树突状细胞在肺腺癌组织中的浸润及其影响

目的：研究肺腺癌组织中树突状细胞（ＤＣ）的浸润程度及对预后的影响。方法将Ｓ－１００蛋白作为ＤＣ特异性标记物,应用免疫组织化学方法检测肺腺癌组织中ＤＣ分布。结果４６例肺腺癌中,ＤＣ显著浸润者１９例,５年生存率６１％,无或轻度浸润者２７例,５年生存率２１％,低分化腺癌２６例,显著浸润者１０例,５年生存率５７％,轻度浸润者１６例,５年生存率１６％。经Ｌｏｇ－ｒａｎｋ检验,在显著浸润和轻度浸润组二者之间差     

塞来昔布对胃癌细胞株LRP表达的影响

目的探讨塞来昔布对胃癌细胞株LRP表达的影响。方法采用MTT法检测塞来昔布不同剂量对胃腺癌细胞株SGC-7901生长的影响,在此基础上采用免疫组化及ELISA方法检测塞来昔布在一定剂量范围内某一时间点对SGC-7901细胞COX-2、LRP表达的影响。结果不同浓度的塞来昔布均对胃腺癌细胞株SGC-7901有抑制作用,并且这种影响呈时间和剂量依赖性;胃癌细胞株LRP的表达也随着塞来昔布浓度的增加而减少。结论选择性COX-2抑制剂塞来昔布可以下调胃癌细胞株中耐药基因LRP的表达,逆转多药耐药作用。     

STAT3在人肺腺癌细胞A549中的表达及意义

目的 探讨信号转导和转录激活因子3( STAT3)的临床应用价值.方法 利用电化学发光免疫法检测人肺腺癌细胞A549和人胚胎成纤维细胞MRC-5培养3d的培养液中癌胚抗原(CEA)的表达水平.采用荧光定量PCR法检测人肺腺癌细胞A549和人胚胎成纤维细胞MRC-5中STAT3mRNA、CEA mRNA的表达.结果 ...     

水通道蛋白与肺腺癌

肺癌是人类高发的恶性肿瘤,病死率在恶性肿瘤中居首位.其中,肺腺癌易发生远处转移,而目前肿瘤转移的确切机制不十分明确.最近发现水通道蛋白(aquaporin,AQP)参与细胞迁移和增殖,提示其在肿瘤生物学中的重要作用.已有研究发现,多种AQP在不同来源的恶性肿瘤中高表达,并通过促进血管新生及快速转运水分,在肿瘤细胞的迁移侵袭中起重要作用.目前国内外已有研究发现AQP1、3、5在肺腺癌细胞中高表达.深入研究肺腺癌及肺腺癌转移灶中AQP的表达可能给肿瘤细胞转移机制带来新的视角,并为将来用AQP抑制剂治疗肺腺癌、降低它的侵袭能力和远处转移提供理论依据.     

水通道蛋白1在肺腺癌细胞株A549中的表达及意义

目的 探明水通道蛋白1在人肺腺癌细胞株A549中的表达情况,以探讨水通道蛋白1作为肺腺癌治疗靶位的可能性.方法 培养肺腺癌细胞株A549,应用免疫细胞化学法、逆转录聚合酶链反应及Western blot等方法检测水通道蛋白1表达.结果 三种方法均证实肺腺癌细胞株A549存在水通道蛋白1的表达.结论 肺腺癌细胞株A549...     

二甲双胍对肺腺癌耐厄洛替尼细胞株A549ER的耐药逆转作用

目的 探讨二甲双胍对人肺腺癌耐厄洛替尼细胞株A549ER的耐药逆转作用.方法 将人肺腺癌细胞株A549细胞设为亲本组;将耐厄洛替尼细胞株A549ER细胞分为空白对照组、厄洛替尼组、二甲双胍组和联合用药组（厄洛替尼±二甲双胍组）,采用CCK8法检测不同浓度药物作用下各组细胞的50％抑制浓度（IC50）,计算耐药倍数和逆转倍数.采用流式细胞术检测各组A549ER细胞的凋亡率和细胞周期,计算增殖指数.结果 在0 ～ 20 mmol/L浓度范围内,二甲双胍对A549细胞及A549ER细胞均有生长抑制作用,抑制率随二甲双胍浓度升高而增加.厄洛替尼对A549细胞和A549ER的IC50分别为15.15 μmol/L和118.8 μmol/L,A549ER的耐药倍数为7.84.联合用药组A549ER细胞IC50为73.55 umol/L,耐药倍数为4.85.二甲双胍对A549ER厄洛替尼耐药性的逆转倍数为1.62.空白对照组、厄洛替尼组、二甲双胍组和联合用药组的凋亡率分别为（5.53±3.00）％、（7.51±3.73）％、（10.25 ±4.23）％和（16.92±1.20）％.根据细胞周期结果计算空白对照组、厄洛替尼组、二甲双胍组和联合用药组的增殖指数分别为0.84±0.15、0.78 ±0.10、0.73±0.08和0.60 ±0.09.结论 A549ER细胞较A549细胞对厄洛替尼有明显的耐药性;二甲双胍对A549ER细胞厄洛替尼的耐药性具有逆转作用;二甲双胍通过抑制细胞生长、促进细胞凋亡、减缓细胞周期进程等途径逆转A549ER细胞耐药.     

人肺腺癌细胞系A549线粒体差异蛋白质组学研究

目的 研究人肺腺癌细胞系A549与人正常支气管上皮细胞系16HBE线粒体蛋白质组的差异表达.方法 传代培养细胞系A549及16HBE,用线粒体提取试剂盒获取细胞线粒体蛋白质,进行双向凝胶电泳,运用液相色谱串联质谱分析技术筛选出A549和16HBE细胞系线粒体间表达水平显著差异的蛋白,所得结果通过Data Analysis软件标峰,用MASCOT进行结果搜索和数据分析.结果 双向电泳结果显示A549、16HBE细胞系线粒体存在差异的蛋白质点共41个,3倍以上差异的16个,A549细胞系中表达上调的15个,表达下调的26个,其中3倍以上差异表达上调的7个,表达下调的9个.运用液相色谱串联质谱技术鉴定出A549细胞系线粒体表达上调的蛋白质2个:AAA+ ATP酶家族结构域蛋白3B、tRNA鸟嘌呤糖基转移酶,表达下调的蛋白质7个:热休克蛋白75、复合物Ⅲ亚基1、复合物Ⅲ亚基2、鸟氨酸氨基转移酶、异柠檬酸脱氢酶亚基α、SLP-2、抗增殖蛋白.结论 应用亚细胞蛋白质组学方法,鉴定出肺腺癌细胞系线粒体差异表达蛋白,为阐明肺腺癌发生的分子机制、筛选早期诊断标志物提供了有益的线索.     

EGLN3在肺腺癌中表达的Meta分析

目的 分析EGLN3在肺腺癌(lung adenocarcinoma, LUAD)中的作用。方法 (1)通过癌症基因组图谱(the cancer genome atlas, TCGA)数据库下载535例肺腺癌样本和59例正常样本,分析EGLN3在肺腺癌和正常肺组织中的表达差异;(2)使用R语言分析EGLN3的表达水平及其在肺腺癌中的临床意义;(3)通过Kaplan-Meier方法、单因素和多因素COX回归分析和生存预测列线图确定EGLN3的表达水平与肺腺癌预后的关系;(4)通过基因集富集分析(GSEA)筛选EGLN3相关的生物学途径;(5)细胞实验：通过蛋白质印迹法(WB)验证EGLN3在肺腺癌细胞中是否存在差异表达;对EGLN3基因进行过表达和干扰表达,采用CCK-8增殖实验、划痕实验和Transwell侵袭实验,明确表达水平对肺腺癌细胞增殖、迁移和侵袭的影响。结果 (1)EGLN3在肺腺癌组织中表达水平高于正常肺组织细胞;(2)EGLN3表达水平与肺腺癌病理分期相关,病理分期越高,对应EGLN3表达水平越高;(3)EGLN3表达水平与肺腺癌总生存期(OS)独立预后相关,EGLN3高...     

人子宫内膜腺癌细胞系的建立

目的构建人子宫内膜腺癌细胞系。方法将1例高分化子宫内膜腺癌手术切除的肿瘤组织制成单细胞悬液,用含10%胎牛血清的DMEM/F-12培养基进行原代培养,倒置显微镜下观察细胞形态,并用免疫荧光染色法检测其角蛋白。结果体外培养细胞传至50代以上,传代后细胞单层贴壁,散在生长,密度增加时见堆积现象,失去接触抑制,细胞质中角蛋白表达阳性。结论成功建立了人子宫内膜腺癌细胞系。     

人呼吸道合胞病毒转录调节基因转染肺腺癌细胞系的研究

目的获得稳定表达人呼吸道合胞病毒(hRSV)M2-1基因的人肺腺癌细胞系.方法通过基因重组法构建hRSV M2-1基因真核表达载体,用脂质体将其转染人肺腺癌PAa和A549细胞,经G418筛选获得阳性表达细胞株,并用逆转录-聚合酶链反应(RT-PCR)和Western Blot进行验证.结果得到了约650bp的基因插入片段,DNA测序表明该基因高度保守.筛选出了稳定高量表达M2-1基因的PAa和A549细胞,并被证实有M2-1蛋白表达.结论获得了稳定表达hRSV M2-1蛋白的PAa和A549细胞株.     

Inhibitor of growth 4 induces apoptosis in human lung adenocarcinoma cell line A549 via Bcl-2 family proteins and mitochondria apoptosis pathway

Objective  Inhibitor of growth 4 (ING4) is considered to be a tumor suppressor implicated in several human malignancies by tumor growth inhibition and apoptosis enhancement. In present study, the effects of ING4 on apoptosis and its mechanisms were investigated through the transduction of ING4 cDNA into lung adenocarcinoma cell line A549. Methods  The effects of ING4 on A549 apoptosis were observed by FCM analysis, TUNEL assay, and electron microscopy. Simultaneously, the effects of ING4 on the expression of several apoptosis-related proteins in cell line A549 were evaluated by Western blot analysis. Results  Both Annexin-V FITC analysis by FCM and TUNEL assay revealed more apoptotic cells in A549 cells with exogenous ING4 gene. For electron microscopy, A549 cells with exogenous ING4 gene showed typical morphological changes of apoptosis. The deregulation of Bcl-2 family proteins (Bcl-2, Bcl-xl, Bax, Bak, Bid) and the major apoptotic executioners of mitochondria pathway (Cyt-c, caspase3, PARP) were also observed. Conclusion  Our findings suggest that exogenous ING4 can enhance A549 apoptosis via regulating the expression of Bcl-2 family proteins and the activation of mitochondrial apoptotic pathway. Xiaomei Li, Qingyuan Zhang, and Limin Cai contributed equally to this work.     

Coincidence between malignant perivascular epithelioid cell tumor arising in the gastric serosa and lung adenocarcinoma

A 4-mo history of both epigastralgia and back pain was presented in a 39-year-old male. Computed tomography showed right lung nodule and abdominal mass attached to the gastric wall, measuring approximately 30 mm and 70 mm in diameter. Since biopsy samples from the lung and abdomen revealed poorly differentiated adenocarcinoma and malignant tumor, clinicians first interpreted the abdominal mass as metastatic carcinoma, and a right lower lobectomy with following resection of the mass was performed. Gross examination of both lesions displayed gray-whitish to yellow-whitish cut surfaces with hemorrhagic and necrotic foci, and the mass attached to the serosa of the lesser curvature on the gastric body. On microscopic examination, the lung tumor was composed of a proliferation of highly atypical epithelial cells having abundant eosinophilic cytoplasm, predominantly arranged in an acinar or solid growth pattern with vessel permeation, while the abdominal tumor consisted of sheets or nests with markedly atypical epithelioid cells having pleomorphic nuclei and abundant eosinophilic to clear cytoplasm focally in a radial perivascular or infiltrative growth pattern. Immunohistochemically, the latter cells were positive for HMB45 or α-smooth muscle actin, but the former ones not. Therefore, we finally made a diagnosis of malignant perivascular epithelioid cell tumor(PEComa) arising in the gastric serosa, combined with primary lung adenocarcinoma. Furthermore, small papillary car-cinoma of the thyroid gland was identified. The current case describes the coincidence of malignant PEComa with other carcinomas, posing a challenge in distinction from metastatic tumor disease.     

Translational research on esophageal adenocarcinoma: from cell line to clinic

Human esophageal adenocarcinoma (EAC) cell lines have made a substantial contribution to elucidating mechanisms of carcinogenesis and drug discovery. Model research on EAC relies almost entirely on a relatively small set of established tumor cell lines because appropriate animal models are lacking. Nowadays, more than 20% of all fundamental translational research studies regarding EAC are partially or entirely based on these cell lines. The ready availability of these cell lines to investigators worldwide have resulted in more than 250 publications, including many examples of important biomedical discoveries. The high genomic similarities (but certainly not completely identical) between the EAC cell lines and their original tumors provide rational for their use. Recently, in a collaborative effort all available EAC cell lines have been verified resulting in the establishment of a reliable panel of 10 EAC cell lines. It could be expected that the value of these cell lines increases as unlimited source of tumor material because new biomedical techniques require more tumor cells and the supply of viable tumor cells is diminishing because of neoadjuvant chemo(radio)therapy of patients with EAC. Here, we review the history of the EAC cell lines and their utility in translational research and biomedical discovery.     

肺腺癌经气道播散肺内转移

肺腺癌经气道播散肺内转移是指肺腺癌细胞通过气道扩散植入同侧或对侧其他肺段或肺叶,形成新的肺内转移癌灶,是肺特有的一种恶性肿瘤转移机制.了解肺腺癌经气道播散肺内转移可能的发病机制、病理学和影像特征,有助于提高对其认识,加强早期干预.     

TNP-470对肺腺癌细胞增殖和凋亡的影响

目的 探讨TNP－470对肺腺癌细胞增殖和凋亡的影响及其作用机制。方法 以不同浓度的TNP－470信息处理地培养的肺腺癌AGZY－82A细胞株,以免疫组化S－P法检测肺腺癌细胞增殖核抗原（PCNA）、p53、bcl-2、血管内皮生长因子（VEGF）和VEGF受体Flk-1的表达。结果 随着TNP－470浓度的增加,肺腺癌细胞VEGF、Flk-1、PCNA的表达减少,p53、bcl-2的表达逐渐增加;当TNP－470为10^7μg/L时,细胞增殖几乎停止,随着TNP－470（10^4μg/L）作用时间的延长,肺腺癌细胞VEGF、Flk-1、PCNA的表达逐渐减少,而p53、bcl-2的表达逐渐增加。结论 TNP－470通过使肺腺癌细胞自分泌VEGF减少,Flk表达降低,从而抑制肺腺癌细胞增殖,促进其凋亡。     

CEACAM5在肺腺癌中的表达及临床意义

目的 通过检测癌胚抗原相关细胞黏附分子5(CEACAM5)在肺腺癌中的表达,探究其临床意义及预后.方法 回顾性分析2016年8月-2017年8月在本院确诊为肺腺癌的患者51例,取肺腺癌患者病变肺部组织作为实验组;从中选取42例取癌旁组织作为对照组.采用免疫组化SP法检测肺部肺腺癌组织和癌旁组织CEACAM5的表达,并利...     

小剂量顺铂和5-氟尿嘧啶联合化疗治疗晚期肺腺癌

目的　观察小剂量顺铂和 5 -氟尿嘧啶联合方案 (FP方案 )治疗晚期肺腺癌的疗效及毒副作用。方法　 FP方案 5 -氟尿嘧啶 32 0 mg·m2 - 1 · d- 1 ,持续 8h,连用 2周 ,DDP5 mg·m2 - 1 · d- 1每周 5日连用 2周 ,其后休息 2周 ,连用两个周期后进性疗效评价。结果　小剂量 FP方案治疗晚期肺腺癌 2 0例 ,总有效率 35 %。主要毒副作用为轻度的血液和消化道副反应。结论　小剂量 FP化疗方案是治疗晚期肺腺癌有效且毒性较小的联合化疗方案。     

XXYLT1 methylation contributes to the occurrence of lung adenocarcinoma: Methylation and lung adenocarcinoma

Background:There is evidence that DNA methylation play major roles in lung cancer. In our previously study, C3 or f21 , also referred to as XXYLT1, rs2131877 polymorphism is associated with a reduced risk of lung adenocarcinoma. So, we explored the role of XXYLT1 methylation in lung adenocarcinoma.Methods:This study was conducted in 2 steps. In the first step, we recruited 15 patients with lung adenocarcinoma. Cancer tissues and para-carcinoma tissues were obtained from each of the patients. In the second step, 150 patients with lung adenocarcinom were enrolled, and cancer and normal lung tissue were obtained from each patients, respectively. The expression levels of XXYLT1 mRNA were determined, the deoxyribonucleic acid methylation status was analyzed by MassARRAY Spectrometry. The methylation data of individual units were generated by EpiTyper v1.0.5 software.Results:The XXYLT1 mRNA expression was significantly lower in cancer tissues than in para-carcinoma and normal lung tissues. Meanwhile, the methylation rates of three CpG units (CpG_23, CpG_25, and CpG_60.61.62.63.64.65) within the XXYLT1 gene were higher in cancer tissues compared to the para-carcinoma and the normal lung tissues. This difference was particularly significant in male patients.Conclusions:Our results suggested that methylation of XXYLT1 may have significance in the pathogenesis of lung adenocarcinoma.     

NCAPG stimulates lung adenocarcinoma cell stemness through aerobic glycolysis

Background

Cancer stem cells are pivotal in cancer progression and therapy, including lung adenocarcinoma (LUAD). High NCAPG level is implicated in malignant tumorigenesis, but investigations on NCAPG and LUAD stem cells are warranted. Hence, projecting the impact of NCAPG on cell stemness and the targeted therapy for LUAD is of the essence.

Methods

Bioinformatics analyzed NCAPG expression in LUAD tissues. qRT-PCR assayed NCAPG expression in LUAD cells. CCK-8 assessed cell viability and cell sphere-forming assay measured sphere-forming ability. Western blot assessed expression of stem cell-related markers (CD133, CD44, Oct-4) and specific genes (HK2, PKM2, LDHA) related to glycolysis metabolism pathway. Cellular glycolytic capacity was assayed by glycolytic metabolites pyruvic acid, lactate, citrate, and malate assay kits, and extracellular acidification rate and oxygen consumption rate analyzers.

Results

NCAPG was upregulated in LUAD and enriched in the aerobic glycolysis pathway, and its expression was positively correlated with that of glycolytic marker genes. Cell function assays revealed that NCAPG stimulated proliferation, stemness, and glycolytic activity of LUAD cells. Rescue experiments unveiled that 2-DG (glycolysis inhibitor) was able to reverse the stimulative impact of NCAPG overexpression on proliferation, stemness, and glycolytic activity of LUAD cells.

Conclusion

NCAPG stimulated LUAD cell stemness through activation of glycolysis pathway. NCAPG may be possible biomarker for diagnosis and target for treatment of LUAD.