DNA修复酶(MGMT)在人胃肠癌中表达的临床意义

目的探讨MGMT在胃肠癌中表达的临床意义。方法93例胃肠癌(其中胃癌53例,大肠癌40例)利用SABC免疫组织化学方法检测MGMT。结果在胃肠癌中MGMT阳性率为49.46%(46/93),在胃癌和大肠癌中阳性率分别为45.28%(24/53)和55.00%(22/40)。66岁以上老年组MGMT阳性率明显低于其他年龄组(P<0.05),癌组织浸润深度达浆膜组和有淋巴结转移组MGMT阳性率高。MGMT在胃肠癌中的表达与性别和癌组织类型、分化程度相关性不明显(P>0.05)。结论MGMT在胃肠癌中的表达与年龄、浸润深度和淋巴结转移有关。     

P-gp、HSP27和TopoⅡ在消化道癌表达的研究

目的探讨消化道癌组织中P-糖蛋白(P-gp)、热休克蛋白(HSP27)和拓扑异构酶(TopoⅡ)表达及意义.方法收集163例消化道癌标本,包括食道癌43例,胃癌48例,大肠癌72例,采用石蜡切片和免疫组织化学SP法染色.结果 P-gp在消化道癌中的阳性率为49.69%,HSP27在消化道癌中的阳性率为57.67%,TopoⅡ阳性率为47.85%.P-gp和HSP27在高分化组阳性率高于低分化组(P<0.05),而TopoⅡ消化道癌低分化组阳性率高于高分化组(P<0.05).TopoⅡ在食道癌组阳性表达率高于胃癌(P<0.05);而P-gp在大肠癌中的表达率明显高于胃癌(P<0.05).结论三种耐药标志物与癌组织分化程度相关;联合检测消化道癌组织中P-gp、HSP27和TopoⅡ,对制定化疗方案有指导作用.     

原发性胃癌REG 1A mRNA的表达与临床病理特性的关系研究

本文采用RT-PCR的方法检测胃癌组织的REG 1A基因mRNA表达.并分析其与胃癌临床特征的相关性.结果显示有78%(183/235)的原发性胃癌REG ;A mRNA阳性.REG 1A mRNA在胃癌中的表达与肿瘤的浸润生长方式、印戒细胞癌及低分化胃腺癌关系密切.与REG lA mR.NA阳性的肿瘤患者相比,REG lA mRNA阴性高分化腺癌患者有更好的预后.同时REG 1A mR-NA阳性的肿瘤发生血行转移的机会要显著高于REG lA mRNA阴性肿瘤.因此REG lA mRNA的表达与胃癌浸润性生长密切相关,可能是高分化胃腺癌的一个不良预后指标.     

胃癌中CDC4和cyclin E的表达及临床意义

检测胃癌中CDC4/Fbxw7、cyclin E的表达,分析其与胃癌临床病理特征的关系和临床意义.采用逆转录多聚酶链反应(RT-PCR)检测部分胃癌和胃正常组织中CDC4、cyclin E mRNA的表达;免疫组化(SP法)检测60例胃癌组织及对应正常组织中CDC4/FBXW7、cyclin E蛋白的表达,探讨两者的临床病理特征之间的关系.胃癌中CDC4蛋白表达显著低于正常组织(P<0.05),而cyclin E蛋白的表达在胃癌组织中显著增高,阳性率与癌旁正常组织比较,差异有统计学意义(P<0.01).CDC4、cyclin E蛋白和mRNA表达均与胃癌的分化程度、TNM分期、淋巴结转移及浸润深度有关.CDC4、cyclin E在mRNA表达水平上呈负相关.CDC4的表达缺失可能导致cyclin E的过表达.CDC4的低表达可能是胃癌诊疗及预后判断的重要生物学指标.     

CD90过表达影响胃癌细胞AGS的干性特征

2019年中国癌症报告显示,胃癌发病率仅次于肺癌,位列第二,其死亡率排在所有肿瘤的第三位,严重危害人们健康.筛查和鉴定胃癌的早期检测标志物、寻找胃癌治疗的分子靶点,对于降低胃癌致死率至关重要. CD90 (THY1)是一种细胞表面糖蛋白,在肿瘤细胞增殖、转移以及血管生成中发挥重要作用. CD90异常表达与干细胞特性有关,促进肿瘤的起始与转移.但CD90与胃癌细胞干性特征之间的关系尚未见文献报道.本研究发现,CD90过表达增加胃癌细胞AGS的侧群细胞(SP细胞)比例,影响其膜表面ABCG2、CD105等干性标记物的蛋白质表达水平,同时也影响胃癌细胞AGS胞内NANOG、SOX2等干性标记物的m RNA表达水平,mRNA芯片和Western-blot技术揭示CD90过表达可能通过调控PI3K/Akt和JNK/ERK1/2信号通路活性,影响胃癌细胞AGS的干性特征.本实验结果为胃癌潜在分子靶点的鉴定提供了新的思路.     

胃癌survivin基因mRNA和蛋白的表达与临床病理关系

目的　研究胃癌组织中survivin基因的mRNA和蛋白表达情况及其与临床病理参数的关系。方法　应用原位杂交方法和免疫组化SP法检测 5 4例胃癌 ,34例胃良性病变 ,2 0例胃正常组织标本中survivin基因mRNA及蛋白的表达。并对其与临床病理因素和二者的关系进行分析。结果　Survivin蛋白表达阳性率在胃癌、良性疾病组织和正常组织分别为 87 0 % (47/ 5 4 )、 2 3 5 % (8/ 34)和 15 % (3/ 2 0 ) ;SurvivinmRNA阳性率为 79 6 % (43/ 5 4 )、 2 3 5 % (8/ 34)和2 0 % (4/ 2 0 )。胃癌组远大于正常胃组织和良性疾病组 ,而正常胃组织与胃良性疾病组之间差异无显著性。SurvivinmRNA与蛋白在胃癌中的表达呈正相关 (rs=0 6 79,P <0 0 5 )。且其阳性率高低与性别、年龄、组织学类型无关 ;而与淋巴结转移、TNM分期、组织学分级有关。结论　SurvivinmRNA与蛋白在胃癌中表达较高 ,且与淋巴结转移、TNM分期、组织学分级有关 ,它可作为评估胃癌生物学行为和判断预后的生物学指标。     

SRSF1和BIRC5在胃癌组织中的表达及临床意义

目的通过对丝氨酸/精氨酸富有剪接因子1(SRSF1)和凋亡抑制因子(BIRC5)在胃癌组织中表达的检测及两者之间相关性的研究,来探讨两者与胃癌的发生、发展之间的关系;为胃癌的转移及复发预警,以及预后评估提供新靶点。方法应用逆转录-聚合酶链式反应(RT-PCR)和蛋白质印迹(Western blotting)检测60例术中切除的胃癌肿组织以及对应的癌旁组织中SRSF1与BIRC5的表达水平,分析这两种蛋白的表达水平与临床病理学特征的关系。结果 (1)胃癌肿组织中SRSF1 mRNA和BIRC5mRNA表达情况:SRSF1阳性表达率(75.0%,45/60)高于癌旁组织中(8.3%,5/60);BIRC5阳性表达率(81.7%,49/60)高于癌旁组织中(11.7%,7/60)。胃癌肿组织中SRSF1mRNA与BIRC5mRNA的表达与患者的年龄、性别、肿瘤大小等因素无关,与胃癌肿组织的分化程度、淋巴结转移以及TNM分期有关;SRSF1和BIRC5 mRNA在胃癌肿组织中的阳性表达呈正相关(r=0.3496,P=0.0062)。(2)胃癌肿组织中SRSF1蛋白与BIRC5蛋白表达情况:胃癌肿组织中SRSF1蛋白表达量相较于其在癌旁组织中的表达量明显增多(t=44.87,P0.01),BIRC5蛋白表达量较癌旁组织同样异常增高(t=69.84,P0.01)。结论在胃癌肿组织中SRSF1和BIRC5表达均存在异常增高,与胃癌的发生发展存在正相关,这可能与SRSF1选择性剪接BIRC5片段,促进凋亡抑制基因BIRC5的高表达有关,具体机制还需进一步探讨。     

胃癌CD44v6 mRNA及其蛋白表达与临床病理特征的关系

目的探讨细胞粘附分子CD44v6 mRNA及其蛋白表达与胃癌临床病理学行为和患者预后的关系. 方法应用高敏感性催化信号放大系统(catalyzed signal amplification,CSA)原位杂交和免疫组化技术,对17例早期胃癌、21例中期胃癌和57例晚期胃癌组织进行CD44v6 mRNA及其蛋白检测,并结合肿瘤的病理学行为和临床随访资料进行分析.结果在胃癌中,CD44v6 mRNA及其蛋白的表达阳性率分别为85.3%和82.1%.CD44v6 mRNA及其蛋白表达阳性率在晚期胃癌明显高于早、中期胃癌(P<0.05).CD44v6 mRNA表达与蛋白表达水平具有一致性,均与胃癌浆膜浸润,淋巴结转移和患者预后呈正相关(P<0.05).结论 CD44v6 mRNA及其蛋白异常表达与胃癌的临床病理生物学行为密切相关,特别是与胃癌细胞的转移潜能和胃癌患者的不良预后密切相关.CD44v6蛋白水平的表达可以间接反映其mRNA转录水平,并可作为预测胃癌转移潜能和患者预后的一个新的生物学指标.     

胃癌组织中端粒酶活性表达的初步研究

目的 :端粒酶的激活与胃癌发生密切相关 ,通过检测胃癌组织中端粒酶活性表达水平 ,为胃癌及其发生的早期诊断提供一种有效的方法。方法 :应用PCR ELISA技术 ,把端粒酶重复序列扩增技术与微孔液相杂交酶联呈色技术有机结合 ,检测胃肠癌组织和腹水标本中的端粒酶活性表达。结果 :1 7例晚期胃癌组织标本中检出 1 3例端粒酶活性表达 ,阳性率为 76 47% ;2 3例中期胃肠癌组织标本中检出 1 9例端粒酶活性表达 ,阳性率为 78 2 6% ;66例早期胃癌组织标本中检出5 7例端粒酶活性表达 ,阳性率为 86 36% ;36例癌旁近端组织标本中检出 1 1例端粒酶活性表达 ,阳性率为 30 5 6% ;36例癌旁远端组织标本中检出 4例端粒酶活性表达 ,阳性率为 1 1 1 1 % ;2 6例胃肠癌腹水标本检出 1 9例端粒酶活性表达 ,阳性率为 73 0 8% ;5 4例正常人胸腹水标本中没有检出端粒酶活性表达。结论 :胃癌组织标本中有较高的端粒酶活性表达 ,端粒酶活性表达可以作为胃癌发生的有效指标 ,而PCR ELISA技术可以定量评价作为端粒酶活性表达从而作为诊断胃癌发生的一种快速简便有效的方法。     

热休克蛋白在高温致神经管畸形中的表达

目的：检测热休克蛋白在高温致神经管畸形中的表达状况,以探讨高温致神经管畸形的机制。方法：在高温致金黄地鼠神经管畸形的动物模型上,利用免疫组织化学(SABC法)方法,检测高温致神经管畸形中,热休克蛋白(HSP70和HSP90)在神经上皮细胞及周围间充质细胞中的表达状况;同时利用地高辛标记的寡核苷酸探针进行原位杂交,检测HSP70 mRNA和HSP90 mRNA在神经上皮细胞及周围间充质细胞中的转录状况。结果：高温处理后2h,鼠胚神经上皮细胞及周围间充质细胞HSP70、HSP90的表达与正常对照组相比明显增强,8h和16h的表达达到高峰,24h后与对照组水平一致。原位杂交结果显示,高温处理后2h神经上皮细胞及周围间充质细胞中出现HSP70 mRNA及HSP90mRNA杂交阳性信号,8h阳性信号最强,16h后阳性信号减弱,至24h后未见阳性信号。结论：高温可引起神经上皮细胞及其周围间充质细胞HSP70和HSP90应激性表达,这可能是胚胎受到高温作用后发生的一种保护性反应。     

High Expression of Heat Shock Protein 90 Is Associated with Tumor Aggressiveness and Poor Prognosis in Patients with Advanced Gastric Cancer

The heat shock protein 90 (HSP90) is overexpressed and highly associated with poor prognosis in many malignancies. However, the role of HSP90 in gastric cancer has not been thoroughly elucidated. The aim of this study is to investigate the relationship of HSP90 expression with clinicopathological parameters and prognosis in advanced gastric cancer, and estimate the alteration of HSP90 expression after neoadjuvant chemotherapy. HSP90 and matrix metallopeptidase 9 (MMP-9) antigen expression was evaluated by immunohistochemistry in 322 advanced gastric carcinoma samples. The relationships between HSP90 and clinicopathological parameters and prognosis were analyzed. The response of HSP90 level was assessed in chemotherapeutic effect in 54 patients received 1–2 cycles of neoadjuvant chemotherapy. The positive expression of HSP90 was found to be 69.6% in 322 advanced gastric carcinoma samples. HSP90 protein expression was significantly associated with depth invasion (P<0.001), lymph node metastasis (P<0.001) and stage of disease (P<0.001). The positive rates of HSP90 expression were higher in both prominent serosal invasion group (P<0.001) and lymph node metastasis group (P<0.001). Moreover, HSP90 was significantly correlated with MMP-9 among 322 gastric cancer tissues (P<0.001). In univariate and multivariate analyses, HSP90 was an independent prognostic factor for both recurrence-free survival (RFS) and overall survival (OS). These results suggested that HSP90 may play an important role on tumor invasion, metastasis and prognosis, and might act as a promising target for prognostic prediction.     

Transient changes of enzyme activities and expression of stress proteins in the small intestine of piglets after weaning

To determine the transient effects of weaning on the small intestine, 16 piglets were slaughtered at days 0, 1, 4 and 7 after weaning. Jejunal samples were collected to examine different enzyme activities and mRNA expressions of two stress protein families, namely, heat-shock proteins (HSP) and trefoil factors (TFF). Results showed that the activities of ceruloplasmin, alkaline phosphatase and lactate dehydrogenase, were significantly changed at Day 1 and/or Day 4. The mRNA expressions of HSP10, HSP60 and HSP90 showed a pattern of increased expression with time after weaning. Expression significantly differed between Day 0 and Day 7 after weaning. The mRNA expression of HSP70 was significantly increased on Day 1 only. Similarly, the mRNA expressions of TFF1 and TFF2 were significantly increased on Day 7 compared with those on Day 0. Expression of TFF3 was not affected by time after weaning. In conclusion, the present study indicated that weaning induced transient injury to small intestinal morphology and function. Particularly it changed enzyme activities and gene expression of stress proteins in the small intestine of piglets. At first time, a change in the gene expression of HSP10 and a gene overexpression of TFF1 in the small intestine of piglets after weaning was found.     

下调UCA1通过靶向miR-23b及其下游靶基因抑制胃癌细胞的增殖和转移

摘要 目的：探讨长链非编码RNA尿路上皮癌相关基因1（UCA1）调控胃癌细胞增殖和转移的分子机制。方法：将人胃癌细胞株SGC7901分为：对照组、siRNA-NC组、siRNA-UCA1组、inhibitor-NC组和miR-inhibitor组、si-UCA1+inhibitor-NC组和si-UCA1+miR-inhibitor组。对SGC7901细胞分别转染siRNA-UCA1及阴性对照（siRNA-NC）、miR-inhibitor及阴性对照（inhibitor-NC）,未转染的细胞作为对照组。通过RT-qPCR检测细胞中UCA1和miR-23b-3p的水平。通过CCK-8法、伤口愈合实验和Transwell实验评价细胞的增殖、迁移和侵袭能力。通过Western blot分析细胞中IL6R、BCL2和HSP90B1蛋白的表达。使用pcDNA-UCA1/pcDNA-NC与pGL3-miR-23b-3p-WT/pGL3-miR-23b-3p-Mut共转染细胞,通过双荧光素酶报告实验验证UCA1与miR-23b-3p的靶向关系。结果：细胞培养48 h和72 h后,与对照组比较,siRNA-UCA1组的细胞活力分别降低了31.58%和31.40%（P<0.05）。与对照组比较,siRNA-UCA1组的细胞迁移率[(61.46±5.43)% vs (23.16±3.17)%]、侵袭细胞数量（109.17±9.66 vs 50.83±6.96）、IL6R、BCL2和HSP90B1的蛋白相对表达量均显著降低,而miR-23b-3p相对表达量升高（P<0.05）。与pGL3-miR-23b-3p-WT共转染后,与pcDNA-NC组比较,pcDNA-UCA1组的相对荧光酶活性降低了66.12%（P<0.05）。与si-UCA1+inhibitor-NC组比较,si-UCA1+miR-inhibitor组的细胞活力、细胞迁移率、侵袭细胞数量、IL6R、BCL2和HSP90B1的蛋白相对表达量均显著升高（P<0.05）。结论：下调UCA1通过靶向miR-23b-3p及其下游基因IL6R、BCL2和HSP90B1来抑制胃癌细胞的增殖和转移。     

The role of HSP90 molecular chaperones in hepatocellular carcinoma

Misfolded proteins have enhanced formation of toxic oligomers and nonfunctional protein copies lead to recruiting wild-type protein types. Heat shock protein 90 (HSP90) is a molecular chaperone generated by cells that are involved in many cellular functions through regulation of folding and/or localization of large multi-protein complexes as well as client proteins. HSP90 can regulate a number of different cellular processes including cell proliferation, motility, angiogenesis, signal transduction, and adaptation to stress. HSP90 makes the mutated oncoproteins able to avoid misfolding and degradation and permits the malignant transformation. As a result, HSP90 is an important factor in several signaling pathways associated with tumorigenicity, therapy resistance, and inhibiting apoptosis. Clinically, the upregulation of HSP90 expression in hepatocellular carcinoma (HCC) is linked with advanced stages and inappropriate survival in cases suffering from this kind of cancer. The present review comprehensively assesses HSP90 functions and its possible usefulness as a potential diagnostic biomarker and therapeutic option for HCC.     

应用蛋白质组学技术筛选胃癌耐药相关蛋白质

胃癌多药耐药性是临床胃癌化疗失败最主要的原因之一,但其分子机制仍然不太清楚.为了寻找新的胃癌耐药相关的蛋白质,揭示胃癌多药耐药的分子机制,以胃癌细胞SGC7901和长春新碱诱导的耐药胃癌细胞SGC7901/VCR为研究对象,应用二维凝胶电泳(two-dimensionalelectrophoresis,2-DE)技术分离两种细胞的总蛋白质,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(matrix-assistedlaserdesorption/ionizationtimeofflightmassspectrometry,MALDI-TOF-MS)及电喷雾电离串联质谱(electrosprayionizationtandemmassspectrometry,ESI-Q-TOF)对差异表达的蛋白质点进行鉴定,蛋白质印迹和实时RT-PCR验证部分差异蛋白质在两株细胞中的表达水平,反义核酸转染技术分析HSP27(heatshockprotein27,HSP27)高表达与SGC7901/VCR耐药的相关性.得到了分辨率较高、重复性较好的两株细胞系的二维凝胶电泳图谱,质谱分析共鉴定了24个差异蛋白质点,蛋白质印迹和实时RT-PCR验证了部分差异蛋白的表达水平,反义寡核苷酸抑制HSP27表达能增加SGC7901/VCR对长春新碱的敏感性.研究结果不仅提示这些差异蛋白质如HSP27,Sorcin等可能与胃癌的多药耐药相关,而且为揭示胃癌细胞的多药耐药性产生机制提供了线索.     

Cloning HSP70 and HSP90 genes of kaluga (<Emphasis Type="Italic">Huso dauricus</Emphasis>) and the effects of temperature and salinity stress on their gene expression

The genes encoding HSP70 and HSP90 proteins were isolated from kaluga by homologous cloning and rapid amplification of complementary DNA (cDNA) ends (RACE). HSP70 (GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"KP050541","term_id":"756558159","term_text":"KP050541"}}KP050541) and HSP90 (GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"KP050542","term_id":"756558161","term_text":"KP050542"}}KP050542) cDNAs were composed of 2275 and 2718 bp and encoded polypeptides of 650 and 725 amino acids, respectively. Basic Local Alignment Search Tool (BLAST) analysis showed that HSP70 and HSP90 of kaluga shared high identities with those of Acipenser ruthenus, Acipenser schrenckii, and Acipenser baerii (98–99 %). Fluorescent real-time RT-PCR under unstressed conditions revealed that HSP70 and HSP90 were expressed in 11 different tissues of kaluga. Messenger RNA (mRNA) expressions of both HSP70 and HSP90 were highest in the intestine and lowest in the muscle. In addition, the patterns of mRNA expression of HSP70 and HSP90 were similar, although the level of expression was more in HSP90 than in HSP70 (P < 0.05).We also analyzed patterns of HSP70 and HSP90 expression in the muscle, gill, and liver of kaluga under different combinations of temperature and salinity stress, including temperatures of 4,10, 25, and 28 °C at 0 ppt salinity, and salinities of 10, 20, 30, and 40 ppt at 16 °C, where 16 °C at 0 ppt (parts per thousand) served as the control. We found that levels of mRNA expression of both HSP70 and HSP90 were highest at 4 °C in the muscle, gill, and liver and changed little with salinity stress. These results increase understanding of the mechanisms of stress response of cold freshwater fish.     

胃癌nm23H1 mRNA、VEGF-C mRNA表达及其与淋巴转移、生存率的相关性研究

研究胃癌组织nm23H1 mRNA、VEGF-C mRNA和VEGFR-3的表达与胃癌中的新生淋巴管的生成、肿瘤淋巴道转移和生存率关系,为临床治疗及预后提供实验依据。用胃癌与癌周组织作对照,应用原位杂交法检测78例胃癌组织nm23H1 mRNA、VEGF-C mRNA表达,并以VEGFR-3抗体为标记,经EnVisionTM免疫组织化学及Leica-Qwin计算机图像分析,用Weidner最高血管密度计数法,计数阳性淋巴管数(MLC),以及调查患者术后的生存率。胃癌nm23H1 mRNA阳性表达 69．23％(54例)、nm23H1 mRNA阳性表达与胃癌淋巴结转移、TNM分期、MLC呈负相关(P<0．01), VEGF-C mRNA阳性表达46．15％(36例),其与胃癌淋巴结转移、TNM分期、MLC呈正相关(P<0．01 或P<0．05)．与癌周组织的nm23H1 mRNA和VEGF-C mRNA表达比较,具有显著性差异(P<0．01 或P<0．05)。nm23H1 mRNA在Ⅰ、Ⅱ期胃癌中呈高表达,在Ⅲ、Ⅳ期患者中呈低或无表达。胃癌组织的MLC(8．37±2．29/mm2)显著高于癌周组织(4．51±2．64/mm2),两者比较具有显著性差异(P<0．05)。 nm23H1 mRNA与VEGFR-3表达之间存在负相关(P<0．05,r=0．8479);VEGF-C mRNA与VEG- FR-3表达之间存在正相关(P<0．05,r=0．8362)。在根治术5年内死亡的61．54％(48例)患者中 MLC(10.82±2．51/mm2)高于5年内生存的患者(6．53±2．09/mm2),两组间有显著性差异(P<0．05);在不同的肿瘤病理学分级中,胃癌未、低分化型与高、中分化型的nm23H1 mRNA阳性表达,有显著性差异(P<0．05),当nm23H1 mRNA高表达时,肿瘤淋巴转移率低,生存率高,故认为nm23H1基因具有抑制胃癌发生和淋巴道转移作用;当VEGF-C mRNA高表达时,肿瘤淋巴转移率高,生存率低。胃癌组织中的VEGFR-3表达水平与肿瘤的淋巴转移密切相关,胃癌组织MLC的显著增高,提示肿瘤组织有新淋巴管的生成。VEGF-C促进了肿瘤诱导的淋巴管新生,在胃癌的淋巴道转移中起重要作用。