诱导化疗在舌鳞状细胞癌治疗中的应用及意义

目的探讨诱导化疗对舌鳞癌患者局部病灶及生存率的影响。方法 63例舌鳞癌患者随机分为2组:Ⅰ组(单纯手术)42例;Ⅱ组(诱导化疗+手术)21例,应用SPSS10.0软件统计分析2组5年生存率。结果 (1)Ⅱ组诱导化疗总有效率为71.0%(22/31),完全缓解2例。(2)Ⅰ、Ⅱ组的5年生存率分别为35.7%、61.3%,2组间有显著性差异(P<0.05);Ⅱ组诱导化疗有效者和无效者5年生存率分别为68.2%、22.2%,2组间有显著性差异(P<0.05);Ⅱ组中诱导化疗无效者和Ⅰ组5年生存率分别为22.2%、35.7%,2组间无显著性差异(P>0.05)。结论有效的诱导化疗能缩小原发灶,显著提高舌鳞癌患者的生存率。     

白藜芦醇诱导人皮肤鳞状细胞癌凋亡的蛋白质组学研究

为了探讨白藜芦醇抗皮肤鳞状细胞癌生物效应和分子机制,采用MTT法测定细胞活性,细胞流式法检测细胞凋亡率;SILAC蛋白质组学方法鉴定白藜芦醇调控的蛋白质分子。白藜芦醇呈浓度依赖性抑制皮肤鳞状细胞癌细胞的活性,细胞的存活率从100%降为56.7%;50μg/mL白藜芦醇处理导致37.3%细胞发生凋亡;鉴定了11个受白藜芦醇调控的蛋白质分子,其中BAG1,PDCD11,BCLAF1,HSPA9,YWHAZ等5个细胞凋亡相关蛋白被发现受白藜芦醇调控。     

Smac和MDM2蛋白在子宫颈鳞状细胞癌中的表达及意义

目的探讨Smac和MDM2蛋白的表达与宫颈鳞癌临床病理参数之间的关系,以了解其在宫颈癌的发生、发展、转移中的作用及意义。方法用免疫组织化学方法,检测60例宫颈鳞癌和45例宫颈上皮内瘤变(CIN)及15例正常宫颈组织中smac和MDM2蛋白的表达,并分析其表达与年龄、肿瘤大小、组织学分级及淋巴结转移间的关系。结果Smac蛋白在正常宫颈、低级别宫颈上皮内瘤变(CINⅠ)、高级别宫颈上皮内瘤变(CINⅡ-Ⅲ)及宫颈鳞癌组织中的阳性表达率分别为100%(15/15)、85%(17/20)、72%(18/25)及65%(39/60)。MDM2蛋白在正常宫颈、子宫颈低、高级别鳞状上皮内瘤变及宫颈鳞癌中的阳性表达率分别为0(0/15)、10%(2/20)、22.4%(6/25)及61.7%(37/60);Smac和MDM2蛋白的表达在正常宫颈对照组和CINII-III及与宫颈鳞癌之间均有统计学差异(P<0.05)。宫颈鳞癌中Smac和MDM2蛋白表达在淋巴结有无转移及组织分化分组中,差异有统计学意义(P<0.05);Smac蛋白和MDM2蛋白的表达之间存在负相关性(r=-0.435,P<0.001)。结论Smac和MDM2...     

PLCH1基因在食管鳞状细胞癌中的表达及其对细胞增殖、迁移的影响

目的 检测PLCH1基因在食管鳞状细胞癌（esophageal squamous cell carcinoma,ESCC）中的变异及表达情况,分析PLCH1在食管癌中的功能,并初步探讨其作用机制。方法 利用GISTIC分析PLCH1在ESCC中的拷贝数变异情况,TCGA数据库及免疫组织化学法检测PLCH1在ESCC与正常食管组织中的表达情况。通过实时荧光定量PCR(qPCR)和Western blot检测ESCC细胞系中PLCH1的表达情况。采用MTT、克隆形成试验、Transwell法等检测PLCH1沉默对ESCC细胞增殖及迁移能力的影响。结果 PLCH1在ESCC中存在显著的拷贝数扩增（G-scores> 0.1,P <0.05）,ESCC中PLCH1 mRNA及蛋白表达水平均显著高于正常组织（F=36.00～1 101.00,P均<0.000 1）。PLCH1沉默后,ESCC细胞KYESE180和TE-9的增殖能力、克隆形成能力及迁移能力均显著降低（F=35.49～634.00,P均<0.001）。结论 PLCH1在ESCC中发挥癌基因作用,对ESCC的转...     

RNA干扰整合素连接激酶基因表达对人舌鳞癌Tb细胞生长、迁移和侵袭能力的影响

目的探讨沉默整合素连接激酶(integrin-linked kinase,ILK)基因表达对人舌鳞癌Tb细胞生长、迁移和侵袭能力的影响。方法将ILK siRNA表达质粒和阴性对照质粒在脂质体介导下转染人舌鳞癌Tb细胞,稳定表达细胞株分别命名为Tb siILK和Tb vector组,并设正常Tb细胞对照组(Tb组)。Western blot法检测细胞中ILK蛋白的表达;细胞免疫荧光法检测细胞中ILK、p-Akt和p-GSK3β的表达;光镜和HE染色观察细胞的形态学变化;划痕试验检测细胞的迁移能力;Transwell法检测细胞的侵袭能力;流式细胞术检测细胞的细胞周期和凋亡情况;MTT法检测细胞的增殖活力;Western blot法检测沉默ILK基因后细胞中p-Akt、Akt、p-GSK3β、GSK3α/β、Snail的表达。结果与Tb和Tb vector组相比,Tb siILK组细胞中ILK蛋白的表达水平显著降低(P<0.01);ILK、p-Akt、p-GSK3β在胞质中的荧光信号明显减弱;大部分细胞的上皮形态特征更为明显;细胞的迁移距离和侵袭细胞数显著减少(P<0.05);G2-M期和G0-G1期细胞比例均显著增加(P<0.01);细胞的增殖活力显著减低;ILK基因沉默后,细胞中p-Akt、p-GSK3β和Snail蛋白的表达水平显著降低(P<0.05)。3组细胞的凋亡率差异无统计学意义(P>0.05)。结论抑制ILK基因的表达可通过Akt/GSK3β/Snail途径显著抑制人舌鳞癌Tb细胞的生长、迁移和侵袭能力,ILK有望作为治疗人舌鳞癌的靶基因。     

GSTθ转染对宫颈癌HeLa细胞侵袭性的影响

目的 了解增加GSTθ表达水平对人类宫颈癌HeLa细胞生长及侵袭能力的影响。方法 通过脂质体将pcDNA3-GSTθ真核质粒载体转染获得高GSTθ表达水平的HeLa稳定转染细胞;采用RT-PCR检测GSTθmRNA的表达水平;采用细胞记数和MTT法测定细胞的生长;采用划痕拍照方法观察细胞侵袭能力的改变。结果 GSTθ高水平表达显著地延缓和降低HeLa细胞的生长,并使HeLa细胞的侵袭能力下降。结论 GSTθ高水平表达可以延缓和降低细胞生长并降低HeLa细胞的侵袭能力。     

骨形态发生蛋白9对食管鳞状细胞癌细胞的抑制作用

目的探讨骨形态发生蛋白9(Bone morphogenetic proteins 9,BMP9)对食管鳞状细胞癌细胞增殖、克隆、迁移、侵袭及细胞周期的影响。方法用AdGFP和AdBMP9分别感染3株食管鳞状细胞癌细胞株ECA109、KYSE150和KYSE180,并设不感染病毒的3种细胞作为对照,采用RT-PCR法检测AdBMP9感染的3株细胞中BMP9基因mRNA的转录水平,MTT法检测病毒感染细胞0⁹6 h各组细胞的增殖活力,平板克隆法检测各组细胞的克隆形成能力,划痕试验检测各组细胞的迁移能力,Transwell小室试验检测各组细胞的侵袭能力,并对3组ECA109细胞进行细胞周期的检测。结果 AdBMP9感染的3株细胞中BMP9基因mRNA的转录水平均明显提高。与AdGFP感染和未感染病毒的细胞相比,3种细胞在过表达BMP9后,增殖活力明显降低(P<0.05);细胞克隆形成能力均下降(下降率达70%以上),且克隆细胞团明显变小;细胞迁移和侵袭能力均下降;AdBMP9感染的ECA109细胞处于G1期的细胞比例明显上升,S期比例明显下降(P<0.05)。结论 BMP9可明显抑制食管鳞癌细胞的增殖、克隆、迁移和侵袭能力,并将细胞周期阻滞于G1期。     

肿瘤侵袭抑制因子-2对基因重组肝细胞生长因子诱导的人肺癌细胞运动和侵袭的抑制作用

应用基因重组肝细胞生长因子（HGF）可诱导人肺癌细胞运动和侵袭，通过细胞离散、集落扩散和基底膜侵袭实验证明，侵袭抑制因子-2（ⅡF-2）可明显地抑制由HGF诱导的作用。该抑制作用可被抗ⅡF-2抗体所阻断，表明ⅡF-2在肿瘤转移的预防和治疗方面可能具有重要意义。     

17β-雌二醇对人甲状腺滤泡状癌WRO细胞侵袭能力的影响及其机制

目的探讨17β-雌二醇(17β-estradiol,E2)对人甲状腺滤泡状癌WRO细胞侵袭能力的影响及其机制。方法用10-8mol/L的E2处理WRO细胞不同时间(0、24、48 h),倒置显微镜下观察细胞形态的变化;RT-PCR和Western blot法检测细胞中G蛋白偶联雌激素受体(G protein-coupled estrogen receptor,GPER)、CXCR1、基质金属蛋白酶2(matrix metalloproteinase2,MMP2)和MMP9基因mRNA的转录水平和蛋白的表达水平;ELISA法检测细胞中IL-8的含量;Transwell小室法检测细胞的侵袭能力。结果随着E2处理时间的延长,WRO细胞间的间隙变大,逐渐离散成单个细胞,形态呈长梭形、纺锤形;细胞中GPER、CXCR1、MMP2和MMP9基因mRNA的转录水平和蛋白的表达水平及分泌IL-8的水平均逐渐升高(P<0.05或P<0.01);细胞的侵袭能力明显增强(P<0.05)。结论 E2可通过上调GPER、CXCR1、MMP2、MMP9和IL-8的表达,促进WRO细胞的侵袭。     

Gangliosides of cultured cells of a rat mammary carcinoma cell line

Ganglioside content of rat mammary carcinoma-derived cells grown in layers in vitro was nearly as high as that of apical plasma membrane-derived milk fat globule membrane and nearly four times higher than the content of normal, lactating mammary tissue on a protein basis. The major ganglioside of these carcinoma-derived cells was identified as GDla (sialic acid-Gal-GalNAc-(sialic acid)-Gal-Glc-Cer. Relative to carcinoma-derived cells, rat mammary tissue and milk fat globule membrane had more complex ganglioside patterns but appeared to lack substantial quantities of GDla.     

Three-dimensional modeling of squamous cell carcinoma antigen 2 and its interactions with serine protease

An attempt is made to build up a three-dimensional model of squamous cell carcinoma antigen 2 (SCCA2) by means of the homology module of Insight II, where SCCA2 contains the stressed and relaxed forms, i.e. SCCA2(S) and SCCA2(R). The docking of SCCA2(S) with two different target serine proteinases, that are the cathepsin G (Cat G) and the human mast cell chymase (HMC), are studied theoretically to give two different complexes, respectively. It is shown, from the molecular surface electricity potential, that in the formation of the two complexes SCCA2(S)-Cat G and SCCA2(S)-HMC, the electrostatic interaction may play an important role. Since HMC possesses a loop to produce spatial anti-effect, the complex SCCA2(S)-HMC is less stable than the complex SCCA2(S)-Cat G. However, the reactive site loop involved in SCCA2(S) is an important factor in restraining serine proteinases.     

Microsatellite analysis in multistage carcinogenesis of esophageal squamous cell carcinoma from chongqing in southern china

In order to characterize the molecular events in the carcinogenesis of esophageal cancer and to identify biomarkers for the early detection of the disease, matched precancerous and cancerous tissues resected from 34 esophageal cancer patients in Chongqing of southern China were compared for the extent of loss of heterozygosity (LOH). Sixteen microsatellite markers on nine chromosome regions were used for the PCR-based LOH analysis. The overall frequency of LOH at the 16 microsatellite loci was significantly increased as the pathological status of the resection specimens changed from low-grade dysplasia (LGD) to high-grade dysplasia (HGD) and squamous cell carcinoma (SCC) (P < 0.001), indicating that tumorigenesis of the esophageal squamous epithelia is a progressive process involving accumulative changes of LOH. A total of eight markers showed LOH in the LGD samples, suggesting that these loci may be involved in the early-stage tumorigenesis of esophageal squamous cell carcinoma (ESCC) and that LOH analysis at these loci may help improve the early detection of this disease. In addition, heterozygosity was regained at four loci in the SCC samples of four patients compared with the HGD samples, suggesting the possibility of genetic heterogeneity in the tumorigenesis of esophageal cancer.     

Differentiation of human promyelocytic leukemia cell line HL60 by microbial extracellular glycolipids

Microbial extracellular glycolipids, succinoyl trehalose lipid (STL), and mannosylerythritol lipid (MEL) inhibited the growth of a human promyelocytic leukemia cell line, HL60, and induced their morphological changes. The results of specific and nonspecific leukocyte esterase activities showed that STL induced monocytotic differentiation while MEL induced granulocytic differentiation. STL and MEL markedly increased common differentiation-associated characteristics in monocytes and granulocytes, such as nitroblue tetrazolium (NBT) reducing ability, expression of Fc receptors, and phagocytic activities in HL60 cells, respectively. Neither sugar moieties nor fatty acids in the free form, the individual components of STL and MEL, were effective at inducing the differentiation of HL60 cells. The induction of differentiation was not due to surface activities of STL and MEL on the basis of the complete ineffectiveness of the analogues tested. The composition of cell surface glycosphingolipids (GSL) changed such that the G_M3/LacCer ratio increased in STL-treated cells, whereas it decreased in MEL-treated cells. HL60 cells treated with STL and MEL exhibited a significant decrease in the activity of the intracellular phospholipid- and Ca²⁺-dependent protein kinase (protein kinase C). Furthermore, the serine/threonine phosphorylations in intact HL60 cells were clearly inhibited by the presence of G_M3 and MEL, but not by LacCer and STL. These results suggest that the differentiation-inducing activity of STL and MEL is not due to a simple detergent-like effect but due to a specific action on the plasma membrane. The inhibitory effect of STL on protein kinase activity was through increasing G_M3, but MEL had a direct inhibitory effect.     

Characterization of immortalized MARCO and SR-AI/II-deficient murine alveolar macrophage cell lines

Background  

Alveolar macrophages (AM) avidly bind and ingest unopsonized inhaled particles and bacteria through class A scavenger receptors (SRAs) MARCO and SR-AI/II. Studies to characterize the function of these SRAs have used AMs from MARCO or SR-AI/II null mice, but this approach is limited by the relatively low yield of AMs. Moreover, studies using both MARCO and SR-AI/II-deficient (MS^-/-) mice have not been reported yet. Hence, we sought to develop continuous cell lines from primary alveolar macrophages from MS^-/- mice.     

Effect of tocotrienols on the growth of a human breast cancer cell line in culture

The tocotrienol-rich fraction (TRF) of palm oil consists of tocotrienols and some α-tocopherol (α-T). Tocotrienols are a form of vitamin E having an unsaturated side-chain, rather than the saturated side-chain of the more common tocopherols. Because palm oil has been shown not to promote chemically-induced mammary carcinogenesis, we tested effects of TRF and α-T on the proliferation, growth, and plating efficiency (PE) of MDA-MB-435 estrogen-receptor-negative human breast cancer cells. TRF inhibited the proliferation of these cells with a concentration required to inhibit cell proliferation by 50% of 180 μg/mL, whereas α-T had no effect at concentrations up to 1000 μg/mL as measured by incorporation of [³H]thymidine. The effects of TRF and α-T also were tested in longer-term growth experiments, using concentrations of 180 and 500 μg/mL. We found that TRF inhibited the growth of these cells by 50%, whereas α-T did not. Their effect on the ability of these cells to form colonies also was studied, and it was found that TRF inhibited PE, whereas α-T had no effect. These results suggest that the inhibition is due to the presence of tocotrienols in TRF rather than α-T. Based on a paper presented at the PORIM International Palm Oil Congress (PIPOC) held in Kuala Lumpur, Malaysia, September 1993.     

Macronutrients, vitamins and minerals intake and risk of esophageal squamous cell carcinoma: a case-control study in Iran

Background

Although Iran is a high-risk region for esophageal squamous cell carcinoma (ESCC), dietary factors that may contribute to this high incidence have not been thoroughly studied. The aim of this study was to evaluate the effect of macronutrients, vitamins and minerals on the risk of ESCC.

Methods

In this hospital-based case-control study, 47 cases with incident ESCC and 96 controls were interviewed and usual dietary intakes were collected using a validated food frequency questionnaire. Data were modeled through unconditional multiple logistic regression to estimate odds ratios (OR) and 95% confidence intervals (CI), controlling for age, sex, gastrointestinal reflux, body mass index, smoking history (status, intensity and duration), physical activity, and education.

Results

ESCC cases consumed significantly more hot foods and beverages and fried and barbecued meals, compared to the controls (p < 0.05). After adjusting for potential confounders, the risk of ESCC increased significantly in the highest tertiles of saturated fat [OR:2.88,95%CI:1.15-3.08], cholesterol [OR:1.53, 95%CI: 1.41-4.13], discretionary calorie [OR:1.51, 95%CI: 1.06-3.84], sodium [OR:1.49,95%CI:1.12-2.89] and total fat intakes [OR:1.48, 95%CI:1.09-3.04]. In contrast, being in the highest tertile of carbohydrate, dietary fiber and (n-3) fatty acid intake reduced the ESCC risk by 78%, 71% and 68%, respectively. The most cancer-protective effect was observed for the combination of high folate and vitamin E intakes (OR: 0.02, 95%CI: 0.00-0.87; p < 0.001). Controls consumed 623.5 times higher selenium, 5.48 times as much β-carotene and 1.98 times as much α-tocopherol as the amount ESCC cases consumed.

Conclusion

This study suggests that high intake of nutrients primarily found in plant-based foods is associated with a reduced esophageal cancer risk. Some nutrients such as folate, vitamin E and selenium might play major roles in the etiology of ESCC and their status may eventually be used as an epidemiological marker for esophageal cancer in Iran, and perhaps other high-risk regions.     

Aurora-A高表达对食管鳞癌血管生成及血管内皮生长因子受体-2表达的影响

目的探讨Aurora-A高表达对食管鳞癌血管生成及血管内皮生长因子受体-2(vascular endothelial growthfactor receptor 2,VEGFR-2)表达的影响。方法将绿色荧光蛋白(green fluorescent protein,GFP)标记的Aurora-A全长表达质粒pEGFP-C1-Aurora-A转染至食管鳞癌细胞KYSE150,经G418筛选获得Aurora-A高表达的细胞株(Aurora-A高表达组),同时设空质粒pEGFP-C1转染组(阴性对照组)和未转染组作为对照。Western blot检测各组细胞中Aurora-A蛋白的表达;采用小管形成及鸡胚尿囊膜试验检测Aurora-A高表达对肿瘤血管生成的影响;免疫组化法检测Aurora-A高表达对裸鼠移植瘤中微血管密度(microvessel density,MVD)的影响;Western blot检测Aurora-A高表达对KYSE150细胞中VEGFR-2及p-VEGFR-2(Tyr1059)表达的影响。结果 Aurora-A高表达组中总Aurora-A蛋白的表达量约为阴性对照组和未转染组的2.5倍,提示Aurora-A高表达细胞系构建成功;Aurora-A高表达组生成的血管数量、MVD值和p-VEGFR-2的表达水平均显著高于阴性对照组(P<0.01)。结论 Aurora-A高表达可促进食管鳞癌中的血管生成,其机制可能与活化VEGFR-2有关。     

Time course of incorporation of 20-carbon polyunsaturated fatty acids in a human keratinocyte cell line

Human keratinocytes (NCTC 2544) in culture were labeled with equal amounts of either¹⁴C-arachidonic acid,¹⁴C-dihomo-γ-linolenic acid or¹⁴C-eicosapentaenoic acid. At various time points, the incubations were stopped and the distribution of the¹⁴C-fatty acids was analyzed. All these eicosanoid precursor fatty acids were effectively incorporated into the cellular lipids of the keratinocytes, and the major radiolabeled individual lipid fraction was phosphatidylethanolamine. The distributions of arachidonic acid and dihomo-γ-linolenic acid within cellular lipids were rather the same. However, less eicosapentaenoic acid than either arachidonic acid or dihomo-γ-linolenic acid was incorporated into the phospholipids and, correspondingly, more eicosapentaenoic acid was incorporated into the nonphosphorus lipids. In the phosphatidylinositol + phosphatidylserine fraction, there was significantly less eicosapentaenoic acid than either arachidonic acid or dihomo-γ-linolenic acid. The present study suggests that these eicosanoid precursor fatty acids are effectively incorporated into the human keratinocytes and that the pattern of incorporation and distribution of eicosapentaenoic acid appears to differ slightly from that of either arachidonic acid or dihomo-γ-linolenic acid.