MGC803、MKN1和SGC7901细胞系中呈低表达。结论PAK4蛋白的过度表达可能与人胃癌的发生发展及预后有关。     

NF-κB,MMP-7在胃癌中的表达及意义

目的　探讨胃癌中核因子 ΚappaB(NF κB)和MMP 7表达及相互关系。方法　采用免疫组化PV 90 0 0法检测 48例胃癌和 14例正常胃黏膜组织中NF κB亚单位P65蛋白 ,MMP 7蛋白。结果　NF κBP65蛋白和MMP 7蛋白在胃癌中 ,淋巴结转移组 ,低分化胃癌中显著增高 ,且NF κBP65蛋白的表达与MMP 7蛋白的表达密切相关。结论　NF κBP65 ,MMP 7在胃癌的浸润转移和组织分化过程中发挥重要作用 ,NF κB上调MMP 7的表达。     

胃癌中B7-H4和B7-H3的表达及薏苡仁酯对表达的影响

目的:探讨共刺激分子B7-H4和B7-H3在胃癌中的表达,以及薏苡仁酯对这两种分子表达的影响。方法:应用免疫组织化学染色检测胃癌组织中B7-H4蛋白的表达,MTT法检测薏苡仁酯对胃癌细胞株BGC-823的生长抑制效应,RT-PCR检测胃癌组织、胃癌细胞株BGC-823中B7-H4mRNA及B7-H3mRNA的表达,Western Blot检测细胞的B7-H4蛋白及B7-H3蛋白的表达。结果:B7-H4蛋白在80例胃癌组织中有60例阳性表达,阳性率75%。B7-H4蛋白的表达与患者性别、年龄、组织类型、肿瘤大小、淋巴结转移、病理分期、浸润深度无关(P>0.05)。在20例胃癌组织中,B7-H4mRNA表达明显高于癌旁正常组织,B7-H3mRNA表达明显低于癌旁正常组织(P<0.01)。薏苡仁酯可抑制胃癌细胞株BGC-823的生长,呈时间-剂量依赖性,并下调B7-H4 mRNA及蛋白的表达,上调B7-H3mRNA及蛋白的表达(P<0.01)。结论:B7-H4和B7-H3在胃癌异常的表达,可能是肿瘤免疫逃逸的原因之一;薏苡仁酯可影响胃癌细胞B7-H4、B7-H3的表达。     

S100钙结合蛋白A4在胃癌组织中的表达及意义

背景与目的：本研究探讨S100钙结合蛋白A4(S100 calcium-binding protein A4,S100A4)在胃癌旁正常组织、慢性胃炎、肠上皮化生、腺瘤样异型增生和胃癌的组织标本中的表达及与胃癌临床病理特征及预后的关系。方法：利用HE染色对所取胃组织标本进行病理组织学诊断;采用免疫组织化学方法检测标本S100A4蛋白的表达;实时定量聚合酶链反应(quantitative real-time pdymerase chain reaction,qRT-PCR)法检测S100A4 mRNA表达;蛋白[质]印迹法(Western blot)检测S100A4蛋白的表达;采用Kaplan-Meier分析累积生存率。结果：S100A4蛋白和mRNA的表达按以下顺序逐渐增加：胃癌旁正常组织<慢性胃炎<肠上皮化生<腺瘤样异型增生<胃癌。S100A4表达水平与肿瘤大小、浸润深度、淋巴管浸润、淋巴结转移和肿瘤TNM分期有关,但与患者的年龄和性别无关。在较大直径、较深浸润、淋巴结转移的胃癌和肠型癌中可发现S100A4基因的过表达。采用Kaplan-Meier单因素分析显示,弱或中度S100A4基因表达的患者与不表达的患者相比有较低的累积生存率。结论：在胃癌旁正常组织-慢性胃炎-肠上皮化生-腺瘤样异型增生-胃癌过程中,S100A4 mRNA和蛋白表达逐渐增高;S100A4表达与胃癌肿瘤大小、侵袭深度、淋巴结转移和不良预后呈正相关;S100A4过表达参与胃癌发生和演进过程,可以作为反映胃癌恶性程度的生物学指标。     

胃癌中B7-H4和B7-H3的表达及薏苡仁酯对表达的影响

目的：探讨共刺激分子B7-H4和B7-H3在胃癌中的表达,以及薏苡仁酯对这两种分子表达的影响。方法：应用免疫组织化学染色检测胃癌组织中B7-H4蛋白的表达,MTT法检测薏苡仁酯对胃癌细胞株BGC-823的生长抑制效应,RT-PCR检测胃癌组织、胃癌细胞株BGC-823中B7-H4mRNA及B7-H3mRNA的表达,Western Blot检测细胞的B7-H4蛋白及B7-H3蛋白的表达。结果：B7-H4蛋白在80例胃癌组织中有60例阳性表达,阳性率75%。B7-H4蛋白的表达与患者性别、年龄、组织类型、肿瘤大小、淋巴结转移、病理分期、浸润深度无关（P〉0.05）。在20例胃癌组织中,B7-H4mRNA表达明显高于癌旁正常组织,B7-H3mRNA表达明显低于癌旁正常组织（P〈0.01）。薏苡仁酯可抑制胃癌细胞株BGC-823的生长,呈时间-剂量依赖性,并下调B7-H4 mRNA及蛋白的表达,上调B7-H3mRNA及蛋白的表达（P〈0.01）。结论：B7-H4和B7-H3在胃癌异常的表达,可能是肿瘤免疫逃逸的原因之一;薏苡仁酯可影响胃癌细胞B7-H4、B7-H3的表达。     

Basal biomarkers nestin and INPP4B predict gemcitabine benefit in metastatic breast cancer: Samples from the phase III SBG0102 clinical trial

In a formal prospective-retrospective analysis of the phase III SBG0102 clinical trial randomizing metastatic breast cancer patients to gemcitabine-docetaxel or to single agent docetaxel, patients with basal-like tumors by PAM50 gene expression had significantly better overall survival in the gemcitabine arm. By immunohistochemistry (IHC), triple negative status was not predictive, but more specific biomarkers have since become available defining basal-like by nestin positivity or loss of inositol-polyphosphate-4-phosphate (INPP4B). Here, we evaluate their capacity to identify which patients benefit from gemcitabine in the metastatic setting. Nestin and INPP4B staining and interpretation followed published methods. A prespecified statistical plan evaluated the primary hypothesis that patients with basal-like breast cancer, defined as “nestin+ or INPP4B−”, would have superior overall survival on gemcitabine-docetaxel when compared to docetaxel. Interaction tests, Kaplan–Meier curves and forest plots were used to assess prognostic and predictive capacities of biomarkers relative to treatment. Among 239 cases evaluable for our study, 36 (15%) had been classified as basal-like by PAM50. “Nestin+ or INPP4B−” was observed in 41 (17%) of the total cases and was significantly associated with PAM50 basal-like subtype. Within an estimated median follow-up of 13 years, patients assigned as IHC basal “nestin+ or INPP4B-” had significantly better overall survival on gemcitabine-docetaxel versus docetaxel monotherapy (HR = 0.31, 95%CI: 0.16–0.60), whereas no differences were observed for other patients (HR = 0.99), p-interaction < 0.01. In the metastatic setting, women with IHC basal breast cancers defined as “nestin+ or INPP4B-” have superior overall survival when randomized to gemcitabine-containing chemotherapy compared to docetaxel alone. These findings need to be validated using larger prospective-retrospective phase III clinical trials series.     

胃癌患者外周血与胃癌组织中BRCA1基因甲基化的关系及其意义

目的:检测胃癌患者外周血及胃癌组织中BRCA1基因启动子甲基化状态,并探讨两者的关系及意义。方法:采用甲基化特异性PCR(methylation-specific polymerase chain reaction,MSP)技术检测37例胃癌患者外周血及胃癌组织中BRCA1基因启动子甲基化状态。结果:胃癌患者外周血和胃癌组织中BRCA1基因启动子甲基化率分别为40.5%(15/37)和48.6%(18/37),二者相关系数r=0.848(P=0.0004)。结论:胃癌患者外周血与胃癌组织中BRCA1基因启动子存在高比例的甲基化,而且两者甲基化率有良好的一致性。胃癌患者外周血BRCA1基因启动子甲基化的检测具有简便、快捷、可靠的特点,外周血BRCA1基因启动子甲基化有可能成为治疗胃癌的靶点。     

Relationship between LAPTM4B gene polymorphism and susceptibility of gastric cancer.

BACKGROUND: A novel gene called LAPTM4B (lysosome-associated protein transmembrane 4beta) was mapped to 8q22, and contains seven exons. The 2.25-kb messenger RNA of the gene encodes a putative lysosome-associated protein with four transmembrane regions. There are two alleles of the gene, named as LAPTM4B*1 and LAPTM4B*2. Allele *1 differs from allele *2 in that it contains only one copy of a 19-bp sequence in the 5' untranslated region (UTR), whereas this sequence is duplicated and tandemly arranged in allele *2. Studies showed that the allelic variation of LAPTM4B was associated with the genetic susceptibility of hepatocellular carcinoma but not with that of esophageal squamous cell carcinoma. This study was designed to investigate the possible association between the allelic variation of LAPTM4B and the genetic susceptibility of gastric cancer. Materials and methods: The genotype of LAPTM4B was analyzed in 350 unrelated healthy adult individuals and 214 patients with gastric cancer by utilizing polymerase chain reaction based on specific primers. The genotypic distribution of LAPTM4B was analyzed by chi(2) test. RESULTS: The allelic frequencies of the *2 were 33.88% and 24.14% in the gastric cancer group and the healthy control group, respectively, which was significantly different between the two groups (P < 0.001). There was a significant difference in the overall genotypic distribution between the patients and the controls (P = 0.023). The risk of suffering from gastric cancer was increased 1.819 times in the individuals of the *1/2 genotype [95% confidence interval (CI) 1.273-2.601] and 2.387 times in the individuals of the *2/2 genotype of LAPTM4B (95% CI 1.195-4.767) compared with the *1/1 genotype. No association between the genotypic distribution of LAPTM4B and the clinical information on patients of gastric cancer such as age, pathological type, differentiation classification of TNM, and infection of hepatitis B virus was shown. CONCLUSION: This study indicated that allele *2 of LAPTM4B might be the risk factor of gastric cancer, which could be associated with genetic susceptibility of gastric cancer.     

INPP4B is upregulated and functions as an oncogenic driver through SGK3 in a subset of melanomas

Inositol polyphosphate 4-phosphatase type II (INPP4B) negatively regulates PI3K/Akt signalling and has a tumour suppressive role in some types of cancers. However, we have found that it is upregulated in a subset of melanomas. Here we report that INPP4B can function as an oncogenic driver through activation of serum- and glucocorticoid-regulated kinase 3 (SGK3) in melanoma. While INPP4B knockdown inhibited melanoma cell proliferation and retarded melanoma xenograft growth, overexpression of INPP4B enhanced melanoma cell and melanocyte proliferation and triggered anchorage-independent growth of melanocytes. Noticeably, INPP4B-mediated melanoma cell proliferation was not related to activation of Akt, but was mediated by SGK3. Upregulation of INPP4B in melanoma cells was associated with loss of miRNA (miR)-494 and/or miR-599 due to gene copy number reduction. Indeed, overexpression of miR-494 or miR-599 downregulated INPP4B, reduced SGK3 activation, and inhibited melanoma cell proliferation, whereas introduction of anti-miR-494 or anti-miR-599 upregulated INPP4B, enhanced SGK3 activation, and promoted melanoma cell proliferation. Collectively, these results identify upregulation of INPP4B as an oncogenic mechanism through activation of SGK3 in a subset of melanomas, with implications for targeting INPP4B and restoring miR-494 and miR-599 as novel approaches in the treatment of melanomas with high INPP4B expression.     

趋化因子受体CCR5在胃癌患者外周血调节性T细胞上的表达及其临床意义

目的检测胃癌患者外周血中CD4+CD25+CD127-调节性T细胞(Tregs细胞)上的趋化因子受体5(CCR5)的表达水平,并探讨其与胃癌发生、侵袭和转移的相关性。方法利用流式细胞仪检测福建医科大学附属泉州第一医院2013年1月至2014年2月49例胃癌患者与20例健康对照组的外周血Tregs细胞上CCR5的表达水平并进行统计分析。结果胃癌患者组外周血Tregs细胞上CCR5表达水平明显高于健康对照组(P0.05);CCR5在胃癌外周血Tregs细胞上的表达水平与患者年龄、肿瘤直径无关(P0.05),但与组织学分级、病理分期及淋巴结转移相关(P0.05)。多元线性回归分析结果显示:CCR5在胃癌患者外周血Tregs细胞表达水平与性别、年龄、分化程度、肿瘤大小无统计学意义的线性关系,而与病理分期及淋巴结转移呈线性相关。秩相关分析显示:CCR5在胃癌患者外周血Tregs细胞表达水平与性别、年龄、肿瘤大小无统计学意义的相关关系,与病理分期及淋巴结转移呈正线性相关。结论 CCR5在胃癌患者外周血Tregs细胞表达上调,明显高于健康对照组,并与胃癌的病理分期和淋巴结转移关系密切,提示病理分期越高或者淋巴结转移程度越广泛,CCR5在胃癌患者外周血Tregs细胞上表达就越高。CCR5与外周血Tregs细胞的相互介导可能参与机体的肿瘤免疫,在胃癌的发生、发展及侵袭转移中可能起重要作用。