CHARACTERISTICS OF MUCINS AND CEA IN GLOBOID DYSPLASTIC CELLS OF HUMAN STOMACH AND ITS RELATIONSHIP WITH SIGNET RING CELL CARCINOMA

Fourty-three cases of globoid dysplasia and signet ring cell carcinoma were stained by mucin and CEA (ABC method). It was found that there were three kinds of mucins (neutral, sialo and sulphomucin in both globoid dysplastic cells and signet ring cells. The percentages of the three kinds of mucins seen in the two kinds of cells were not much different. It was indicated that the altered mucins in the gastric epithelial cells must be a sign of dedifferentiation of the cells and the results of malfunction. The globoid dysplasia type I mainly contained neutral mucin, whereas type II, the acid mucin was predominant, especially the sulphomucin. The CEA positive reaction became stronger as the atypia being remarkable, and the characteristics of distribution of CEA positive particles were similar in the two kinds of cells namely, randomly or disorderly in the cells. Based on the analysis of the results, a conclusion can be made that the variety of mucins in globoid dysplastic cells can be used as a reference point in classification and is not much significant in grading, but the amount of CEA positive matter can be a reference point in grading. The globoid dysplasia is such a lesion with special features in morphology and function manifested in the process of de-differentiation towards signet ring cell carcinoma following the successive action of carcinogens upon the cells of gastric epithelium.

Neoadjuvant chemotherapy vs upfront surgery for gastric signet ring cell carcinoma:A retrospective,propensity score-matched study

BACKGROUND The benefit of neoadjuvant chemotherapy for patients with signet-ring cell carcinoma of the stomach is controversial.AIM To evaluate the perioperative and long-term outcomes of neoadjuvant chemotherapy for locally advanced gastric signet-ring cell carcinoma.METHODS This retrospective study identified patients with locally advanced signet-ring cell carcinomas of the stomach(cT3/4 and cN any)diagnosed from January 2012 to December 2017 by using the clinical Tumor-Node-Metastasis(cTNM)staging system.We performed 1:1 propensity score matching(PSM)to reduce bias in patient selection.The histologic and prognostic effects of neoadjuvant chemotherapy were assessed.The overall survival rates were used as the outcome measure to compare the efficacy of neoadjuvant chemotherapy vs surgery-first treatment in the selected patients.RESULTS Of the 144 patients eligible for this study,36 received neoadjuvant chemotherapy,and 108 received initial surgery after diagnosis.After adjustment by PSM,36 pairs of patients were generated,and baseline characteristics,including age,sex,American Society of Anesthesiologists score,tumor location,and cTNM stage,were similar between the two groups.The R0 resection rates were 88.9%and 86.1%in the surgery-first and neoadjuvant chemotherapy groups after PSM,respectively(P=1.000).The median follow-up period was 46.4 mo.The 5-year overall survival rates of the neoadjuvant chemotherapy group and surgery-first group were 50.0%and 65.0%(P=0.235),respectively,before PSM and 50%and 64.7%(P=0.192),respectively,after PSM.Multivariate analyses conducted before and after PSM showed that NAC was not a prognostic factor.CONCLUSION Neoadjuvant chemotherapy provides no survival benefit in patients with locally advanced gastric signet-ring cell carcinoma.For resectable gastric signet-ring cell carcinoma,upfront surgery should be the primary therapy.

EXPRESSION OF PCNA,AKP AND ACP DURING DEVELOPMENT OF MOUSE FORE STOMACH CARCINOMA

Objective： To find out the relationship of the expressions of proliferating cell nuclear antigen （proliferating cell nuclear antigen, PCNA）, alkaline phosphotase （alkaline phosphotase, AKP） and acid phosphotase （acid phosphotase, ACP） with the development of mouse fore stomach cancerization. Methods： The animal models, including the various stages during the development of NIH mouse fore stomach carcinoma, were made by N-Nitrososarcosineethylester （N-Nitrososarcosineethylester, NSEE）. The mice were sacrificed on the 14th, 28th, 42nd, 56th, 70th and 84th days respectively after mice were irrigated with NSEE. The fore stomach was taken out and dissected. The methods of histopathology, immunohistochemistry and isoenzyme electrophoresis were adopted to study the dynamic changes of cell shape and expression of PCNA, AKP and ACP. Results： On the 42nd and 56th days after NSEE treatment, the expression of PCNA increased gradually along with the cancerization. Comparing with the control, there were significant differences （P〈0.05）. On the 70th and 84th days, the expression of PCNA increased further （compared with the control P〈0.01）. The activity of AKP increased gradually along with the cancerization. On the 14th, 28th, 42nd and 56th days, there were significant differences （P〈0.05）; on the 70th and 84th days, the activity of AKP increased further （P〈0.01）. The activity of ACP also increased on the 14th, 28th, 42nd and 56th days, and there were significant differences on the 70th days （P〈0.05） and on the 84th days （P〈0.01） compared with the control. Conclusion： During the carcinogenesis of NIH mouse fore stomach, the expressions of PCNA, AKP and ACP increased gradually and were consisted with the changes of cell shapes.

Killing effect of TNF-related apoptosis inducing ligand regulated by tetracycline on gastric cancer cell line NCI-N87

AIM: To clone the cDNA fragment of human TRAIL (TNF-related apoptosis inducing ligand) into a tetracycline-regulated gene expression system, the RevTet-On system, transduce expression vectors into a gastric carcinoma cell line-NCI-N87 and examine the effects of controlled expression of TRAIL in vitro on the gastric carcinoma cells. METHODS: The full-length cDNA of TRAIL was inserted into a vector under the control of the tetracycline-responsive element (TRE) to obtain the plasmid pRevTRE-TRAIL, which was transfected into a packaging cell line PT67. In addition, vector pRev-Tet On and pRevTRE were also transfected into PT67 separately. After hygromycin and G418 selection, the viral titer was determined. The medium containing retroviral vectors was collected and used to transduce a gastric carcinoma cell line NCI-N87. The resulting cell line NCI-N87-Tet On TRE-TRAIL and a control cell line, NCI-N87 Tet On-TRE, were established. TRAIL expression in the cell line was induced by incubating cells with doxycycline (Dox), which is a tetracycline analogue. The killing effect on gastric carcinoma cells was analyzed after induction. RESULTS: The recombinant plasmid pRev-TRE-TRAIL was constructed. After hygromycin or G418 selection, the producer cell lines PT67-TRE, PT67-TRE-TRAIL and PT67-Tet On were obtained,with titers of about 10(8)CFU.L(-1). By transducing NCI-N87 cells with retroviral vectors from these cell lines, stable cell lines NCI-N87-Tet-On TRE-TRAIL (NN3T) and control cell line NCI-N87-Tet-On-TRE (NN2T) were established. The growth curves of the selected cell lines were the same with the wild type NCI-N87. When Dox was added, cell death was obvious in the test groups (29%-77%), whereas no difference was observed in control and wild type cell lines. With the addition of a medium from the test group, human leukemia cell line Jurkat was activated till death (83%), indicating the secretion of active TRAIL proteins from the test cells to the medium. CONCLUSION: With the use of the RevTet-On system, a regulated expression system for TRAIL was constructed. Using this system, the selected killing effect of TRAIL on gastric carcinoma cell line NCI-N87 could be observed.

Blockade of cholecystokinin-2 receptor and cyclooxygenase-2 synergistically induces cell apoptosis, and inhibits the proliferation of human gastric cancer cells in vitro

Gastrin and cyclooxygenase-2(COX-2) playimportant roles in the carcinogenesis and progression ofgastric cancer.However,it remains unknown whether the combination of cholecystokinin-2(CCK-2) receptor antagonist plus COX-2 inhibitor exerts synergistic anti-tumor effects on human gastric cancer.Here,we demonstrated that the combination of AG-041R(a CCK-2 receptor antagonist) plus NS-398(a selective COX-2 inhibitor) treatment had synergistic effects on proliferation inhibition,apoptosis induction,down-regulation of Bcl-2 and up-regulation of Bax expression in MKN-45 cells.These results indicate that simultaneous targeting of CCK-2 receptor and COX-2 may inhibit gastric cancer development more effectively than targeting either molecule alone.(C)2008 Elsevier Ireland Ltd.All rights reserved.

CT动态容积灌注成像检出早期胃癌并评估其病理分型

目的 观察CT动态容积灌注(DVPCT)成像检出早期胃癌并鉴别其病理分型的价值。方法 回顾性分析127例经病理证实的早期胃癌患者,根据术前检查方式分为增强CT组(n=67)或DVPCT组(n=60);比较组间一般资料、CT资料,以及DVPCT组内胃印戒细胞癌(SRCC)与胃腺癌的强化程度、门静脉期与延迟期峰值期相及峰值时间;绘制受试者工作特征(ROC)曲线,计算曲线下面积,评估DVPCT时间-密度曲线(TDC)鉴别早期SRCC与腺癌的效能。结果 DVPCT组肿瘤检出率、剂量长度乘积及有效剂量均高于增强CT组(P均<0.05);2组患者年龄、性别、病理分型、肿瘤位置及肿瘤最大径差异均无统计学意义(P均>0.05)。52例(52/60,86.67%)经DVPCT检出早期胃癌的患者中,SRCC 12例、腺癌39例、黏液腺癌1例;其中,早期胃SRCC与早期胃腺癌患者肿瘤强化程度、门静脉期及延迟期峰值期相及峰值时间差异均有统计学意义(P均<0.05)。以峰值时间37.3 s为最佳截断值,DVPCT TDC鉴别早期胃SRCC与早期胃腺癌的敏感度、特异度、阳性预测值、阴性预测值、准确率及曲线下面积分别为83.33%、84.62%、62.50%、94.29%、84.31%及0.895。结论 DVPCT检出早期胃癌效果优于常规增强CT;TDC可有效鉴别早期胃SRCC与早期胃腺癌。

芒柄花黄素对人胃癌细胞株MKN-45增殖、凋亡的影响及其机制

目的观察芒柄花黄素对人胃癌MKN-45细胞株增殖、凋亡的影响,并探讨其可能作用机制。方法取对数生长期人胃癌MKN-45细胞分为A、B、C、对照组,A、B、C组分别加入20、40、80μg/m L芒柄花黄素培养,对照组加入不含芒柄花黄素的完全培养基。采用MTT法观察给药24、48、72 h各组细胞增殖情况,计算细胞增殖抑制率。采用DAPI染色法评价其对MKN-45细胞形态学的影响。采用Western blotting法检测给药48 h时各组细胞磷酸化IκB(p-IκB)及NF-κB p65。结果随芒柄花黄素浓度升高,细胞增殖抑制率升高,且随干预时间增加,细胞增殖抑制率升高(P均<0.05)。给药72 h时A、B、C、对照组的凋亡率分别为25.62%±2.57%、48.27%±3.18%、72.51%±4.35%、1.07%±0.54%,A、B、C组与对照组相比,P均<0.05。给药48 h时A、B、C组细胞p-IκB、NF-κB p65蛋白相对表达量均高于对照组,且随芒柄花黄素浓度升高,细胞p-IκB及NF-κB p65蛋白相对表达量增加,P均<0.05。结论芒柄花黄素可抑制人胃癌MKN-45细胞株的增殖、促进细胞凋亡,其机制可能为芒柄花黄素激活NF-κB信号通路。

CD4^+CD25^+调节性T细胞在胃癌患者的表达及其临床意义

目的研究胃癌患者肿瘤组织、淋巴结及外周血中CD4+、CD25+调节性T细胞(Tregulatorycells,Treg)的表达情况与胃癌临床病理的关系并分析其意义。方法胃癌患者肿瘤组织、淋巴结、外周血及对照组中Treg表达情况分别以冰冻切片荧光抗体染色、流式细胞仪进行检测,结合胃癌患者临床病理情况分析。结果健康成人对照外周血Treg表达率为(4.1±5.7)%,胃癌患者外周血Treg表达显著增高(16.8±7.9)%。胃癌组织Treg表达为(6.5±7.2)%,胃癌周围淋巴结Treg比例为(8.4±7.2)%,均较对照组显著增高。胃癌外周血和淋巴结Treg表达与淋巴结转移率和TNM分期有着负相关性,淋巴结转移阳性和TNM分期越晚,Treg表达率越高。结论Treg在胃癌患者的瘤组织、淋巴结及外周血中表达较对照组显著增高,并与临床病理存在显著相关性。

胃小细胞癌5例临床病理观察

目的观察胃小细胞癌(SCC)的临床病理特点。方法对5例胃SCC的组织切片及免疫组化结果进行观察并结合临床资料及术后随访结果进行分析。结果5例胃SCC患者均为男性,平均年龄57.6岁。主要临床表现为黑便、上腹痛、吞咽困难和消瘦,肿瘤分别位于胃体部、窦部、贲门和残胃吻合口。胃SCC肿瘤主要形成巢片状结构,瘤细胞大小及形态一致,核呈卵圆形、短梭形或不规则形,有异型,染色质为粗细不一的颗粒状,部分深染,核分裂象平均44个/10HPF;4例有片状坏死;2例可见腺样分化并各自伴有类癌或腺癌及印戒细胞癌。各例肿瘤呈明显的浸润性生长并伴有局部淋巴结转移,1例有肝转移。免疫组化示4例SCCSyn、CK8、CKAE1/AE3及NSE(+)。随访4例,3例死于肿瘤,平均生存8.3个月,1例已存活25个月,该例无肿瘤坏死。结论胃SCC是一种组织形态和生物学行为与肺SCC相似的高度恶性肿瘤,对Syn和CK8的表达是其免疫表型特点,肿瘤性坏死是评价预后的重要指标。

胃神经内分泌癌和胃癌伴神经内分泌分化的临床病理研究

目的 :探讨胃神经内分泌癌和胃癌伴神经内分泌分化的临床病理特征。 方法 :回顾 12例胃神经内分泌癌和 8例胃癌伴神经内分泌分化的临床病理资料及苏木精 伊红染色切片 ,并作神经内分泌标记物 (CgA、syn、NSE等 )的免疫组化染色和AB/PAS组化染色。 结果 :两组患者均以老年男性为主 ,肿块大多为溃疡型 ,多位于胃贲门部或胃窦部。 12例胃神经内分泌癌中 ,典型类癌 2例、不典型类癌 9例、低分化神经内分泌癌 1例 ,大部分癌细胞均表达神经内分泌标记物 ,其中 7例不典型类癌内见少数腺癌或粘液细胞癌成分 ,后二者阳性表达CEA和PAS。8例胃癌伴神经内分泌分化者见少数神经内分泌标记物阳性的细胞散在分布于胃癌组织内。 结论 :胃神经内分泌癌和胃癌伴神经内分泌分化属不同的组织学类型 。

胃印戒细胞癌的临床病理特征及耐药基因表达特点分析

目的探讨胃印戒细胞癌的病理学及P-糖蛋白(P-gp)、谷胱甘肽转移酶(GST-π)、拓扑异构酶Ⅱ(TopoⅡ)的表达特点,进一步提高其诊治水平。方法分析32例胃印戒细胞癌(胃镜活检标本21例,大部切除胃标本11例)的临床病理及GST-π、P-gp、TopoⅡ的表达特点。结果胃镜下以经典型印戒细胞数量最多,弥漫性生长;相关基因检测结果显示,GST-π、P-gp、TopoⅡ阳性表达率与同期32例高—中分化腺癌比较,GST-π阳性率无显著差异,P-gp阳性率明显升高,TopoⅡ阳性率明显降低(P均<0.05)。结论胃印戒细胞癌以典型印戒细胞数量最多,且呈弥漫性生长;GST-π、P-gp、TopoⅡ的表达检测有助于胃印戒细胞癌诊断。

膜联蛋白A7低表达对胃癌MGC-803细胞增殖和凋亡的影响

目的:探讨RNA干扰技术靶向沉默膜联蛋白A7(anexin A7,ANXA7)的表达对人胃癌MGC-803细胞增殖和凋亡的影响。方法:实验分为3组,以靶向ANXA7的siRNA转染MGC-803细胞为siRNA干扰组,另设阴性siRNA对照和空白对照组。Western blotting检测转染后MGC-803细胞中ANXA7蛋白的表达,应用MTT、集落形成和流式细胞术检测ANXA7下调对MGC-803细胞增殖、凋亡的影响。结果:靶向ANXA7的siRNA转染后MGC-803细胞中ANXA7的表达较阴性对照和空白对照组细胞明显降低[(21.79±1.72)%vs(99.87±5.13)%、(93.45±4.89)%,均P<0.05]。下调ANXA7明显降低MGC-803细胞的增殖能力[(0.97±0.06)vs(1.21±0.07)、(1.25±0.08),均P<0.05]和集落形成能力[(183±21)vs(363±35)、(348±27)个,均P<0.05],细胞凋亡率无显著变化(P>0.05)。结论:ANXA7低表达可抑制胃癌细胞MGC-803增殖,而对细胞凋亡无明显影响。

胃印戒细胞癌和癌旁球样型增生细胞DNA含量的研究

目的：了解球样异型增生细胞的癌前意义。方法：应用ＣＭＩＡＳ－００７型图象分析系统检测３４例胃印戒细胞癌、４４例球样异型增生细胞和１５例正常胃粘膜上皮细胞的ＤＮＡ含量。结果：胃印戒细胞癌、球样异型增生细胞和正常胃粘膜之间ＤＮＡ含量差异有显著性（Ｐ＜０．０５及Ｐ＜０．０１）。直方图上，正常胃粘膜细胞呈位于２Ｃ区的单峰型。胃印戒细胞癌的众数峰右移，分布分散，各例都有＞５Ｃ的非整倍体细胞出现。球样异型增生细胞的图象介于两者之间，其中Ⅲ级球样异型增生细胞含有＞５Ｃ的非整倍体细胞。结论：球样异型增生细胞是一种异常增生细胞，是胃印戒细胞癌的癌前病变。

人胃癌肿瘤相关成纤维细胞的原代培养和鉴定

目的探讨人胃癌肿瘤相关成纤维细胞(carcinoma-associated fibroblasts,CAFs)的培养和鉴定方法,并分析其与胃正常成纤维细胞(normal fibroblasts,NFs)在生物学特性和蛋白表达方面的差异,为其功能研究奠定基础。方法采用组织块法和酶消化法进行人胃癌CAFs和NFs的原代培养,通过酶消化法进行人工纯化,通过形态学观察和多种蛋白的免疫细胞化学染色对其进行鉴定。结果获得纯化的人胃癌CAFs和NFs。胃癌CAFs呈长梭形,大小不等,生长密集,排列紊乱。除表达vimen-tin这一成纤维细胞的标记物外,还表达FAP、SMA,但不表达CK、desmin。结论只要给细胞创造良好的生长条件并规范操作,组织块法和消化法均可成功的培养出人胃癌CAFs和人胃NFs。胃癌CAFs与NFs在形态结构、生长方式和蛋白表达等方面均存在明显差异。

干细胞相关基因Nestin在胃神经内分泌肿瘤中的表达及临床意义

目的研究干细胞相关基因Nestin在胃神经内分泌肿瘤中的表达,分析其与肿瘤生物学特点及患者生存时间的关系。方法应用组织芯片及免疫组织化学法,检测91例胃神经内分泌肿瘤标本及正常组织中Nestin的表达,并结合胃神经内分泌肿瘤不同病理亚型、临床病理参数和患者预后进行分析。结果 91例中,27例(29.67%)胃神经内分泌癌表达Nestin,而胃神经内分泌瘤及正常胃黏膜组织均不表达Nestin。Nestin表达与胃神经内分泌癌患者的性别、肿瘤浸润深度、脉管侵犯及TNM分期有显著相关性。Kaplan-Meier曲线显示,Nestin阳性表达的胃神经内分泌癌患者生存率显著低于阴性表达组。结论 Nestin在胃神经内分泌肿瘤不同的病理亚型间差别表达,Nestin的表达可能促进了胃神经内分泌癌的发生发展,并可能作为胃神经内分泌癌的干细胞标记物用于判断预后。

胃印戒细胞癌患者的临床及病理学特征

目的:探讨胃印戒细胞癌特殊的临床病理学特点,以指导临床早期发现和治疗。方法:总结15例手术后经病理证实为胃印戒细胞癌的临床及病理学特征。结果:15例患者发生网膜转移1例,淋巴结转移10例;发病部位以胃窦部小弯侧多见(7例);病变类型多为溃疡型(9例);术前4例患者出现血癌胚抗原(CEA)水平升高。结论:胃印戒细胞癌是弥漫型浸润性生长的肿瘤,发现时常浸润全层,多伴有淋巴结转移,血CEA水平增高对早期诊断起到一定的预警作用。

RXRγ、RARβ在9-cis-RA抑制人胃癌SGC7901细胞生长中的作用

目的:探讨维甲类X受体(RXR)γ、RARβ在RXR激动剂9-顺维甲酸(9-cis-RA)抑制人胃癌SGC7901细胞生长中的作用。方法:体外培养SGC7901细胞给予9-cis-RA干预,四甲基偶氮唑蓝(MTT)法、流式细胞术、HE染色、免疫组化染色、Western-blot检测9-cis-RA作用后SGC7901细胞生长情况、凋亡率、细胞周期的改变、RXRγ及RARβ表达情况。结果:SGC7901细胞与9-cis-RA共同培养48h后,肿瘤细胞生长受到抑制,其作用具有浓度及时间依赖性。流式细胞仪检测显示处于G0/G1期的细胞增多,凋亡率增高,免疫组化及Western-blot检测显示20μmol/L的9-cis-RA作用72h后,RXRγ、RARβ蛋白表达增加。结论:9-cis-RA可通过上调RXRγ、RARβ蛋白表达诱导细胞凋亡从而抑制人胃癌SGC7901细胞生长。

CD44v6在胃神经内分泌癌中的表达及其意义

目的探讨肿瘤干细胞表面标记物CD44v6在胃神经内分泌癌中的表达及其意义。方法免疫组织化学方法检测50例胃神经内分泌癌组织和相应癌旁组织中CD44v6蛋白的表达情况,分析CD44v6与胃神经内分泌癌临床病理特征及预后之间的关系。结果 CD44v6在胃神经内分泌癌中的表达率为52.0%,在癌旁组织中未见表达,两者差别有统计学意义(P<0.05)。CD44v6与浸润深度、N分期、TNM分期和细胞类型相关。CD44v6表达阳性与表达阴性患者术后5年生存率分别为19.4%和54.2%,差别有统计学意义(P<0.05)。单因素分析提示,浸润深度、N分期、TNM分期、CD44v6高表达是影响患者预后的相关因素;多因素分析提示,CD44v6高表达和N分期为胃神经内分泌癌的独立预后因素。结论 CD44v6参与胃神经内分泌癌浸润和淋巴结转移过程,可能是治疗胃神经内分泌癌的潜在靶点之一。

原发性胃鳞状细胞癌3例及文献复习

原发性胃鳞状细胞癌临床罕见,多为胃上部受累,>60岁男性多见,国内外共报道约百余例.病因及发病机制不详,较之胃腺癌无特异性临床表现,诊断依赖病理,治疗首选手术,预后尚存争议.本文报道郑州大学第一附属医院经治3例并复习相关文献以加深对其临床、治疗及预后特点的认知.

胃印戒细胞癌耐药基因与CEA、nm23表达的关系

目的 :探讨胃印戒细胞癌耐药基因特征与CEA、nm2 3表达的关系。方法 :对 5 3例胃印戒细胞癌的病理标本进行耐药基因人胎盘型谷胱甘肽S转移酶 (GST -π)、P170免疫组化检测 ,同时进行CEA、nm2 3免疫组化染色和超微结构观察。结果 :癌细胞呈现三种形态 :单核细胞样、浆细胞样及印戒细胞样。免疫组化检测观察 :5 3例胃印戒细胞癌组织CEA均呈阳性(10 0 %) ,nm2 3表达 43例 (81.13%) ,GST -π表达 38例 (71.6 9%) ,P170表达 11例 (2 0 .75 %) ;GST -π与CEA、nm2 3之间无显著性差异 (P >0 .0 5 ) ,与P170之间有显著性差异 (P <0 .0 5 )。结论 :GST -π在印戒细胞癌中的高表达与印戒细胞癌CEA分化具有密切的相关性 ,GST -π与P170负相关提示在印戒细胞癌中两种耐药基因表达具有不均一性 ,二者与nm2 3表达的关系不同 ,可能与CEA高度活性致组织转移抑制状况降低 ,加速癌细胞浸润转移 ,诱导产生耐药有关。