乙酰胆碱与去甲肾上腺素对食管癌细胞分化调节作用及其机制的研究

目的：探讨食管神经递质对食管癌细胞分化的影响及其机制，为防治食管癌与复发提供实验依据。方法：应用EC109细胞系，分为乙酰胆碱（acetylcholine，Ach）组、Ach加阿托品组、去甲肾上腺素（norepinephrine，NE）组、NE加托拉唑林组、Ach＋NE组和对照组，体外培养14d，HE和氧化还原修复酶-1（peroxiredoxin-1，PRX1）免疫组化染色，镜下观察和图像分析，并对结果进行统计学处理。结果：各药物处理组与对照组相比：1）细胞3d长出突起，7d明显长长，14d部分细胞之间相互连结形成网状；2）PRX1免疫组化显色明显加深；3）核质比下降。Ach和NE组3项指标与对照组者相比差异均有统计学意义，P值均〈0．05。结论：神经递质Ach和NE对食管癌细胞分化具有诱导作用，其作用可能与其增强DNA修复酶的表达有关，但其确切作用机制尚待进一步研究。该发现为开拓食管癌发病机制和防治研究的新领域提供了基础资料。     

去甲斑蝥素诱导人食管癌Eca-109细胞凋亡及其作用机制

目的:探讨去甲斑蝥素诱导人食管癌Eca-109细胞的凋亡及其可能的作用机制.方法:去甲斑蝥素作用食管癌Eca-109细胞后,应用MTT法检测其对细胞的生长抑制作用, 透射电子显微镜下观察细胞超微结构的变化,琼脂糖凝胶电泳观察其对细胞凋亡的诱导作用,RT-PCR法检测caspase 8和caspase 3 mRNA的表达,Western 印迹法检测Fas、细胞型含死亡域的Fas结合蛋白样白介素-1β转换酶抑制蛋白(cellular FADD-like interleukin-1β converting enzyme inhibitory protein,c-FLIP) 、caspase 8和caspase 3蛋白的表达.结果:去甲斑蝥素对人食管癌Eca-109细胞具有生长抑制作用,并呈时效和量效依赖关系.电子显微镜下观察发现, 食管癌Eca-109细胞趋于凋亡.DNA琼脂糖凝胶电泳可见典型的DNA梯状条带.RT-PCR检测显示,caspase 8和caspase 3 mRNA的表达水平明显上升 (P<0.01).Western 印迹法检测结果显示,与阴性对照组比较,去甲斑蝥素作用后,食管癌Eca-109细胞中Fas、caspase 3和caspase 8蛋白的表达明显上升(P<0.05),c-FLIP蛋白的表达水平明显下降(P<0.05).结论:去甲斑蝥素能够抑制食管癌Eca-109细胞的生长并诱导其凋亡,其机制可能是通过上调Fas、caspase 8、caspase 3的表达和下调c-FLIP的表达来实现的.     

去甲斑蝥素诱导人食管癌Ec9706细胞凋亡及其可能的机制

目的：研究去甲斑蝥素对人食管鳞癌Ec9706细胞的致凋亡作用及其可能的作用机制,为去甲斑蝥素应用于临床抗癌治疗提供实验依据。 方法：不同质量浓度去甲斑蝥素 (0、5、10、20、40 μg/ml)分别作用Ec9706细胞不同时间(12、24和48 h)后,MTT方法检测细胞增殖抑制率,流式细胞术检测细胞凋亡及Caspase-3和Survivin蛋白表达的变化。结果：去甲斑蝥素作用后Ec9706细胞呈现不同程度的增殖抑制,而且细胞增殖抑制程度随作用剂量及时间增加不断增强,40 μg/ml去甲斑蝥素作用48 h时,Ec9706细胞增殖抑制率达（80.00±2.15）%。去甲斑蝥素显著诱导Ec9706细胞凋亡,其20 μg/ml作用48 h时,Ec9706细胞的凋亡率达（38.57±1.76）%。去甲斑蝥素作用后,Ec9706细胞中Caspase-3蛋白水平显著增高,而Survivin蛋白水平显著降低(P<0.05)。结论：去甲斑蝥素明显诱导食管癌Ec9706细胞凋亡,其作用机制可能与下调细胞Survivin蛋白及上调Caspase-3蛋白表达有关。     

去甲斑蝥素诱导人食管癌Ec9706细胞凋亡作用及机制研究

目的 研究去甲斑蝥素作用人食管鳞癌细胞Ec9706后细胞凋亡率,并进一步探讨去甲斑蝥素诱导Ec9706细胞凋亡的作用机制,为去甲斑蝥素应用于临床抗癌药物提供基础实验。 方法 不同浓度去甲斑蝥素 (0、5、10、20、40 μg/ml)分别作用Ec9706细胞不同时间(12、24和48h)后MTT方法检测细胞生长抑制率。不同浓度去甲斑蝥素(0、5、10、20 μg/ml) 分别作用Ec9706细胞不同时间(24和48h)后流式细胞术检测细胞凋亡及Caspase-3和Survivin蛋白的表达。结果 MTT结果显示,去甲斑蝥素作用Ec9706细胞后呈现不同程度的细胞生长抑制作用,而且随作用浓度及时间增加细胞生长抑制率呈现显著增高趋势。去甲斑蝥素作用Ec9706细胞后,流式细胞术检测结果显示,Ec9706细胞凋亡率与对照组相比显著增高(P<0.05),Caspase-3蛋白表达量显著增高而Survivin蛋白表达水平显著降低(P<0.05)。结论 去甲斑蝥素具有抑制食管癌Ec9706细胞生长的作用,其作用机制可能与下调细胞Survivin蛋白及上调Caspase-3蛋白表达水平,从而诱导食管癌细胞凋亡有关。     

环氧化酶-2与肝肿瘤

环氧化酶-2(COX-2)是环氧化酶的诱导型,是催化花生四烯酸合成前列腺素的限速酶.COX-2过表达与肝肿瘤有密切关系,其作用机制可能涉及多个方面.选择性COX-2抑制剂的深入研究有望使其在肝脏肿瘤的治疗方面有新的突破.     

去甲肾上腺素水平在胸腔积液鉴别诊断及预后评估中的价值

目的：探究胸腔积液中能够提示患者临床诊断与预后的潜在生化指标,为临床诊断与治疗提供帮助。方法：收集郑州大学第一附属医院2020年4月至2021年5月41例患者的胸腔积液样本,采用流式细胞术检测肿瘤相关巨噬细胞（tumor-associated macrophages,TAMs）比例,酶联免疫吸附测定（enzyme linked immunosorbent assay,ELISA）检测乳酸、去甲肾上腺素（norepinephrine,NE）水平,同时收集临床检测乳酸代谢相关分子乳酸脱氢酶（lactate dehydrogenase,LDH）结果,结合患者临床诊断与预后情况,通过独立样本t检验、逻辑回归分析、ROC分析等方法分析其与良恶性胸腔积液鉴别诊断及患者预后之间的关联。结果：恶性胸腔积液（malignant pleural effusion,MPE）中NE的水平明显高于非恶性胸腔积液（non-malignant pleural effusion,NMPE）[(5.24±3.49)ng/mL vs.(2.56±2.25)ng/mL,P<0.05];且NE水平与恶性病理诊断呈正相关...     

乙酰胆碱及其受体的非神经递质作用和肿瘤

乙酰胆碱（acetylcholine，ACh）是一种经典的兴奋性神经递质，通过结合特异受体，在神经细胞之间或神经细胞与效应器细胞之间中起着信息传递作用。ACh及其受体存在于从细菌到人类、从神经细胞到其他多种非神经细胞中，提示它是一类与系统发生相关的古老分子，可能不仅仅具有作为生理性递质的传递功能。多种人类疾病与ACh及其受体相关，尤其是近年来的研究发现ACh及其受体在多种肿瘤中发挥自分泌和旁分泌因子作用，参与细胞的生长调节，甚至与肿瘤的发生发展相关。因此，ACh涉及到神经系统外非胆碱能传递的作用显得格外引人注目，可能成为新的肿瘤治疗靶标。     

食管鳞癌COX-2蛋白表达意义研究进展

与对食管腺癌的作用相似,环氧合酶-2(COX-2)的表达也可能影响食管鳞癌的发生、发展及预后.多数食管鳞癌有COX-2蛋白不同程度的表达,但其真正临床意义尚未明了.COX-2选择性抑制剂可抑制食管鳞癌细胞生长,诱导细胞凋亡,可能对化疗和放疗起增敏作用.     

Effects of dietary zinc deficiency on esophageal squamous cell proliferation and the mechanisms involved

BACKGROUNDDietary zinc deficiency has been shown to be associated with the development of esophageal cancer in humans, but the exact mechanism of action is not knownAIMTo observe the effects of dietary zinc deficiency on esophageal squamous cell proliferation. METHODSThirty C57BL/6 mice were randomly divided into three groups: A zinc-sufficient (ZS) group, zinc-deficient (ZD) group, and zinc-replenished (ZR) group. For weeks 1–10, zinc levels in the mice diets were 30.66–30.89 mg/kg in the ZS group and 0.66–0.89 mg/kg in the ZD and ZR groups. During weeks 10–12, the ZR group was switched to the ZS diet; the other two groups had no changes in their diets. Changes in body weight, serum, and esophageal tissue zinc concentrations were assessed as well as differences in the expression of proliferating cell nuclear antigen (PCNA), mitogen-activated protein kinase p38 (p38MAPK), nuclear factor kappa B (NF-κB) p105, NF-κB p65, and cyclooxygenase (COX)-2 proteins in the esophageal mucosa.RESULTSThe body weight and zinc concentration in the serum and esophageal mucosa were significantly lower in the ZD and ZR groups than in the ZS group (P < 0.05). In ZD mice, there was a marked proliferation of basal cells in the esophageal mucosa, resulting in a disturbance in the arrangement of basal cells in layers 2–4, a thickening of the squamous layer, and a significant increase in the expression of the above-mentioned five proteins involved in proliferation and inflammation in the esophageal mucosa. Two weeks after switching to the ZS diet, the serum zinc concentration in the ZR group increased, and the expression of PCNA, NF-κB p105, and COX-2 decreased, but the concentration of zinc in the esophageal mucosa and the structure of the esophageal mucosa did not display any significant changesCONCLUSIONThe ZD diet decreased the growth rate and promoted the proliferation of esophageal squamous cells in mice. The mechanism of proliferation was related to the induced overexpression of COX-2, P38, PCNA, and NF-κB (p105 and p65), and the ZR diet reduced the expression of PCNA, NF-κB p105, and COX-2, thereby reversing this process.​     

Growth and differentiation of human esophageal carcinoma cell lines

Human esophageal carcinoma cell lines (8 cell lines) differed from their normal counterpart in terms of their morphological appearance, growth properties, and the expression of certain differentiated functions, namely keratin proteins and cross-linked envelopes. In contrast to normal human esophageal keratinocytes, the carcinoma cells were pleomorphic and tended to pile up in an unorganized fashion. When grown under optimal growth conditions the carcinoma cells generally grew to a higher saturation density than their nontransformed counterpart; their generation times were variable. Transformed cells grew better under stringent growth conditions (e.g., decreased serum and no additional growth factors except hydrocortisone) than did nontransformed human esophageal keratinocytes but their growth was still restricted under these conditions. The carcinoma cells retained a requirement for a 3T3 feeder layer when grown at clonal densities (5 X 10(3) cells/60-mm dish) but could be passaged and maintained without a feeder layer if plated at higher than clonal densities (10(5) cells/60-mm dish). All cell lines grew in an anchorage-independent fashion in soft agarose although the colony forming efficiency and size of the colonies varied among the different cell lines. Not all anchorage-independent cell lines were tumorigenic. Tumorigenic potential was greatly augmented by the use of cell lines derived from soft agarose selected clones. Altered expression of keratin proteins and cross-linked envelopes was observed in the carcinoma cell lines and generally reflected those changes seen in primary esophageal carcinomas. In two cell lines (HCE-4 and HCE-6), the synthesis of the Mr 44,000 (analogous to Rheinwald's Mr 40,000 keratin) and 52,000 keratins was suppressed coincident with the appearance of the 67 Kd keratin in tumors derived from these cell lines. These keratin patterns were once again reversed in cell lines recultured from these tumors, suggesting that the expression of these specific keratins is subject to extrinsic growth regulation. Another feature of terminal differentiation in keratinocytes, cross-linked envelope formation, was found to be significantly altered (reduced) in most but not all human esophageal carcinoma cell lines.     

食管鳞状细胞癌的肿瘤血管生成机制

血管内皮生长因子、缺氧诱导因子、白细胞介素、血管生成素样蛋白、整合素和上皮间质转化在食管鳞状细胞癌血管生成中为癌细胞的生长、侵袭和转移提供了营养支持和有利环境.对食管鳞状细胞癌血管生成机制的研究将为其抗血管靶向治疗提供更多思路和潜在靶点.     

靛玉红甲肟对人食管癌细胞株EC-1和Kyse70体外增殖和细胞周期的影响

目的:探讨靛玉红甲肟(indirubin-3'-monoxime)对体外培养的人食管癌细胞株EC-1和Kyse70细胞增殖和细胞周期的影响.方法:不同浓度靛玉红甲肟处理食管癌EC-1和Kyse70细胞后,采用MTT法检测细胞的增殖活性,FCM法和RT-PCR法检测细胞周期分布及bcl-2和bax mRNA表达的变化.结果:靛玉红甲肟对人食管癌细胞EC-1和Kyse70的生长具有明显的抑制作用,且表现为剂量依赖性和时间依赖性(P<0.05).FCM法检测发现,以5 μmol/L的靛玉红甲肟对EC-1细胞处理24、48和72 h后,G0/G1期细胞比例逐渐下降(P<0.05),S期细胞比例未见明显改变,G2/M期细胞比例逐渐上升(P<0.05),呈时间依赖性.RT-PCR法检测发现人食管癌细胞bcl-2和bax mRNA比例呈时间依赖性下调.结论:靛玉红甲肟对人食管癌细胞EC-1和Kyse70具有明显的增殖抑制作用,其作用机制可能与阻滞细胞周期G2/M期和下调bcl-2/bax 表达比例有关     

Cytogenetic studies on 3 esophageal cancer cell lines

The cytogenetic studies on three esophageal cancer cell lines established in China were done. Thirty metaphases showing suitable chromosome length and good G-banding pattern from each of the cell lines were chosen and subjected to karyological analysis. The typical karyotypes from these cell lines were listed, and the variation of chromosome number as well as the morphological characteristics, possible sources and the incidence of the marker chromosomes were analysed. Eca 109 is the cell line first established and used extensively in China. A strictly regular karyotype pattern was found in it: the modal number of chromosomes being 63-64; the number of each type of chromosome varying between 1-5; a distal deletion of short arm of chromosome No. 1 being discernible in all metaphases, with break sites located within 1p22-1p33. Also a distal deletion of long arm of chromosome No. 4 was usually visible. There were seven marker chromosomes with high incidence. Among them, M1 marker chromosome was a large subacrocentric chromosome which was observed in the early passages of this cell line. The chromosome number of Ec 17 cell line was usually subtetraploid. In addition to the numerical variation in some of the chromosomes, six marker chromosomes were usually observed. Among them, M1 involved reciprocal translocation between chromosome No. 1 and No. 4. M3 of Ec 17 was in correspondence with M5 of Eca 109. Both were Rob (13q;14q). The chromosome number of Ec 56 was usually subtetraploid, and in addition to the numerical variation in some of the chromosomes, seven marker chromosomes were usually observed.     

Flavonoids and risk of squamous cell esophageal cancer

The relation between 5 classes of flavonoids (flavanones, flavan-3-ols, flavonols, flavones and anthocyanidines) and esophageal cancer was investigated using data from a case-control study conducted between 1992 and 1997 in 3 areas of northern Italy. The study included 304 cases (275 men, 29 women) with a first diagnosis of squamous-cell carcinoma of the esophagus and 743 controls (593 men, 150 women) with no history of cancer, admitted for acute illnesses, unrelated to tobacco and alcohol consumption, to major hospitals of the areas under surveillance. Dietary habits were investigated using a validated food frequency questionnaire. Odds ratios (ORs) and 95% confidence intervals (CI) were computed after allowance for age, sex, study centre, years of education, alcohol drinking, tobacco smoking, body mass index and energy intake. An inverse association emerged between flavanone intake and esophageal cancer risk (OR=0.38 for the highest vs. the lowest quintile, 95% CI=0.23-0.66). The inverse relation between flavanones and esophageal cancer tended to be stronger in those who drank >or=6 drinks/day. In conclusion, this study suggests that flavanone intake is inversely associated with esophageal cancer risk and may account, with vitamin C, for the protective effect of fruit, especially citrus fruit, on esophageal cancer.     

RANKL-dependent and RANKL-independent mechanisms of macrophage-osteoclast differentiation in breast cancer

The cellular and humoral mechanisms accounting for tumour osteolysis in metastatic breast cancer are uncertain. Osteoclasts, the specialised multinucleated cells responsible for tumour osteolysis, are derived from monocyte/macrophage precursors. Breast cancer-derived tumour-associated macrophages (TAMs) are capable of osteoclast differentiation but the cellular and humoral mechanisms controlling this activity are uncertain. In this study, TAMs were isolated from primary breast cancers and cultured in the presence and absence of cytokines/growth factors influencing osteoclastogenesis. Extensive TAM-osteoclast differentiation occurred only in the presence of RANKL and M-CSF; this process was inhibited by OPG and RANK:Fc, decoy receptors for RANKL. Breast cancer-derived fibroblasts and human bone stromal cells expressed mRNA for RANKL, OPG and TRAIL, and co-culture of these fibroblasts with human monocytes stimulated osteoclast formation by a RANKL-dependent mechanism. Osteoclast formation and lacunar resorption also occurred by a RANKL-independent mechanism when the conditioned medium from breast cancer cells, MDA-MB-231 and MCF-7, was added (with M-CSF) to monocyte cultures. Our findings indicate that TAMs in breast cancer are capable of osteoclast differentiation and that breast cancer-derived fibroblasts and breast cancer cells contribute to this process by producing soluble factors that influence osteoclast formation by RANKL-dependent and RANKL-independent mechanisms respectively.     

纤维连接蛋白在不同类型肺癌细胞侵袭过程中的作用及机制

目的 观察和探讨纤维连接蛋白(FN)对不同类型肺癌细胞侵袭能力的影响及机制。方法 在FN包被的培养板和FN滤膜的Boyden浸润小室等体外侵袭模型中，比较小细胞肿癌细胞系054A和肺腺癌细胞系A549粘附及迁移能力的差别，并检测FN对肿瘤细胞增殖活性的影响。分别给予细胞抗α3整合素、抗α5整合素、抗β1整合素单抗预处理后，观察侵袭性的变化。结果 FN增强A549细胞粘附、迁移及增殖活性的作用明显大于054A细胞。这种作用在A549细胞能被抗α3、抗α5和抗β1单抗抑制，而在054A细胞能被抗α3和抗β1单抗抑制。结论 FN对不同类型肺癌细胞侵袭能力的影响不同，可能与细胞膜整合素受体的差异有关。