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。     

15-脂氧合酶在肿瘤中的作用

15-脂氧合酶(15-LOX)是催化不饱和脂肪酸代谢的关键酶之一.人类中存有两型15-LOX:15-LOX-1和15-LOX-2,两者在催化性质、组织分布及肿瘤生物学功能上有所差异.近年来大量研究发现15-LOX在结肠癌、前列腺癌、食道癌等恶性肿瘤表达较正常组织降低,能够抑制肿瘤生长,并使其发生凋亡,在肿瘤发生发展中起着重要作用.以15-LOX为靶向的肿瘤防治及其临床应用具有广阔的前景.     

15-脂氧合酶-1 mRNA、Podoplanin在食管鳞癌中的表达及临床意义

目的 探讨15-脂氧合酶-1(15-LOX-1)和Podoplanin在食管鳞癌(ESCC)中的表达及其与肿瘤淋巴结转移的关系.方法 RT-PCR法测定45例ESCC患者癌组织和相邻癌旁正常组织中15-LOX-1 mRNA的表达,免疫组化法检测Podoplanin的表达,计数淋巴管密度(LVD).结果 15-LOX-1 mRNA在45例ESCC的表达率为28.9%(13/45),明显低于相应癌旁正常组织73.3%(33/45),P<0.05.Podoplanin特异表达于淋巴管内皮,多位于癌巢周围间质中.40例ESCC中,淋巴结转移组LVD均值高于无淋巴结转移组(P=0.019).15-LOX-1 mRNA和LVD均值与肿瘤分化程度、淋巴结转移有关.结论 15-LOX-1表达下调与食管鳞癌分化程度和病期进展有关.肿瘤内部存在的新生淋巴管可能参与了肿瘤淋巴道转移.Podoplanin是较特异的淋巴内皮标志物,通过检测Podoplanin的表达,评价淋巴管生成,可能成为预测患者淋巴结转移和病情进展的手段之一.     

胃癌组织中核因子-κB和环氧合酶-2的表达及其临床意义

背景:有关核因子(NF)-κB和环氧合酶(COX)-2在肿瘤组织中表达的研究虽然较多,但其临床意义和相互关系尚无定论.目的:研究NF-κB和COX-2在胃癌组织中的表达及其关系.方法:采用免疫组化SP法检测142例胃癌组织中NF-κB和COX-2蛋白的表达,以相应的癌旁正常组织(30例)作对照.结果:NF-κB在胃癌组织中的阳性表达率为62.0%,显著高于对照组(P＜0.01);NF-κB的表达与组织学类型、淋巴结转移和远处转移的临床指标呈显著相关(P＜0.05).COX-2在胃癌组织中的阳性率为64.1%,在对照组中无表达,两者比较有显著性差异(P＜0.01),且在淋巴结转移组阳性率显著高于无转移组(P＜0.01).胃癌组织中NF-κB和COX-2的表达呈显著正相关(r=0.380,P＜0.01).结论:NF-κB和COX-2在胃癌的发生、发展中起重要作用,NF-κB可能促进了COX-2的表达.     

12-脂氧合酶在胰腺癌中的表达及其临床意义

目的 检测胰腺癌组织和胰腺癌细胞株SW1990和PANC1的12-脂氧合酶(12-lipooxygenase,12-LOX)表达,探讨其与胰腺癌临床病理参数的关系.方法 分别应用免疫组化染色、RT-PCR和蛋白质印迹法检测胰腺癌组织、癌旁正常胰腺组织及胰腺癌细胞株SW1990和PANC1中12-LOX mRNA和蛋白的表达.分析胰腺癌组织12-LOX表达与临床病理参数的相关性.结果 30例胰腺癌组织12-LOX表达阴性8例,弱阳性7例,强阳性15例,总阳性率为73.3％.SW1990和PANC1细胞均表达12-LOX.阳性表达的胰腺癌组织及两株胰腺癌细胞株均表达12-LOX mRNA,而癌旁正常胰腺组织不表达12-LOX mRNA及蛋白.胰腺癌组织12-LOX强阳性表达与肿瘤TNM分期、肿瘤病理分级和淋巴结转移相关(P＜0.05),而与患者年龄、性别无关.结论 12-LOX在胰腺癌中表达上调,与胰腺癌的恶性生物学行为有关.     

15-脂氧合酶-1对胃癌细胞周期的影响及其机制研究

目的研究15-脂氧合酶-1(15-LOX-1)对胃癌细胞AGS细胞周期的影响,并探讨其作用机制。方法实验分为15-LOX-1重组质粒转染组、空载体组和AGS组,转染胃癌细胞AGS,用RT-PCR和Western blot检测15-LOX-1 mRNA和蛋白的表达,流式细胞术检测AGS细胞周期的分布,Western blot检测S相激酶相关蛋白2(S-phase Kinase associated protein 2,Skp2)和细胞周期抑制因子P27的表达。结果重组质粒转染组AGS胃癌细胞分别从mRNA和蛋白水平检测到15-LOX-1的表达,流式细胞仪结果显示15-LOX-1可使肿瘤细胞大部分滞留于G0/G1期,进入S期的细胞减少,S期比例下降,并且转染15-LOX-1后P27表达上调,而Skp2表达下降(P0.05)。结论 15-LOX-1可能通过调节细胞周期蛋白Skp2和P27的表达,使细胞周期阻滞于G0/G1期,抑制肿瘤细胞增殖。     

血红素氧合酶-1在胃癌腹膜转移中的表达及意义

目的 探讨血红素氧合酶-1(hemeoxygenase-1,HO-1)在胃腺癌及腹膜转移灶、细胞系及耐药细胞系中的表达及意义.方法 用免疫组化法检测68例胄腺癌组织及其腹膜转移灶、转移灶旁无瘤腹膜组织中HO-1的表达,以及46例无腹膜转移的胃腺癌组织中HO-1的表达.Western印迹法检测胃腺癌腹膜转移组织及耐药细胞系HO-1的表达.结果 胃腺癌及其腹膜转移灶HO-1的阳性表达率分别为39.7%(27/68)和41.2%(28/68),显著高于癌旁无瘤腹膜组织[0%(0/68),P<0.01],亦显著高于无腹膜转移胃癌组织[21.7%(10/46),P<0.05].低分化转移灶HO-1表达水平显著高于中、高分化转移灶(P<0.05).Western印迹法检测胃癌腹膜转移患者的转移灶HO-1表达水平显著高于其癌旁无瘤腹膜组织(P<0.05).HO-1在耐药细胞系GC9811-P的表达水平较其亲本细胞系明显上调(P<0.05).结论 HO-1在胃腺癌腹膜转移过程中表达增高,HO-1可能参与胃癌腹膜转移发生.高表达HO-1与胃癌腹膜转移组织的恶性程度有关,其信号转导通路可能存在于组织的上皮细胞,并可能与多药耐药有关.     

血红素氧合酶-1在胃癌腹膜转移中的表达及意义

目的 探讨血红素氧合酶-1(hemeoxygenase-1,HO-1)在胃腺癌及腹膜转移灶、细胞系及耐药细胞系中的表达及意义.方法 用免疫组化法检测68例胄腺癌组织及其腹膜转移灶、转移灶旁无瘤腹膜组织中HO-1的表达,以及46例无腹膜转移的胃腺癌组织中HO-1的表达.Western印迹法检测胃腺癌腹膜转移组织及耐药细胞系HO-1的表达.结果 胃腺癌及其腹膜转移灶HO-1的阳性表达率分别为39.7%(27/68)和41.2%(28/68),显著高于癌旁无瘤腹膜组织[0%(0/68),P<0.01],亦显著高于无腹膜转移胃癌组织[21.7%(10/46),P<0.05].低分化转移灶HO-1表达水平显著高于中、高分化转移灶(P<0.05).Western印迹法检测胃癌腹膜转移患者的转移灶HO-1表达水平显著高于其癌旁无瘤腹膜组织(P<0.05).HO-1在耐药细胞系GC9811-P的表达水平较其亲本细胞系明显上调(P<0.05).结论 HO-1在胃腺癌腹膜转移过程中表达增高,HO-1可能参与胃癌腹膜转移发生.高表达HO-1与胃癌腹膜转移组织的恶性程度有关,其信号转导通路可能存在于组织的上皮细胞,并可能与多药耐药有关.     

15-LOX-1基因表达对胃癌细胞增殖的抑制作用

目的研究15-LOX-1基因对人胃癌细胞增殖的影响。方法通过脂质体介导瞬时转染15-LOX-1基因于培养的胃癌细胞株（AGS）中，以转染空载体pcDNA3．1（＋）的细胞及未转染质粒的细胞作为对照；用RT—PCR和Westernblot检测人胃癌细胞的15-LOX-1mRNA及蛋白表达；倒置显微镜观察转染前后AGS形态变化；用MTT法比较转染前后AGS的增殖水平，流式细胞术检测转染后AGS细胞周期的变化。结果胃癌细胞系巾未检测到15-LOX—1mRNA和蛋白的表达，转染后的AGS表达有15-LOX—1的mRNA及蛋白。转染重组质粒绀细胞增殖显著低于未转染组及空质粒转染组（P〈0．05），转染后AGS细胞G0／G1期细胞明显增加，S期细胞明显减少，与未转染组及空质粒转染组相比均有显著性差异（P〈0．05）。结论15-LOX-1基因能抑制胃癌细胞的增殖，为进一步探讨胃痛的发病机制及治疗提供实验依据。     

Regulation of Akt/PKB activity by P21-activated kinase in cardiomyocytes

Akt/PKB is a critical regulator of cardiac function and morphology, and its activity is governed by dual phosphorylation at active loop (Thr308) by phosphoinositide-dependent protein kinase-1 (PDK1) and at carboxyl-terminal hydrophobic motif (Ser473) by a putative PDK2. P21-activated kinase-1 (Pak1) is a serine/threonine protein kinase implicated in the regulation of cardiac hypertrophy and contractility and was shown previously to activate Akt through an undefined mechanism. Here we report Pak1 as a potential PDK2 that is essential for Akt activity in cardiomyocytes. Both Pak1 and Akt can be activated by multiple hypertrophic stimuli or growth factors in a phosphatidylinositol-3-kinase (PI3K)-dependent manner. Pak1 overexpression induces Akt phosphorylation at both Ser473 and Thr308 in cardiomyocytes. Conversely, silencing or inactivating Pak1 gene diminishes Akt phosphorylation in vitro and in vivo. Purified Pak1 can directly phosphorylate Akt only at Ser473, suggesting that Pak1 may be a relevant PDK2 responsible for AKT Ser473 phosphorylation in cardiomyocytes. In addition, Pak1 protects cardiomyocytes from cell death, which is blocked by Akt inhibition. Our results connect two important regulators of cellular physiological functions and provide a potential mechanism for Pak1 signaling in cardiomyocytes.     

Akt和磷酸化Akt1在胰腺癌组织中的表达及意义

目的 了解Akt和磷酸化Akt1(p-Akt1)蛋白在胰腺癌组织中的表达情况,探讨其临床意义.方法 应用免疫组织化学法检测74例胰腺癌组织、10例正常胰腺组织中Akt和p-Akt1蛋白的表达,并分析其与临床病理学特征及预后的关系.结果 Akt和p-Akt1蛋白在胰腺癌组织中的阳性表达率分别为87.8%和83.8%,而正常胰腺组织均阴性表达,相差非常显著(P<0.05).癌组织中Akt和p-Akt1蛋白的表达呈正相关性(r=0.274,P=0.018).Akt和p-Akt1蛋白表达与年龄、性别、肿瘤生长部位、肿瘤大小、病理学分级、淋巴结转移、临床分期及神经浸润均无显著相关性(P>0.05),但p-Akt1蛋白高表达与病理学T分期及TNM分期相关(P=0.002).Akt和p-Akt1蛋白高表达者的中位生存期分别为(16.0±5.7)个月和(23.0±5.5)个月,明显高于低表达者的(9.3±0.2)个月和(11.1±1.8)个月(P分别为0.007和0.004).结论 胰腺癌组织中p-Akt1蛋白表达的程度与良性预后有关,检测胰腺癌中p-Akt1蛋白的表达具有一定的临床意义.     

PKBα/Akt1基因多态性与2型糖尿病的相关性研究

目的 研究蛋白激酶B α亚型(PKBα,也称Akt1)基因单核苷酸多态性(SNP)与上海地区汉族人群2型糖尿病易感性的关系.方法 利用等位基因特异PCR技术对460例2型糖尿病患者及444名正常对照者(NC组)Akt1基因3个标签SNP位点rs2494743、rs2494738和rs3001371进行基因分型.结果 位点rs2494738和rs3001371的基因型分布在糖尿病组和NC组之间呈现显著性差异(均P＜0.01);rs2494738和rs3001371等位基因频率在2型糖尿病组和NC组的分布也呈现显著性差异(均P＜0.01);rs2494738的多态性与2型糖尿病发生风险呈等位基因计量效应关系.但rs2494743基因型和等位基因分布在NC组与2型糖尿病组中差异无统计学意义.单倍型分析结果显示3个单倍型频率在2型糖尿病组和NC组之间也存在显著差异(均P＜0.01).结论 在上海地区的汉族人群中,Akt1基因可能是2型糖尿病的易感基因之一;其SNP位点rs2494738和rs3001371变异可能与2型糖尿病发病相关.     

The 15-lipoxygenase-1 expression may enhance the sensitivity to non-steroidal anti-inflammatory drug-induced apoptosis in colorectal cancers from patients who are treated with the compounds

BACKGROUND AND AIM: Non-steroidal anti-inflammatory drugs (NSAIDs) can prevent colorectal cancer (CRC), but their effect is limited. Recent studies have shown the involvement of 15-lipoxygenase-1 (15-LOX-1) in NSAID-induced apoptosis in colorectal carcinoma cells. We evaluate whether 15-LOX-1 expression influences the sensitivity of NSAID-induced apoptosis in CRCs. METHODS: In 22 CRC surgical samples from NSAID users who had been constant for more than 5 years and 28 CRC surgical samples from NSAID non-users, the expressions of 15-LOX-1, cyclooxygenase-2 (COX-2), beta-catenin, and p53 were analyzed using immunohistochemistry. TUNEL assay was also performed for samples. The effects of the transient transfection of 15-LOX-1 cDNA on indomethacin-induced apoptosis were certified in HCT-116 cells. The effects of adding 13-S-hydroxyoctadecadinoic acid (13-S-HODE) on indomethacin-induced apoptosis were also examined in HCT-116 cells. The levels of apoptosis were determined by the analysis of the floating-cells ratio and DNA gel electrophoresis. RESULTS: The expression of 15-LOX-1 on CRCs from NSAID users was significantly decreased compared with those from NSAID non-users; however, the expressions of other molecules were not significantly different between two groups. The levels of TUNEL scoring in samples from NSAID users were similar to those from NSAID non-users. Indomethacin (100 microM) induced less apoptosis in mocked cells, whereas the same concentrations of indomethacin enhanced the level of apoptosis in 15-LOX-1-transfected cells. 13-S-HODE also increased the level of indomethacin-induced apoptosis in cells. CONCLUSION: Results suggest that 15-LOX-1 expression may be one of the mechanisms which enhance the sensitivity to NSAID-induced apoptosis in CRCs from patients who are treated with the compounds.     

Bmi-1基因对胃癌细胞增殖的影响及机制

目的:探索干扰Bmi-1基因后对其可能的下游基因Akt/PKB活性和P16INK4a基因表达的影响及对肿瘤细胞增殖和细胞衰老的作用.方法:用s iRNA技术干扰Bmi-1表达后,运用Western blot检测Bmi-1蛋白及相关蛋白pAkt、Akt和P16INK4a的表达,同时进行SA-β-Gal染色检测细胞衰老,软琼脂克隆形成实验检测细胞的增殖能力.结果:转染Bmi-1 i质粒组平均细胞衰老率28%±3.5%,而对照Ctrl i组为16%±2.7%,有明显统计学差异( P<0.01).转染Bmi-1 i质粒组细胞平均克隆形成数为3.4±1.4个,而对照Ctrl i组为11±2.3个,两组比较有明显的统计学差异( P<0.01).Bmi-1 i 组较Ctrl i 组Bmi-1和pAkt蛋白表达明显下降,而P16INK4a蛋白表达升高.结论:干扰Bmi-1可以通过降低Akt/PKB活性和上调P16INK4a蛋白表达,促进肿瘤细胞衰老并减弱肿瘤细胞的增殖能力.     

Expression and clinical significance of CD73 and hypoxia-inducible factor-1α in gastric carcinoma

AIM: To investigate the expression of CD73 and hypoxia-inducible factor-1α (HIF-1α) in human gastric carcinoma, and explore their clinical significance and prognostic value. METHODS: CD73 and HIF-1α expressions were detected by immunohistochemistry in consecutive sections of tissue samples from 68 gastric carcinoma patients. The peritumor tissues 2 cm away from the tumor were obtained and served as controls. The presence of CD73 and HIF-1α was analyzed by immunohis-tochemistry using the Envision technique. RESULTS: CD73 and HIF-1α expressions in gastric carcinoma were significantly higher than those in gastric mucosal tissues as control (P < 0.001) and showed a close correlation (Spearman r = 0.390, P = 0.001). Overexpression of CD73 was positively correlated with differentiation of tumor (P = 0.000), histopathology (P = 0.041), depth of invasion (P < 0.001), nodal status (P = 0.003), metastasis (P = 0.013), and the American Joint Committee on Cancer (AJCC) stage (P < 0.001). High expression of HIF-1α was positively correlated with tumor diameter (P = 0.031), depth of invasion (P = 0.022), and AJCC stage (P = 0.035). The overall survival rate was low in the patients with high expression of CD73 (P < 0.001). Moreover, CD73+/HIF-1α+ patients had the worst prognosis (P < 0.001). CD73 expression was proven to be an independent predictor for patients with gastric carcinoma by both multivariate Cox regression analysis (P = 0.021) and receiver operating characteristic curves (P = 0.001).CONCLUSION: CD73 expression correlates closely with HIF-1α expression in gastric carcinoma. CD73 could be an independent prognostic indicator for gastric carcinoma.     

Bmal1在胃癌组织中的表达及其临床意义

目的 探讨钟基冈Bmal1在胃癌和癌旁组织中的表达及其临床意义.方法 收集15例胃癌和癌旁非癌新鲜手术标本,以实时荧光定量PCR检测Bmall mRNA表达.收集64例胃癌和癌旁组织,以免疫组化方法检测Bmal1表达,用图像分析技术分析Bmal1在不同组织中的表达差异,结合临床资料探讨其与胃癌临床病理的相关性.结果 胃癌组织中Bmal1 mRNA转录量为1.02±0.05,癌旁组织中为0.57-0.03,二者比较差异有统计学意义(P＜0.05).Bmal1蛋白在癌旁组织中主要位于细胞的胞核,平均吸光度值为0.1041 ±0.0543,而在胃癌组织中主要位于胞质,平均吸光度值0.1816±0.0128,Bmall在胃癌组织与癌旁组织差异有统计学意义(P＜0.05).Bmal1表达强度与年龄、胃癌分化程度相关(P＜0.05),Bmal1与性别、肿瘤大小、浸润深度、淋巴结转移无相关性(P＞0.05).结论 Bmal1蛋白在胃癌组织中表达移位和表达上调可能与胃癌发生、发展相关,可能成为影响胃癌预后的重要因素.