雌激素对雌激素受体阴性的子宫内膜腺癌细胞系JEC的作用

目的探讨不同浓度17β-雌二醇（E2）对雌激素受体（ER）阴性的子宫内膜腺癌细胞系JEC体外生长的影响。方法选用人子宫内膜腺癌细胞系JEC进行体外培养,加入含不同浓度E2的培养液,采用MTT比色、流式细胞术和激光共聚焦扫描显微镜的方法,观察子宫内膜腺癌细胞系JEC细胞的增殖活性、细胞周期时相变化及Ca2＋的变化。结果不同浓度的E2作用JEC细胞1、2、3 d后,其OD值和对照组比较无统计学差异,作用4、5 d后10-7、10-8和10-9mol/L组OD值和对照组比较差异有统计学意义（P〈0.05）,其中10-7mol/L组在第5天OD值与对照组比较有显著差异（P〈0.01）。10-7mol/L E2作用JEC细胞4、5 d后,G0/G1、S＋G2/M期细胞比例有所变化,但与对照组比较无统计学意义（P〉0.05）。10-7mol/L的E2作用JEC 4 d后Ca2＋的平均荧光强度值高于对照组（P〈0.01）。结论 ER阴性的JEC细胞生长可受到雌激素的调控,一定浓度的雌激素对ER阴性JEC细胞生长的调控可能与细胞内Ca2＋增加有关。     

NOD样受体家族蛋白3在中老年子宫内膜样腺癌组织中的表达及意义

目的探讨NOD样受体家族蛋白3（NOD-like receptor protein, NLRP3）在中老年患者子宫内膜样腺癌组织中的表达及临床意义。 方法选取2017年1月至2019年12月在浙江医院和浙江省肿瘤医院行手术切除的子宫内膜样腺癌患者35例,以及同期因子宫脱垂行全子宫切除术患者12例。采用实时荧光定量PCR、免疫印迹、免疫组化等方法检测比较不同子宫内膜组织中NLRP3 mRNA和蛋白的表达水平。同时应用子宫内膜样腺癌细胞系ishikawa和HEC-1A行体外细胞试验,观察NLRP3对子宫内膜样腺癌细胞增殖、侵袭及迁移能力的影响。两组间的比较采用t检验。 结果免疫组化检测：NLRP3在子宫内膜样腺癌组织中的表达水平明显高于正常子宫内膜组织,而且随FIGO分期及病理分级的增高而增高。实时荧光定量PCR检测：子宫内膜样腺癌组织中NLRP3 mRNA和NLRP3蛋白的表达水平均显著高于正常子宫内膜组织（t=3.769、4.316,P<0.05）。蛋白质印迹法：子宫内膜样腺癌组织中NLRP3蛋白表达明显强于正常子宫内膜组织。体外细胞试验：NLRP3 mRNA和NLRP3蛋白在ishikawa细胞和HEC-1A细胞中均有表达,而且两者在HEC-1A细胞中的表达水平均明显高于ishikawa细胞（t=2.373、2.559,P<0.05）;利用siRNA沉默子宫内膜样腺癌ishikawa细胞和HEC-1A细胞的NLRP3后,从第3天开始其增殖能力明显受限,并持续至细胞增殖实验结束的第7天;Transwell实验：沉默NLRP3后ishikawa细胞和HEC-1A细胞的迁移和侵袭数量均明显下降（t=7.343、6.571,3.859、3.289;P<0.05）。 结论NLRP3的表达与子宫内膜样腺癌细胞的增殖、侵袭和迁移能力密切相关。     

HER-2在子宫内膜样腺癌中的表达及临床意义

目的 探讨HER-2在子宫内膜样腺癌组织中的表达及意义.方法 采用免疫组化技术检测30例正常子宫内膜组织、21例子宫内膜不典型增生及51例子宫内膜样腺癌组织中HER-2的表达情况.结果 HER-2在正常子宫内膜组织、子宫内膜不典型增生及子宫内膜样腺癌组织阳性表达率分别为0、19.0％和31.4％;与正常子宫内膜组织相比,HER-2在子宫内膜不典型增生组织、子宫内膜样腺癌组织中的表达明显上调,差异有统计学意义（P均＜0.05）;HER-2的表达与子宫内膜样腺癌的病理分级、肌层浸润程度及淋巴结转移有关（P均＜0.05）.结论 HER-2的高表达可能与子宫内膜样腺癌的发生、发展密切相关.     

三氧化二砷对体外培养的人子宫内膜腺癌细胞凋亡的影响

目的研究三氧化二砷(As2O3)对体外培养的人子宫内膜腺癌JEC细胞株细胞凋亡的影响。方法采用体外细胞培养的方法,以透射电子显微镜观察人子宫内膜腺癌JEC细胞株形态变化,流式细胞仪分析细胞DNA水平及细胞周期,并测定细胞凋亡情况。结果(1)4.0μmol/LAs2O3作用72h,在电子显微镜下可见细胞核固缩、核碎裂、染色质边集等改变;8.0、16.0μmol/LAs2O3作用48h后,部分细胞出现染色质边集及核碎裂;(2)As2O3<2.0μmol/L时,G0/G1期和S期细胞明显减少,G2/M期细胞增多;As2O3为2.0、4.0μmol/L时,在细胞周期G1期前可见典型的细胞凋亡峰;As2O3为8.0～16.0μmol/L时,则主要表现为细胞坏死。结论As2O3可诱导人子宫内膜腺癌JEC细胞株发生细胞凋亡。     

子宫内膜腺癌组织中maspin、VEGF-C的表达变化及意义

目的观察子宫内膜腺癌组织中maspin和血管内皮生长因子C（VEGF—C）的表达变化,并探讨其意义。方法采用免疫组化sP法检测49例子宫内膜腺癌（腺癌组）、10例不典型增生（增生组）及15例正常子宫内膜（正常组）组织中的maspin蛋白和VEGF-C。结果腺癌组maspin、VEGF—C蛋白阳性率分别为57．1％、49．0％,增生组分别为40．0％、20．0％,正常组均为0。各组相比,P〈0．05。maspin表达与子宫内膜腺癌病理分期有关（P〈0．05）,VEGF—C表达与子宫内膜腺癌淋巴结转移、组织分级及肌层浸润有关（P均〈0．05）。子宫内膜腺癌组织中maspin、VEGF-C的表达呈正相关（r=0．519,P〈0．05）。结论子宫内膜腺癌组织中maspin、VEGF-C高表达,在子宫内膜腺癌的发生、浸润和转移中起重要作用。     

子宫内膜样腺癌中KLK7、E-Cad的表达及其与病情的相关性

目的探讨人角化层糜蛋白酶(KLK7)和E-钙黏蛋白(E-Cad)在子宫内膜样腺癌组织中的表达及其与病情的相关性。方法采用Western印迹检测10例增生期子宫内膜、10例不典型增生子宫内膜、30例子宫内膜样腺癌标本中KLK7、E-Cad蛋白的表达,分析其与各临床病理指标的相关性。结果 (1)KLK7、E-Cad在子宫内膜样腺癌中的蛋白表达与年龄无关;随着临床分期增高、组织分化降低、深肌层浸润、患者手术时有淋巴结转移,KLK7蛋白的阳性表达率呈上升趋势(P0.01),而E-Cad的阳性表达率呈下降趋势(P0.01,P0.05)。(2)在子宫内膜样腺癌中KLK7蛋白和E-Cad表达呈负相关(P0.05)。结论 KLK7蛋白在子宫内膜样腺癌中呈高表达,E-Cad在子宫内膜样腺癌中表达下降,两者可能与子宫内膜癌的病情进展及侵袭转移相关;KLK7蛋白和E-Cad在子宫内膜样腺癌组织中表达呈负相关,在子宫内膜癌侵袭转移中可能有协同效应。     

子宫内膜癌的病理类型及手术分期

1　病理类型子宫内膜癌最常见的病理类型为子宫内膜样腺癌 ,主要发生在围绝经期妇女 ,与高雌激素水平、子宫内膜增生有关 ,恶性度低 ,肌层浸润少 ,预后较好 ,占总子宫内膜癌的 80 %左右。其亚型有乳头型、分泌型、纤毛细胞型和有鳞状分化的腺癌。在子宫膜癌中 ,有 2 0 %左右的少见病理类型 ,其与雌激素水平、内膜增生无关 ,恶性度高 ,浸润性大 ,多发生于绝经后妇女 ,包括浆液性腺癌、透明细胞癌、鳞状细胞癌、未分化癌。其中浆液性乳头状癌 (U PSC)虽然仅占内膜癌的 5 %左右 ,但恶性度极高 ,早期即有深肌层、血管浸润及淋巴转移 ,极易发生…     

骨髓间充质干细胞对小鼠异位子宫内膜的作用观察

目的观察骨髓间充质干细胞能否迁移至异位子宫内膜并向子宫内膜细胞分化,探讨骨髓间充质干细胞与子宫内膜异位症的关系。方法构建子宫内膜异位症小鼠模型。分离雄性小鼠的骨髓间充质干细胞,用绿色荧光蛋白(GPF)标记后自鼠尾静脉注入模型体内。采用激光共聚焦显微镜观察模型小鼠异位子宫内膜组织中GPF的表达变化,用免疫荧光技术检测异位子宫内膜组织中GPF与角蛋白或波形蛋白复合表达变化。结果注射转染了GPF的骨髓间充质干细胞后第8天,异位子宫内膜碎片中有GPF表达,第20天表达最强,此后逐渐减弱;异位子宫内膜中,无GPF与角蛋白或波形蛋白阳性复合表达。结论骨髓间充质干细胞可迁移至异位子宫内膜组织中,但未发现其能分化为子宫内膜细胞。骨髓间充质干细胞与子宫内膜异位症的关系仍不确定。     

子宫内膜腺癌组织中E-钙黏素、TGF-β_1的表达变化及意义

目的观察子宫内膜腺癌组织中E-钙黏素、转化生长因子-β_1(TGF-β_1)的表达变化,并探讨其意义。方法采用免疫组化SP法检测29例正常增生期子宫内膜、25例不典型增生子宫内膜和41例子宫内膜腺癌组织中的E-钙黏素与TGF-β_1。结果正常增生期子宫内膜、不典型增生子宫内膜和子宫内膜腺癌组织中E-钙黏素的阳性表达例数分别为29、18、23例,TGF-β_1的阳性表达例数分别为11、12、28例,不同组织间两指标相比,P均<0.05。E-钙黏素的表达与子宫内膜腺癌病理分级、FIGO分期和淋巴结转移有关(P均<0.05),TGF-β_1的表达与子宫内膜腺癌肌层浸润深度、FIGO分期和淋巴结转移有关(P均<0.05);在子宫内膜腺癌原发灶中E-钙黏素与TGF-β_1的表达呈负相关(Kappa=0.31,P<0.05)。结论子宫内膜腺癌组织中E-钙黏素的表达下降、TGF-β_1的表达升高,二者的异常表达与子宫内膜腺癌恶性程度、转移和浸润有关。     

不同病理类型子宫内膜组织中β-catenin、YAP-1、Survivin 表达观察

目的观察不同病理类型子宫内膜组织中β-链蛋白(β-catenin)、Yes相关蛋白1(YAP-1)和生存素(Survivin)的表达变化,并探讨三者与子宫内膜样腺癌发病的关系。方法采用免疫组化法分别检测增殖期、单纯性增生、复杂性增生子宫内膜组织及子宫内膜样腺癌组织中的β-catenin、YAP-1、Survivin。结果β-catenin、YAP-1、Survivin在增殖期、单纯性增生、复杂性增生子宫内膜组织及子宫内膜样腺癌组织中的阳性表达依次增多(P均<0.05);β-catenin在高分化、中分化、低分化子宫内膜样腺癌组织中的阳性表达依次减少(P均<0.05)。子宫内膜样腺癌组织中β-catenin与YAP-1、YAP-1与Survivin阳性表达均呈正相关关系(r分别为0.378、0.700,P均<0.05)。结论子宫内膜样腺癌组织中β-catenin、YAP-1、Survivin阳性表达增高,三者可能协同参与了子宫内膜样腺癌的发病。     

奥曲肽对人子宫内膜癌HEC-1-B细胞增殖及其血管内皮生长因子表达的影响

目的探讨奥曲肽对人子宫内膜癌细胞增殖及其血管内皮生长因子（VEGF）表达的影响。方法加入不同浓度的奥曲肽对HEC-1-B细胞进行培养。细胞增殖用MTT法检测，细胞中VEGF的表达采用RT-PCR技术及酶联免疫吸附试验（ELISA）检测。结果加入奥曲肽培养后，HEC-1-B细胞的增殖受抑制．而且其VEGFmRNA指数降低，VEGF蛋白表达减少。结论奥曲肽能抑制人子宫内膜癌细胞增殖及VEGF的分泌。     

Identification of prognostic tumor-infiltrating immune cells in endometrial adenocarcinoma

To identify prognostic tumor-infiltrating immune cells of endometrial adenocarcinoma.The gene expression profiles of endometrial adenocarcinoma were downloaded from the Cancer Genome Atlas (TCGA). The abundance of tumor-infiltrating immune cells in endometrial adenocarcinoma samples was calculated by CIBERSORT algorithm. Kaplan–Meier analysis was used to identify prognostic tumor-infiltrating immune cells.This study identified 22 kinds of tumor-infiltrating immune cells. Macrophages M0 and CD8 T cells were prognostic factors of endometrial adenocarcinoma. The abundance of macrophages M0 (P = .038) was significantly correlated with better prognosis of endometrial adenocarcinoma. In contrast, the abundance of CD8 T cells (P = .049) was associated with poor prognosis of endometrial adenocarcinoma.Tumor-infiltrati macrophages M0 and CD8 T cells were prognostic factors of endometrial adenocarcinoma.     

Characterization of one newly established esophageal cancer cell line CSEC from a high-incidence area in China

SUMMARY.  The Chaoshan littoral is located in a high-incidence area of esophageal cancer in the south of China. In this study, a new esophageal cancer cell line CSEC was established from a 47-year-old female Chinese patient in this district. The biological characters of the cultured cells were investigated, including morphology, ultrastructure, growth kinetic features, tumorigenicity, expression of tumor-associated antigen and cytogenetic features. CSEC cell line grew continuously with a doubling time of 39.5 h and had been passaged over 80 times. The CSEC cells possessed features of squamous epithelial cells with cytokeratin indicated by immunohistochemical staining and tonofilaments and desmosomes revealed by electron microscopy. Tumorigenicity to severe combined immunodeficient mice was confirmed and the tumors developed revealed well-differentiated squamous cell carcinoma, similar to the origin tumor from which the cell line derived. The cytogenetic analysis demonstrated hypertetraploid karyotypes. Chromosome structure aberrations were common and complicated. Immunohistochemical staining showed that CSEC cells were infected with HPV and over-expressed p53. In summary, the CSEC cell line is a well-differentiated esophageal squamous cell carcinoma cell line from a high-incidence area in southern China. It may provide a useful model for the pathogenesis and therapeutic research of esophageal squamous cell carcinoma.     

Chemokine (C-C) motif ligand 20 is regulated by PGF2α-F-prostanoid receptor signalling in endometrial adenocarcinoma and promotes cell proliferation

Prostaglandin F_2α (PGF_2α) is an inflammatory mediator which signals through a G-protein coupled receptor, the F-prostanoid (FP) receptor. We have previously shown elevated FP receptor expression in endometrial adenocarcinoma, a common gynaecological malignancy in Western countries. In this study, the expression of the chemokine CC motif Ligand 20 (CCL20) was determined to be regulated by PGF_2α-FP receptor signalling in endometrial adenocarcinoma explants and cell line, and expression of CCL20 and its receptor CCR6 was elevated in endometrial adenocarcinoma compared to non-malignant endometrium. Both CCL20 and CCR6 were localised to neoplastic endometrial epithelial cells. The induction of CCL20 expression by PGF_2α-FP signalling in an endometrial adenocarcinoma cell line stably expressing the FP receptor (FPS cells) was found to be dependent on the intracellular signalling of Gq, EGFR, ERK, calcineurin and nuclear factor of activated T-cells (NFAT) proteins. The treatment of FPS cells with recombinant CCL20 caused a significant increase in proliferation. Therefore these data demonstrate a role for the FP receptor in regulation of the chemokine CCL20, which can mediate proliferation of endometrial adenocarcinoma epithelial cells.     

依西美坦对异位子宫内膜间质细胞caspase-3表达的影响和意义

目的探讨芳香化酶抑制剂依西美坦对异位子宫内膜间质细胞半胱氨酸蛋白酶-3(caspase-3)表达的影响及其临床意义。方法体外培养正常和异位子宫内膜间质细胞.应用RT-PCR检测芳香化酶mRNA的表达.用SABC免疫组织化学法检测caspase-3的表达,用流式细胞仪检测细胞凋亡情况。结果与正常内膜间质细胞相比,异位内膜间质细胞可以利用芳香化酶合成雌激素,caspase-3表达和细胞凋亡减少(P均<0.05);依西美坦灭活芳香化酶,降低雌激素水平,caspase-3表达和细胞凋亡升高(P均<0.05);反向添加雌激素可阻断依西美坦的作用。结论caspase-3调控异位内膜间质细胞的凋亡;依西美坦增加caspase-3的表达,促进异位内膜间质细胞凋亡。     

Alpha-fetoprotein-producing gastric carcinoma: Biological properties of a cultured cell line

We describe a gastric carcinoma cell line that has been maintained in vitro for more than 10 years and retains the capacity to produce a large amount of alpha-fetoprotein. This cell line was isolated from a metastatic lymph node of a 63-year-old male patient with advanced gastric carcinoma (T2N3P0H0M0) who showed high serum levels of alpha-fetoprotein. The primary tumor was moderately differentiated tubular adenocarcinoma and the lymph node was poorly differentiated adenocarcinoma without any particular pattern. The cultured cells grew as densely packed islet-like colonies with small polygonal cells. Electron microscopy revealed cells abundant in cytoplasmic organelles, with some cellular attachments being tight with junctional complexes and some being loose across intercellular spaces. The free cell surface had microvilli. The population doubling-time was 152 h at passage 58. Chromosomal analysis revealed the modal number to be 77, with numerous karyotype abnormalities. The tumorigenicity of the cultured cells in athymic nude mice was positive only when they were subcutaneously transplanted beneath a plastic plate, but when the cells were transplanted subcutaneously or administered by intrasplenic injection in intact or weakly irradiated nude mice, no tumorigenicty was shown. The cell line produced tumor-associated antigens, such as alpha-fetoprotein, carcinoembryonic antigen, and tissue polypeptide antigen. This cell line may be useful for comparative studies of different types of gastric carcinoma and alpha-fetoproteins of different origins.     

Establishment and characterization of an opisthorchiasis-associated cholangiocarcinoma cell line （KKU-100）

AM: To establish and characterize a new cholangiocarcinoma cell line from a patient living in the Opisthorchis viverrini (O. viverrini) endemic area of Northeast Thailand. METHODS: Fresh liver biopsy and bile specimens were obtained from a 65-year-old Thai woman with cholangiocarcinoma of the ports hepatis. After digestion, the cells were cultured in Ham's F12 media. The established cell line was then characterized for growth kinetics, cell morphology, imm-unocytochemistry and cytogenetics. Tumorigenicity of the cell line was determined by heterotransplanting in nude mice. RESULTS: The primary tumor was a poorly differentiated tubular adenocarcinoma. Examination of the bile revealed malignant cells with O. viverrin eggs. The cholangiocarcinoma cell line KKU-100 was established 4 mo after the primary culture-population doubling time was 72 h. KKU-100 possesses compact and polygonal-shaped epithelial cells. Immunocytochemically, this cell line exhibited cytokeratin, EMA, CEA, and CA125, but not a-fetoprotein (AFP), CA19-9, desmin, c-met, or p53. Such protein expressions parallel those of the primary tumor. Cytogenetic analysis identified aneuploidy karyotypes with a modal chromosome number of 78 and marked chromosomal structural changes. Inoculation of KKU-100 cells into nude mice produced a transplantable, poorly differentiated aden-ocarcinoma, similar to the original tumor. CONCLUSION: KKU-100 is the first egg-proven, Opisthorchis- associated cholangiocarcinoma cell line, which should prove useful for further investigations of the tumor biology of this cancer.     

Establishment and characterization of human hilar bile duct carcinoma cell line and cell strain

We established and characterized a human hilar bile duct carcinoma (HBDC) cell line from cells isolated from the ascites of a 75-year-old Japanese woman. Histopathological findings were confirmed to be poorly differentiated adenocarcinoma (pat BsrlCm according to the Japanese Society of Biliary Surgery General rules for surgical and pathological studies on cancer of the biliary tract, 4th edn.). Using a semi-agarose method, a daughter-cell strain was also established. Both the cell line and the strain were transplanted into scid or nude mice with a 100% inoculation rate. The population doubling times of the cell line and the strain were 32.25 and 35.78 h, respectively. The cell line and strain strongly expressed human epithelial antigen (HEA)-125 and cytokeratin (CK)-19 but did not express desmin and partly expressed vimentin. High values tumor markers (carbohydrate antigen [CA19-9], Span-1, KMO-1) were detected in culture supernatants from both the cell line and the cell strain and the concentrations paralleled the patient's serum data. DNA analysis revealed that the cell strain was diploid, whereas the cell line was aneuploid, with a DNA index (DI) of 0.85. Chromosomal analysis of the cell line and the strain revealed a range of numerical abnormalities (76–93 and 74–88, respectively) as well as structural abnormalities. The establishment of this HBDC cell line and strain may provide some benefit for fundamental biological research.