CD44v3短发夹RNA对透明质酸诱导人结肠癌SW480细胞体外粘附和侵袭的抑制作用

目的:研究CD44v3的短发夹RNA(short hairpin RNA,shRNA)对体外培养的人结肠癌SW480细胞CD44v3表达及透明质酸诱导的粘附侵袭行为的抑制作用。方法:设计和构建带有U6启动子的CD44v3基因特异性的shRNA干扰表达质粒,转染至SW480细胞,以RT-PCR和Western blot检测SW480细胞转染前后CD44v3表达,以平板粘附模型和Boyden小室模型检测SW480细胞转染前后粘附和侵袭能力。结果:和转染前相比,转染了RNA干扰表达质粒的SW480细胞CD44v3mRNA和蛋白表达、以及受透明质酸诱导而粘附于平板和穿过Boyden小室隔膜的细胞数皆显著减少(P<0.01)。结论:针对CD44v3的短发夹RNA能显著抑制透明质酸诱导人结肠癌SW480细胞体外粘附和侵袭行为。     

CCR7活化与人结肠癌SW620细胞的体外增殖、趋化与侵袭的关系

目的研究CCR7活化对结肠癌SW620细胞体外增殖、趋化与侵袭活性的影响。方法MTT法和软琼脂细胞集落培养观察CCR7活化对细胞增殖的影响,Boyden小室法检测CCR7活化对SW620细胞趋化和侵袭活性的影响。结果和对照组相比CCL21组细胞增殖数量、软琼脂细胞集落数和穿过Boyden小室膜的细胞数均显著增加（P〈0．01）。结论CCR7活化能够促进结肠癌SW620细胞的体外增殖、趋化与侵袭,其可能参与了结肠癌淋巴结转移的过程。进一步研究CCR7在结肠癌中的作用将有助于阐明结肠癌淋巴结转移的机制。     

AP-2α基因转染对人类结肠癌SW480细胞体外增生及侵袭能力的影响

目的 探讨AP-2α基因对人类结肠癌SW480细胞体外增生及侵袭能力的影响.方法 构建PODNA3.1(+)-AP-2α真核表达载体,利用脂质体介导pcDNA3.1(+)-AP-2α和pcDNA3.1(+)转染5W480细胞,并以正常SW480细胞作为空白对照;采用RT-PCR与Western blotting分别检测转染48 h后各组细胞中AP-2αmRNA与蛋白的表达情况;采用平板克隆、软琼脂克隆形成试验以及Transweil侵袭试验,分别检测各组细胞体外增生及侵袭能力.结果 SW480细胞内源性AP-2α蛋白表达缺失;转染AP-2α基因后,在SW480细胞中可检测到高水平的AP-2αmRNA及蛋白,细胞克隆形成率降低(P<0.05),软琼脂克隆体积小且数量少,细胞侵袭能力下降(P<0.05).结论 转染AP-2α基因可以抑制人类结肠癌SW480细胞的体外恶性增生以及侵袭能力.     

阿苯达唑抑制结肠癌SW480细胞的侵袭和迁移能力及其可能的机制

目的：探索阿苯达唑（albendazole）抑制结肠癌SW480细胞侵袭和迁移能力及其可能的机制。方法：体外培养人结肠癌SW480细胞,用不同质量浓度(0、1.0和2.0 mg/ml)的albendazole处理人结肠癌SW480细胞,CCK-8法检测albendazole对结肠癌SW480细胞增殖能力的影响,采用细胞划痕实验、Transwell小室实验检测细胞的迁移侵袭能力,免疫细胞化学及Western blotting检测SW480细胞中E-cadherin、MMP-2和MMP-9蛋白的表达水平。结果：与空白对照组比较,albendazole各质量浓度（1.0和2.0 mg/ml）组细胞的增殖能力明显降低,SW480细胞侵袭细胞数显著下降\[ （51.33±3.96）、（23.42±403）vs （80.76±7.18）个/视野,F=3.975, P=0.026\];而且细胞迁移能力显著下降\[（9.6±1.13）、（6.4±0.81）vs （19.6±1.41） mm;F=5.012, P=0.023\];E-cadherin蛋白表达水平上调,MMP-2、MMP-9蛋白表达水平明显下调。结论：Albendazole能显著抑制SW480细胞增殖,并通过上调E-cadherin的表达水平和下调MMP-2和MMP-9的分泌水平抑制细胞的侵袭和迁移能力。     

前列腺素脱氢酶对大肠癌SW480细胞恶性增殖的抑制作用

目的研究15-羟基前列腺素脱氢酶(15-hydroxyprostaglandin dehydrogenase,PGDH)对SW480大肠癌细胞恶性增殖的抑制作用及机制。方法构建PGDH真核表达质粒(pcDNA3.1-PG-DH),在大肠癌SW480细胞中转染并表达PGDH蛋白,采用MTT法、平板集落和软琼脂克隆形成实验,分析PGDH对SW480恶性增殖的影响。Westernblot检测p53、p21蛋白表达改变情况。结果转染表达PGDH后,细胞增殖受到抑制,实验组平板集落形成率为16%,而对照组为58%,两者间差异显著(P<0.05);肿瘤细胞的停泊-非依赖能力降低,实验组软琼脂克隆体积小且形成个数(7±1.68)明显少于对照组(16.3±3.63),差异显著(P<0.05)。Westernblot分析表明,转染表达PGDH后伴随着p53、p21蛋白表达升高。结论转染表达PGDH蛋白能够部分逆转大肠癌SW480细胞的恶性表型,其过程可能与p53、p21通路密切相关。     

芹菜素对人结肠癌细胞SW480增殖抑制作用的实验研究

目的:观察芹菜素对体外培养的人结肠癌SW480细胞的增殖抑制作用.方法:观察不同浓度[(10、30、60、90)μmol/L]的芹菜素在(24、48、72)h对人结肠癌SW480细胞增殖的影响:光学显微镜和电子显微镜观察芹菜素处理后人结肠癌SW480细胞形态结构的变化;运用四唑盐比色法(MTY)检测芹菜素对人结肠癌SW480细胞生长的影响.结果:经芹菜素处理的人结肠癌SW480细胞数量减少,部分细胞体积缩小,细胞膜完整,胞浆浓缩,核染色质固缩,细胞核碎裂,形成凋亡小体;MTT法检测显示芹菜素对人结肠癌SW480细胞的增殖有抑制作用,其抑制作用随着作用浓度的增加和作用时间的延长而增强.结论:芹菜素对人结肠癌SW480细胞的增殖有抑制作用,是一种高效低毒的药物.     

芬太尼对结肠癌细胞株SW480增殖及侵袭的影响

目的：探讨不同浓度的芬太尼对结肠癌细胞株 SW480增殖和侵袭的影响。方法：选用不同浓度的芬太尼（0．5、5、50ng／ml）干预结肠癌细胞株 SW480,采用四甲基偶氮唑蓝（MMT）法检测细胞增殖,Transwell法测定细胞侵袭。结果：各组芬太尼孵育 SW480细胞48h 及72h 后,均能显著抑制细胞的增殖（P ＜0．05）,且呈浓度和时间依赖性。芬太尼孵育 SW480细胞48h,与对照组相比,各浓度组均浓度依赖性抑制凋亡（P＜0．05）。结论：芬太尼可抑制结肠癌细胞株 SW480的增殖及侵袭。     

芹菜素对人结肠癌细胞株SW480体外侵袭能力的抑制作用

目的:观测芹菜素对大肠癌细胞株SW480体外侵袭、粘附和运动能力的影响。方法:人工重组基底膜技术观察药物对细胞侵袭、运动和粘附的影响。结果:芹菜素处理后SW480细胞的侵袭、运动及粘附能力下降,并呈时间和剂量依赖性。结论:芹菜素多个层面抑制SW480细胞的侵袭、运动和粘附。     

Akt抑制剂MK-2206在缺氧环境下对结肠癌SW480细胞增殖、侵袭的影响

目的:探讨Akt抑制剂MK-2206在缺氧环境下对人结肠癌SW480细胞增殖、侵袭的影响。方法:根据CCK-8实验结果选择CoCl2和MK-2206的浓度,最后分成空白组、MK-2206组、CoCl2组、MK-2206+CoCl2组;Transwell小室实验检测四组细胞的迁移和侵袭能力;RT-PCR法检测各细胞组中Akt、mTOR、HIF-1α的mRNA表达水平;Western blot技术检测各细胞组中Akt、p-Akt、mTOR、p-mTOR、HIF-1α蛋白表达的差异性情况。结果:CoCl2诱导的缺氧环境可以促进SW480细胞的侵袭、迁移(P＜0.05),同时能够促进HIF-1α的mRNA和蛋白表达(P＜0.05),但是低浓度范围内的CoCl2对SW480细胞增殖活性无明显影响(P＞0.05);MK-2206可以在体外的常氧和缺氧环境中抑制SW480细胞的增殖活性和侵袭、迁移能力(P＜0.05);MK-2206能在体外的常氧环境下抑制SW480细胞的Akt、mTOR、HIF-1α的mRNA表达(P＜0.05),缺氧环境无明显作用;同时MK-2206能够在常氧和缺氧环境中显著降低p-Akt、p-mTOR、HIF-1α的蛋白表达水平(P＜0.05)。 结论:Akt抑制剂MK-2206可以在体外的常氧和缺氧环境下抑制SW480细胞的增殖活性和侵袭能力,但缺氧环境可能会弱化MK-2206对SW480细胞迁移的抑制作用。     

shRNA干扰BAMBI 基因对人结肠癌SW480 细胞恶性生物学行为的影响

[摘要] 目的: 探讨shRNA 干扰骨形成蛋白和激活素的穿膜抑制剂(bone orphogenetic protein and activin membrane bound inhibitor, BAMBI)基因对人结肠癌SW480 细胞增殖、凋亡、侵袭和迁移的影响及其作用机制。方法: 转染SW480 细胞sh-BAMBI成功后,实时定量PCR（qRT-PCR)和Western blotting 检测BAMBI mRNA和蛋白表达水平,MTT法检测SW480 细胞增殖能力,Hoechst33258 染色检测细胞凋亡情况,Transwell 实验检测细胞侵袭能力,划痕实验检测SW480 细胞迁移能力,Western blotting检测TGF-β/Smad2 通路相关蛋白的表达水平。结果: 转染成功后sh-BAMBI组中BAMBI的mRNA和蛋白水平均低于对照组(P<0.05);与对照组相比,sh-BAMBI 组细胞增殖率明显降低(P<0.05)、细胞凋亡率显著升高(P<0.01),同时其侵袭和迁移能力明显减弱(均P<0.05)。sh-BAMBI组TGF-β 蛋白水平和p-Smad2/Smad2 比值明显高于对照组(P<0.05)。结论: shRNA干扰BAMBI可诱导人结肠癌SW480 细胞凋亡并抑制细胞增殖、侵袭和迁移,其机制可能与激活TGFβ/Smad2 通路有关。     

外泌体在肿瘤发展中的研究进展

外泌体是细胞经过"内吞-融合-外排"等一系列调控过程而形成的细胞外纳米级小囊泡。外泌体可以携带蛋白,运送RNA,在细胞间物质和信息转导中起重要作用。外泌体可能通过调控免疫功能,促进肿瘤血管新生和肿瘤转移,以及直接作用于肿瘤细胞等途径,影响肿瘤的进展。外泌体可应用于肿瘤的诊断。本文总结了近年来有关外泌体在肿瘤发展中作用的研究进展。     

Inhibitory effect of suramin in rat models of angiogenesis in vitro and in vivo

The aim of the present study was to test the ability of the chemotherapeutic agent suramin to inhibit angiogenesis in experimental models in vitro and in vivo. In the culture of rat aortic rings on fibronectin, suramin dose-dependently inhibited vascular cell growth, achieving the maximal effect (mean − 88% versus controls, P < 0.05) at 400 μg/ml. Image analysis showed that suramin could inhibit microvessel sprouting in fibrin from rat aortic rings as evaluated by the ratio between the cellular area and the mean gray value of the sample (sprouting index); suramin at 50 μg/ml significantly reduced the sprouting index from the control value of 0.35 ± 0.04 to 0.14 ± 0.02 mm²/gray level (P < 0.05). Likewise, the area occupied by cells was 19.2 ± 1.8 mm² as compared with 41.8 ± 4.2 mm² in controls (P < 0.05). In the rat model of neovascularization induced in the cornea by chemical injury, suramin at 1.6 mg/eye per day reduced the length of blood vessels (0.7 ± 0.1 mm as compared with 1.5 ± 0.1 mm in controls, P < 0.05). In the same model the ratio between the area of blood vessels and the total area of the cornea (area fraction score) was decreased by suramin from 0.19 ± 0.02 in controls to 0.03 ± 0.003 (P < 0.05). Suramin given i.p. at 30 mg/kg per day markedly inhibited the neovascularization induced in the rat mesentery by compound 48/80 or conditioned medium from cells secreting the angiogenic protein fibroblast growth factor-3 (FGF-3). The area fraction score in control rats treated with compound 48/80 was 0.31 ± 0.03, and this was reduced to 0.07 ± 0.01 by suramin (P < 0.05). After i.p. administration of FGF-3 the area fraction score was reduced by suramin from 0.29 ± 0.03 to 0.05 ± 0.01 (P < 0.05). These results provide evidence that suramin exerts inhibitory effects on angiogenesis in both in vitro and in vivo models. Received: 9 January 1998 / Accepted: 29 June 1998     

重组人血管内皮抑制素治疗恶性肿瘤的研究进展

重组人血管内皮抑制素(恩度)是一种广谱的抗血管生成分子靶向药物,主要循证证据为联合化疗治疗晚期非小细胞肺癌(NSCLC).近年来,重组人血管内皮抑制素用于治疗多种恶性肿瘤的研究逐渐增多,并取得了较好的疗效.此外,有关重组人血管内皮抑制素联合治疗手段、给药途径、给药方法的研究逐渐开展,有利于其合理应用.     

Effect of trauma of implantation of metastatic tumor in bone in mice

We studied the influence of surgical trauma to the iliac bone on the implantation of I. V. injected tumor cells, which formed tumor in the surgical wounds of 27/84 mice (32%). None of these mice or nonsurgical mice developed tumor in the opposite or uninjured pelvic bone (P < 0.0001). When different numbers (10⁵, 5 × 10⁵, and 10 × 10⁵) of TA3Ha cells were injected I. V. immediately after surgery, the frequency of tumor formation showed an increase (respectively, 32%, 63%, 71%). As the interval between induction of trauma and tumor cell injection was increased from 0 to 15 days, the frequency of tumor formation declined from 32% to 0%. These results suggest that the healing wound is a privileged site for experimental metastasis, particularly in the early stages. It is likely that the proteins in the blood clotting cascade are involved in local tumor implantation. © 1994 Wiley-Liss, Inc.     

胶质瘤中微RNA研究进展

微RNA(microRNA,miR)可在转录后水平负调控靶基因表达,miR异常表达与肿瘤生成密切相关.对胶质瘤中多个miR异常表达及其机制的研究将对进一步探讨胶质瘤的分子病理及其诊治开拓新途径.     

Evaluation of the antitumoral effect of dihydrocucurbitacin-B in both in vitro and in vivo models

Aims  We evaluated both in vitro and in vivo antitumoral properties of an isolated compound from Wilbrandia ebracteata, dihydrocucurbitacin-B (DHCB), using B16F10 cells (murine melanoma). Materials and methods  We made use of MTT and ³H-Thymidine assays to investigate the cell viability and cell proliferation, flow cytometry analysis to monitor cell cycle and apoptosis, western blot analysis to evaluate the expression of cell cycle proteins, imunofluorescence analysis and in vivo tumor growth and metastasis. Results  Dihydrocucurbitacin-B significantly reduced cell proliferation without important effects on cells viability. DHCB lead cells to accumulate in G2/M phases accompanied by the appearance of polyploid cells, confirmed by fluorescence assays that demonstrated a remarkable alteration in the cell cytoskeleton and formation of binuclear cells. Annexin-V-FITC incorporation demonstrated that DHCB did not induce apoptosis. About 10 μg/mL DHCB was found to decrease cyclin-A, and especially in cyclin-B1. The in vivo experiments showed that DHCB treatment (once a day up to 12 days; p.o.) was able to reduce the tumor growth and lung metastasis up to 83.5 and 50.3%, respectively. Conclusions  Dihydrocucurbitacin-B reduces cell proliferation due to a decrease in the expression of cyclins, mainly cyclin-B1 and disruption of the actin cytoskeleton, arresting B16F10 cells in G2/M phase. Taken together, the in vitro and in vivo experiments suggest that DHCB was effective against cancer, however, it remains to be proved if DHCB will be a good candidate for drug development.     

中国乳腺癌患者生活质量研究进展

生活质量（qualityoflife,QOL）又译作生命质量、生存质量,它是在世界卫生组织提倡的健康新概念“人们在躯体上、精神上及社会生活中处于一种完好的状态,而不仅仅是没有患病和衰弱”的基础上构建的,是医学模式由生物医学模式向生物一心理一社会医学模式转变的体现。西方发达国家已将此概念广泛应用于临床试验、卫生政策制定和卫生资源效益评价等众多领域。生存质量已作为评价肿瘤患者术后状况的首选指标。     

Survey of changes in dietary preferences in cancer patients in China

Objective The aim of this study was to investigate the changes in dietary preferences in cancer patients in China and to determine the need for encouraging the adherence to a sensible diet among such patients.Methods A total of 468 cancer patients were interviewed using a self-designed questionnaire focusing on changes in the intake of specific foods. Data were analyzed using SPSS 16.0. Results Most patients completely avoided roosters and carp(73.1%), condiments(51.9%), and meat of aquatic species(40.4%). All other types of the specific foods were completely avoided by different subpopulations of the patients.Conclusion In addition to focusing on disease treatment, medical professionals need to help cancer patients overcome barriers associated with the customs of avoiding specific foods encompassed by the term ”fawu” and provide them with dietary guidance in order to prevent negative nutritional effects.     

新疆维吾尔族妇女宫颈病变中DAPK表达的研究

[目的]探讨DAPK基因与新疆维吾尔族妇女宫颈病变的相关关系。[方法]选取维吾尔族妇女正常或炎症的宫颈组织30例、CINⅠ30例、CINⅡ/Ⅲ30例、宫颈鳞癌组织30例采用免疫组化SP法检测DAPK蛋白表达;为了进一步验证DAPK蛋白表达水平,采用逆转录聚合酶链反应（RT-PCR）法检测正常或炎症的宫颈组织10例、CINⅠ10例、CINⅡ/Ⅲ15例、宫颈鳞癌组织20例中mRNA的表达。[结果]DAPK的蛋白表达率在正常或炎症的宫颈组织、CINⅠ、CINⅡ/Ⅲ、宫颈鳞癌组织中分别为93.3%、83.3%、60.0%、33.3%,SCC组阳性表达率明显减少（P〈0.05）;四组mRNA的表达率分别为90.0%、90.0%、46.7%、10.0%,SCC组阳性表达率明显减少（P〈0.05）。宫颈组织中DAPK在蛋白水平和mRNA水平的表达均和宫颈病变程度成负关（P〈0.05）。[结论]新疆维吾尔族妇女宫颈病变组织中DAPK蛋白和mRNA水平均随病变程度加深而减少;DAPK蛋白检测可能为宫颈癌的早期诊断提供依据。