逆转录病毒介导p53基因对食管癌细胞株的生长抑制作用

目的观察人类野生型p53(wt p53)基因对人食管癌细胞系的抑制作用.方法用逆转录病毒为载体将外源性wt p53基因导入人食管癌细胞系ECA109,通过体外及小鼠体内实验研究转入基因的表达及对肿瘤的抑制作用.结果 p53在转染细胞ECA109/p53中表达水平提高.外源性wt p53基因的导入和表达能使ECA109细胞的生长速率减低,软琼脂集落形成能力下降,G0 G1期细胞比例增加、S期细胞比例降低,凋亡指数升高,裸鼠体内成瘤能力明显下降.结论逆转录病毒载体介导的外源性野生型p53能够抑制人食管癌细胞生长.     

野生型p53基因对人胆囊癌细胞生长及致瘤性的抑制作用

目的:研究野生型p53(wtp53)基因对人胆囊癌细胞生长及致瘤性的影响.方法:应用免疫细胞化学染色、PCR产物直接测序方法分析细胞系遗传背景,在脂质体介导下将含有wtp53的真核表达质粒pCMV-p53,导入GBC-SD细胞中.用G418筛选,建立转染克隆细胞系.以PCR,RT-PCR和蛋白印迹证实外源p53基因的整合与表达;以细胞生长曲线和集落形成实验反映细胞增殖状况;以裸鼠移植瘤试验检测体内致瘤性的影响.结果:GBC-SD细胞P53蛋白过表达;直接测序发现第5外显子126位密码子存在TAC→AAC的碱基突变.外源p53基因已整合入转染后的GBC-SD细胞并获稳定表达.表达外源wtp53的GBC-SD-wtp53细胞生长速率减慢、集落形成能力下降及裸鼠致瘤性受到显著抑制.结论:野生型p53基因可有效抑制人胆囊癌GBC-SD细胞的体内、外生长.     

食管癌预后相关肿瘤标志物

与食管癌预后相关的肿瘤标志物可分为蛋白质类和糖蛋白类,前者包括细胞角蛋白19片段抗原21-1(CYFRA21-1)、p53抗体等,后者包括鳞状细胞癌抗原、血管内皮生长因子(VEGF)等,对其检测有利于预测食管癌的预后.     

食管癌预后相关肿瘤标志物

与食管癌预后相关的肿瘤标志物可分为蛋白质类和糖蛋白类,前者包括细胞角蛋白19片段抗原21-1(CYFRA21-1)、p53抗体等,后者包括鳞状细胞癌抗原、血管内皮生长因子(VEGF)等,对其检测有利于预测食管癌的预后.     

食管癌预后相关肿瘤标志物

与食管癌预后相关的肿瘤标志物可分为蛋白质类和糖蛋白类,前者包括细胞角蛋白19片段抗原21-1(CYFRA21-1)、p53抗体等,后者包括鳞状细胞癌抗原、血管内皮生长因子(VEGF)等,对其检测有利于预测食管癌的预后.     

食管癌预后相关肿瘤标志物

与食管癌预后相关的肿瘤标志物可分为蛋白质类和糖蛋白类,前者包括细胞角蛋白19片段抗原21-1(CYFRA21-1)、p53抗体等,后者包括鳞状细胞癌抗原、血管内皮生长因子(VEGF)等,对其检测有利于预测食管癌的预后.     

食管癌预后相关肿瘤标志物

与食管癌预后相关的肿瘤标志物可分为蛋白质类和糖蛋白类,前者包括细胞角蛋白19片段抗原21-1(CYFRA21-1)、p53抗体等,后者包括鳞状细胞癌抗原、血管内皮生长因子(VEGF)等,对其检测有利于预测食管癌的预后.     

食管癌预后相关肿瘤标志物

与食管癌预后相关的肿瘤标志物可分为蛋白质类和糖蛋白类,前者包括细胞角蛋白19片段抗原21-1(CYFRA21-1)、p53抗体等,后者包括鳞状细胞癌抗原、血管内皮生长因子(VEGF)等,对其检测有利于预测食管癌的预后.     

食管癌预后相关肿瘤标志物

与食管癌预后相关的肿瘤标志物可分为蛋白质类和糖蛋白类,前者包括细胞角蛋白19片段抗原21-1(CYFRA21-1)、p53抗体等,后者包括鳞状细胞癌抗原、血管内皮生长因子(VEGF)等,对其检测有利于预测食管癌的预后.     

食管癌预后相关肿瘤标志物

与食管癌预后相关的肿瘤标志物可分为蛋白质类和糖蛋白类,前者包括细胞角蛋白19片段抗原21-1(CYFRA21-1)、p53抗体等,后者包括鳞状细胞癌抗原、血管内皮生长因子(VEGF)等,对其检测有利于预测食管癌的预后.     

Partial suppression of tumorigenicity in a human lung cancer cell line transfected with Krev-1.

A human non-small-cell lung carcinoma cell line, Calu-6 (from an anaplastic carcinoma), was transfected with the Ki-ras-related anti-oncogene Krev-1. Several transfectant lines were obtained that showed a reduced tumorigenicity in nude mice with respect to the parental and control transfected cell lines. This decrease was approximately 50% in tumor incidence at 4 wk after subcutaneous inoculation of the transfected cells. In addition, the volume of the Calu-6 revertant-derived tumors was three to 10 times smaller than that of the equivalent tumors produced by inoculation of the control cell line transfected with the neomycin-resistance gene. Krev-1--transfected cells that exhibited reduced tumorigenicity expressed Krev-1 mRNA and had variable numbers of copies of the Krev-1 gene. Moreover, Krev-1--transfected cells exhibited a more differentiated squamous epithelial morphology than the parental and control cell lines did. Moderately elevated levels of protein kinase C activity were detected in some revertant clones. Such activity correlated with the level of expression of Krev-1 mRNA in most cases. In summary, Krev-1 induced important morphological and biological changes in transfected Calu-6 cells that we interpreted as partial reversion of the malignant phenotype.     

食管梭形细胞癌临床病理分析

目的分析食管梭形细胞癌的临床病理特点,探讨其组织来源。方法回顾性分析4例食管梭形细胞癌的临床病理资料及复习有关文献。结果 4例患者为男性,1例为女性,平均年龄57.2岁,病变位于食管中下段,大体呈息肉型、髓质型、蕈伞型及溃疡型。显微镜下可见鳞状细胞癌和肉瘤样成份,其中3例未找到两者相移行的区域。免疫组织化学检查发现肉瘤样区域细胞中有AE1/AE3、vimentin蛋白表达,息肉型者软骨肉瘤样区域仅有Vimentin和S-100蛋白表达,纤维肉瘤成份仅有vimentin蛋白表达。结论食管梭形细胞癌本质上是1种特殊类型的癌,包括肉瘤样癌和癌肉瘤。肉瘤样癌区域可能是癌细胞双向分化的结果,癌肉瘤的发生机制有待于进一步研究。     

食管癌细胞株高侵袭力亚系的建立及其生物学特性研究

目的:建立具有不同转移潜能的高侵袭能力食管癌细胞株亚系并研究其生物特性.方法:利用Transwell侵袭小室从人食管鳞癌Eca-109细胞株筛选高侵袭能力食管癌亚系,HE染色比较细胞形态;四甲基偶氯唑蓝(MTT)法检测细胞增殖能力的变化;流式细胞术( FCM)检测细胞周期;蛋白质印迹法检测与侵袭能力相关基质金属蛋白酶-2(MMP-2)和金属蛋白醇组织抑制因子-2(TIMP-2)蛋白表达.结果:利用Transwell侵袭小室从食管癌Eca-109细胞株中筛选出高侵袭能力食管癌细胞株亚系,命名为Eca-109 T4.2个细胞系细胞形态没有明显差异.MTT法检测显示,亚系Eca-109 T4增殖能力强.细胞周期显示,增殖指数(PI)高,PI=41.0％,且MMP-2蛋白表达相比增高,P=0.023;TIMP-2蛋白表达相比增高,但差异无统计学意义,P=0.392.结论:建立了不同转移潜能的高侵袭能力食管癌细胞株亚系,将可以用于探讨食管癌转移机制的研究.     

高迁移率族蛋白HMGB1在宫颈鳞癌组织中的表达

目的:探讨宫颈鳞癌组织中高迁移率族蛋白HMGB1表达情况及其与肿瘤分化程度、大小、浸润和转移的关系。方法:应用组织芯片免疫组织化学方法检测了30例宫颈原位癌,90例无转移的宫颈癌,30例有转移的宫颈癌和30例正常宫颈鳞状上皮中HMGB1表达情况,并应用western blot技术定量检测其表达情况。结果:HMGB1在宫颈鳞癌组织中呈强阳性表达58.7%(88/150),正常宫颈鳞状上皮中弱表达,差异具有显著性(P<0.01),HMGB1阳性率与肿瘤的大小、浸润及转移呈正相关(P<0.01);与肿瘤分化程度无关(P>0.05)。结论:HMGB1在宫颈鳞癌组织中尤其是发生转移的宫颈鳞癌组织中呈强阳性表达,可作为宫颈鳞癌浸润、转移及预后的重要判定指标之一,并有望成为基因治疗的靶点。     

Establishment and analysis of biological characteristics of a human duodenal carcinoma cell line, WDC-1

BACKGROUND: Duodenal carcinoma is very rare and its culture cell lines have rarely been established. METHODS: Tumor cells separated from a surgically resected primary tumor of duodenal carcinoma were put into culture. The patient was an 81-year-old female and had metastatic lymph nodes. We investigated the biological characteristics of the culture cells including in vitro cell kinetics, karyotype, expression of tumor markers and integrins and tumorigenicity and histology in nude mice. RESULTS: A new cell line, designated WDC-1, was established. This duodenal carcinoma cell line proliferated in a monolayered sheet with a doubling time of 50 h. The histological findings of the xenograft in nude mice were similar to those of the primary tumor. WDC-1 cells produced carcinoembryonic antigen and expressed 1 integrin and very late antigen (VLA)-4d in vitro. CONCLUSIONS: A duodenal carcinoma cell line was established, which is rare and may contribute to progress in understanding the biological features of duodenal cancer.     

小鼠子宫颈癌U14细胞系的建立及生物学特性研究

目的:建立小鼠宫颈癌肿瘤细胞系,观察其生物学特性。方法:采用肿瘤原代培养法对小鼠U14肿瘤进行体外传代培养,定期对培养细胞进行形态学观察,进行免疫组织化学鉴定、细胞周期检查、染色体检测、细胞生长测定,进行615、C57BL/c小鼠移植观察体内成瘤情况,并用pEGFP-N1质粒转染U14细胞。结果:细胞呈贴壁和悬浮混合生长,CK阳性特征。体外连续培养10个月,传代50代,细胞倍增时间为21.83小时,细胞周期测定G1期为34%,G2期为26.4%,S期为39.6%。培养U14细胞3-4天处于对数生长期,染色体为亚四倍体核型,众数为64～84条。615小鼠、C57BL/c小鼠移植成瘤率均为100%,无支原体污染。建立了GFP( )的U14-GFP单克隆细胞株。结论:成功建立了小鼠U14子宫颈癌细胞系及GFP标记的U14-GFP细胞株,体内移植可建立常规及带荧光标记的肿瘤模型,可用于体内体外相结合的肿瘤研究。     

Clinical pathology analysis of esophageal sarcomatoid carcinoma

Objective: The purpose of this study was to study the clinical, imaging characters and pathological characteristics of esophageal sarcomatoid carcinoma. Methods: We reviewed 23 cases of esophageal sarcomatoid carcinoma from Janu-ary 2006 to December 2013 in four hospitals. The data of patients who were esophageal sarcomatoid carcinoma operated were retrospectively analyzed. Al cases had completed upper gastrointestinal barium images materials and 14 of these cases had completed CT images materials. Upper gastrointestinal barium images and CT imaging features include tumor location, size, shape, and strengthen, etc. The biological parameters of lesions including the express of cytokeratin AE1/AE3, 34βE12, p63, Vimentin, desmin, Actin, S-100 and Ki-67 detected by immunhistochemical UltraSensitiveTM S-P method (n=23), and the patients’ data of contrastographic picture (n=23), imaging characters of CT scan (n=14), and their relationship were studied. Results:Upper gastrointestinal barium images, CT imaging and gastrointestinal fiberscopy revealed lobulated intraluminal fil ing defect 0.4 cm to 5.7 cm × 3.5 cm × 1.3 cm (mean=3. 7 cm) in the mid (n=14), lower (n=7) and upper (n=2) intrathoracic esophagus. Among 23 cases of esophageal sarcomatoid carcinoma, 19 patients were of mushroom type, 2 patients was of ulcer type, and 2 patients were of medul a type;19 patients were pedunculated, and 4 patients were no pedunculated (2 patients was of ulcer type). The tumor surface was relatively smooth and esophageal compliance was maintained. The pathological changes of esophagus such as lightly locked, rigid wal no-manifest partly, esophageal lumens expand partly, major fil ing sublobe defect could be shown through contrast medium. Normal esophagus was no unpack obviously over pathological changes. Enhanced computed tomography showed tumors in the intrathoracic esophagus and 8 lymph nodes metastases in 3 cases. Histological y, carcinomatous and sarcomatous components coexist. Microscopical y, the tumor comprised poorly dif erentiated squamous cellcarcinoma and spindle-shaped cells resembling leiomyosarcoma. Immu-nohistochemical y, spindle-shaped sarcomatous cells displayed weekly positive reaction to cytokeratin AE1/AE3. Transitional zone was seen between sarcomatous and carcinomatous elements in 5 cases. The 17 lymph nodes metastases in 5 cases (53 lymph nodes) among 23 cases esophageal sarcomatoid carcinoma (187 lymph nodes) were observed. Conclusion:The clinical and radiologic features of esophageal sarcomatoid carcinoma overlap with those of other esophageal neoplasms. There are the radiologic imaging changes such as a large, intraluminal, polypoid mass, major fil ing sublobe defect and pedicle skin flap tumor in esophageal lumen, esophageal lumen extension partly, dissepiment rigidity wal no obviously, etc. Histological y, carcinomatous and sarcomatous components coexist and the biphasic pattern is the key diagnostic feature. However, esophageal sarcomatoid carcinoma has a more favorable prognosis than other malignant esophageal neoplasms. Immunohistochemical staining seems necessary to distinguish these lesions from other esophageal neoplasms.     

腺病毒介导的p53基因对喉癌细胞生长的抑制作用

目的探索ｐ５３基因在喉癌基因治疗方面的可行性。方法以人喉癌细胞系Ｈｅｐ－２为实验对象,将载有人野生型ｐ５３ｃＤＮＡ并含巨细胞病毒（ＣＭＶ）启动子的重组腺病毒（Ａｄ５ＣＭＶ－ｐ５３）感染Ｈｅｐ－２细胞及肿瘤组织,体内外实验观察Ａｄ５ＣＭＶ－ｐ５３对Ｈｅｐ－２细胞生长的影响。结果当Ａｄ５ＣＭＶ－ｐ５３在１００ＭＯＩ效靶比时,全部Ｈｅｐ－２细胞得到转染。感染２天后ｐ５３蛋白表达达到高峰,Ｈｅｐ－２生长受到明显的抑制。Ａｄ５ＣＭＶ－ｐ５３感染Ｈｅｐ－２细胞在裸鼠中失去致瘤性。瘤内注射Ａｄ５ＣＭＶ－ｐ５３后,荷瘤裸鼠的肿瘤体积明显减小。结论Ａｄ５ＣＭＶ－ｐ５３转导野生型ｐ５３基因可能是一种有效的喉癌基因治疗途径。     

New Human Oral Squamous Carcinoma Cell Line and Its Tumorigenic Subline Producing Granulocyte Colony-stimulating Factor

A new human carcinoma cell line, MISK81-5, was established from a metastatic lymph node of oral squamous cell carcinoma. Immunocytochemical and ultrastructural observations revealed an obvious epithelial origin of the cell line. Chromosome analysis revealed a hypertriploid karyotype with numerical and structural anomalies. MISK81-5 cells could form a tumor mass in the subcutaneous tissue of recipient BALB/c athymic mice only when coinjected with Matrigel. A stem cell assay revealed that conditioned medium (CM) of MISK81-5 contained granulocyte colony-stimulating factor (G-CSF) or interleukin-6 activity. Quantitation by ELISA disclosed a higher concentration of G-CSF in the CM of MISK81-5 than in the CM of other squamous and gastric carcinoma cell lines. The sMISK, that was derived from MISK81-5 as a subpopulation of the cell line having higher tumorigenicity, also showed a similar hematopoietic stimulating activity to that of MISK81-5. These characteristics of the MISK81-5 cell line and its subpopulation, sMISK will be useful for studying the biological behavior of oral squamous cell carcinomas and its relation to hematopoietic stimulating factors.     

高表达MUC1的人食管癌裸鼠移植瘤模型的建立

背景与目的:Mucin-1(MUC1)粘蛋白是一种肿瘤相关抗原,是肿瘤免疫治疗良好的分子靶点之一.本研究建立高表达MUCl的人食管癌裸鼠皮下移植瘤模型,为研究以MUC1为靶点的食管癌的生物治疗提供体内模型.方法:以体外培养的高表达MUC1的人食管癌细胞株EC-109接种于4～5周龄的BALB/c裸小鼠皮下,观察移植瘤生长情况,对移植瘤进行病理组织学检查,采用免疫组化方法检测移植瘤细胞增殖细胞核抗原(proliferating cell nuclear antigens,PCNA),采用流式细胞仪检测移植瘤细胞的细胞周期及MUC1的表达.结果:裸鼠皮下移植瘤成瘤率为86.0%,移植瘤具有与人恶性肿瘤相似的组织学和生物学特点,其平均PCNA标记指数为(63.5 3.6)%、S期细胞百分比(S-phase fraction,SPF)平均值为(37.6±3.7)%、MUC1的平均表达率为97.5%.结论:高表达MUC1的人食管癌裸鼠皮下移植瘤模型具有恶性肿瘤的生物学特性,可用于研究人食管癌的生物学特点,同时为以MUC1为靶点的食管癌的免疫治疗研究提供了良好的体内模型.