CYB5D2对乳腺癌皮下移植瘤模型大鼠肿瘤生长的影响

[目的]研究CYB5D2对乳腺癌皮下移植瘤模型大鼠肿瘤生长的影响。[方法]通过乳腺癌MCF-7细胞系建立乳腺癌裸鼠皮下移植瘤动物模型。将含有CYB5D2表达的质粒稳定转染MCF-7细胞,多点注射,将稳定转染的细胞注入裸鼠皮下,作为观察组,同时设置shRNA-CYB5D2阴性质粒为阴性对照组,PBS为空白对照组。测量各组大鼠肿瘤体积及质量;裸鼠转移瘤模型建立成功后处死小鼠,qRT-CPR法检测肿瘤组织内CYB5D2 mRNA表达水平;Western Blot法检测各组裸鼠转移瘤组织内上皮细胞间质转化(EMT)标志表白表达情况,酶联免疫吸附法检测裸鼠转移瘤细胞上清液中MMP-2及MMP-9水平。[结果]观察组转移瘤平均体积及质量显著低于阴性组与空白组(P<0.05),阴性组与空白组裸鼠转移瘤体积及质量无明显差异(P>0.05)。提取试验小鼠肿瘤组织总RNA,qRT-CPR法检测提示观察组肿瘤组织内CYB5D2 mRNA表达量显著高于阴性组及空白对照组(P<0.05),而阴性组与空白组间差异无显著性差异(P>0.05);Western Blot法检测结果显示,观察组转移瘤组织内E-cadherin、β-catenin蛋白表达水平显著高于与阴性组及空白组(P<0.05),而N-cadherin及Snail蛋白表达显著低于与阴性组及空白组(P<0.05)。阴性组与空白组转移瘤组织内E-cadherin、β-catenin、N-cadherin及Snail蛋白表达无显著性差异(P>0.05);观察组裸鼠转移瘤细胞上清液中MMP-2及MMP-9水平显著低于与阴性组及空白组(P<0.05),而阴性组与对照组MMP-2及MMP-9水平无显著性差异(P>0.05)。[结论]上调CYB5D2基因可有效抑制乳腺癌皮下移植模型大鼠瘤体生长,推测上调CYB5D2基因可能通过下调N-cadherin、Snail、MMP2及MMP9,上调E-cadherin、β-catenin,逆转EMT,发挥抗肿瘤功效。     

维生素D_3联合5-氟尿嘧啶对人食管癌Eca-109细胞移植瘤VDR表达的影响

探讨维生素D3、5-氟尿嘧啶单独与联合使用对人食管癌Eca-109细胞移植瘤维生素D受体(vitamin Dreceptor,VDR)的作用.随机分为对照组(C)、预处理组(PT)、维生素D3组(V)、5-氟尿嘧啶组(F)、预处理+5-氟尿嘧啶组(PT+F)、维生素D3+5-氟尿嘧啶组(V+F).体外培养人食管癌Eca-109细胞,BALB/c裸鼠皮下荷瘤,2.5μg/kg1,25-(OH)2维生素D3、25 mg/kg 5-氟尿嘧啶单独与联合腹腔注射,观察瘤体生长情况,逆转录聚合酶链反应(RT-PCR)和蛋白质印迹技术(Western blot)检测裸鼠瘤体组织VDR mRNA与蛋白的表达.研究发现1,25-(OH)2维生素D3、5-氟尿嘧啶均能抑制裸鼠移植瘤的生长,PT、V、F、PT+F、V+F组瘤体体积与C组比较差异有统计学意义(P<0.05);RT-PCR与Western blot结果显示经1,25-(OH)2维生素D3单独与联合5-氟尿嘧啶使用后瘤体组织中VDR mRNA和蛋白表达升高,且联合用药更为显著(P<0.05).结果表明1,25-(OH)2维生素D3、5-氟尿嘧啶均能抑制人食管癌Eca-1...     

纳米四氧化三铁双歧杆菌LTA对实验性胃癌增殖及凋亡的影响

目的 观察纳米四氧化三铁(Fe3O4)双歧杆菌脂磷壁酸(lipoteichoicacid,LTA)对实验性胃癌增殖细胞核抗原(PCNA)和bcl-2、bax蛋白表达的影响.方法 用胃癌BGC-823细胞裸鼠皮下移植瘤模型,观察不同浓度纳米Fe3O4双歧杆菌LTA组与LTA组及环磷酰胺组移植瘤的生长情况,免疫组化法检测bcl-2和bax及增殖细胞核抗原(PCNA)的表达.结果 纳米Fe3O4双歧杆菌LTA各剂量组胃癌移植瘤PCNA阳性细胞面密度以及bcl-2阳性细胞面密度均显著低于肿瘤对照组(P<0.01),而bax 的阳性细胞面密度显著高于肿瘤阴性对照组(P<0.01).结论 纳米四氧化三铁LTA能明显降低人胃癌移植瘤的增殖活性及增强bax的蛋白表达,同时使其bcl-2基因蛋白的表达下调.     

血浆M-CSF、MMP9、TIMP-1水平及HPV阳性大鼠宫颈癌增殖能力的关系研究

摘要 目的：探究血浆巨噬细胞集落刺激因子（Macrophage colony stimulating factor, M-CSF）、基质金属蛋白酶9（Matrix metalloproteinase 9, MMP9）及其组织抑制因子1（tissue inhibitor of the metalloproteinases, TIMP1）水平及人乳头瘤病毒（Human papilloma virus,HPV）阳性大鼠宫颈癌增殖能力的关系。方法：20只健康雌性Wistar白化大鼠根据实验目的分为两组：对照组（异种移植时注射SiHa细胞作为对照实验,n=10）和观察组（将转染sh-M-CSF、sh-MMP9和sh-TIMP-1的SiHa细胞注射大鼠的子宫颈,n=10）。通过ELISA测定大鼠血浆M-CSF、MMP9和TIMP-1的水平。通过PCR检测实验大鼠中M-CSF、MMP9和TIMP-1的mRNA表达。使用数字游标卡尺分析大异种移植大鼠肿瘤体积生长。第3、4、5周分别处死并切除大鼠肿瘤进行称重。通过免疫组织化学分析肿瘤组织中增殖细胞核抗原（Proliferating cell nuclear antigen,PCNA）、pAKT和pSTAT3的蛋白表达。通过免疫组织化学染色和TUNEL染色分别确定Ki67阳性细胞数量及凋亡细胞数量。结果：观察组较对照组M-CSF、MMP9和TIMP-1的水平降低（P<0.05）。观察组较对照组M-CSF、MMP9和TIMP-1的mRNA表达降低（P<0.05）。随着时间的增加,两组大鼠肿瘤体积均增加。1周和2周对照组和观察组大鼠肿瘤体积比较无差异（P>0.05）,第3周、第4周和第5周,观察组较对照组大鼠肿瘤体积降低（P<0.05）。观察组较对照组大鼠体内肿瘤重量减少（P<0.05）。观察组较对照组PCNA、pAKT和pSTAT3的蛋白表达量降低（P<0.05）。观察组较对照组Ki67 阳性细胞数量降低,凋亡细胞升高（P<0.05）。结论：降低血浆M-CSF、MMP2和TIMP1水平可促进HPV阳性大鼠宫颈癌细胞凋亡,有效抑制细胞增殖。     

姜黄素抑制大鼠C6 胶质瘤生成及机制初步研究

目的:观察姜黄素对大鼠C6胶质瘤的抑制作用。方法:将SD大鼠随机分为假手术组、模型组、姜黄素高、低剂量组。各组动物于移植术后7天给予干预,姜黄素组给予姜黄素灌胃,14天后处死。分离肿瘤测量其体积,realtime RT-PCR法和western blot法检测肿瘤组织中核转录因子-κ B(nuclear factor-κ B,NF-κB)、EGFR mRNA和蛋白的表达。结果:姜黄素能明显降低肿瘤体积,降低肿瘤组织中NF-κ B、EGFR mRNA和蛋白的表达。结论:姜黄素能抑制大鼠C6胶质瘤的生长,其机制可能与抑制NF-κ B、EGFR表达有关。     

紫杉醇抑制前列腺癌增殖的体内实验研究

目的探讨紫杉醇对人前列腺癌细胞PC-3增殖的体内抑制作用。方法建立体内绿色荧光蛋白（GFP）标记的人雄激素非依赖性前列腺癌细胞PC-3裸鼠原位移植瘤模型,观察紫杉醇对裸鼠前列腺癌原位移植瘤的体积、重量的影响。结果裸鼠模型体内实验显示,与对照组（100μL生理盐水）相比,紫杉醇处理组（0.5 mg/kg）在给药第18天后能显著抑制前列腺肿瘤的体积（P〈0.05）;紫杉醇处理组在抑制前列腺肿瘤重量方面与对照组相比亦有明显抑制作用（P〈0.05）。与对照组相比G31P处理组VEGF（P〈0.05）的表达差异具有统计学意义（免疫组化法）。结论紫杉醇在体内实验中能明显抑制人雄激素非依赖性前列腺癌细胞系PC-3的增殖。     

姜黄素对兔骨关节炎软骨组织学及CL-2、Mmp9、COX-2、IKXβ、NF-K bmRNA表达的影响

目的:研究姜黄素对兔关节炎软骨组织的影响.方法:30只新西兰白兔,随机分成对照(A)组、姜黄素(B)组、关节炎(C)组,每组10只.除A组外,其余两组木瓜蛋白酶造骨性关节炎早期模型,后使用5%姜黄素9mg/(kg.week),生理盐水0.5ml关节腔注射治疗后,采用RT-PCR检测三组的CL-2、Mmp9、COX-2、IKKβ、NF-K b的mRNA表达情况;采用Tunel法检测软骨细胞的凋亡情况.结果:经过四周治疗,B组的CL-2的mRNA表达比C组增强(P<0.05),Mmp9、COX-2、IKKβ、NF-K b的mRNA表达明显减弱(P<0.05),而B组和C组的CL-2的mRNA表达均比A组减弱(P<0.05),在Mmp9、COX-2、IKKβ、NF-K b的mRNA表达增强(P<0.05).B组和C组凋亡指数有统计学意义(P<0.05).结论:姜黄素关节腔注射可以明显缓解关节软骨的退变,其作用可能与IL-β/IKKβ/NF-K b/COX-2、Mmp9两个正反馈信号阻断有关.     

肿瘤干细胞标记物CD_(133)与miR-429在大肠癌组织中的表达及其相关性研究

目的:检测CD133不同亚群大肠癌细胞HT-29的miR-429表达情况,探讨miR-429及CD133的表达与肿瘤的发生发展之间的关系。方法:采用荧光活化细胞分选法(FACS)分选出CD133不同亚群细胞,实时荧光定量PCR分别检测两组细胞miR-429的表达,合成miR-429寡核苷酸和阴性对照miRNA并分别转染CD133+和CD133-两个亚群细胞。再将细胞种植于非肥胖糖尿病/严重联合免疫缺陷(NOD/SCID)小鼠体内构建移植瘤模型,不同时间测量肿瘤体积和重量,RT-PCR及蛋白质印迹检测CD133+和CD133-两组肿瘤CD133mRNA和蛋白质表达。结果:血清检出CD133+细胞为67.9%,miR-429的表达量是CD133+细胞的(1.83±0.91)倍(P0.05),CD133+比例与miR-429表达呈负相关(r=0.591,P0.05);miR-429+/CD133+组的移植瘤体积及重量与对照组比较有统计学差异(P0.05),且miR-429+/CD133+组成瘤时间较对照组晚约2周,但miR-429+/CD133+组的移植瘤CD133表达量低,与阴性对照组比较无明显差异(P0.05)。结论:miR-429可能作为CD133的负性调控因子,具有抑制肿瘤生长的作用,但miR-429与CD133在肿瘤发生、发展过程中的作用机制有待进一步研究阐明。     

紫杉醇联合姜黄素对人前列腺癌PC3细胞侵袭和老化的影响

目的:选用姜黄素与紫杉醇联合作用于前列腺癌PC3细胞株,从细胞水平观察姜黄素和紫杉醇合用对癌细胞增殖、侵袭和老化的影响,以进一步寻找综合治疗前列腺癌的有效措施。方法:低剂量姜黄素和紫杉醇作用于PC3细胞,用CCK-8、β-半乳糖苷染色、划痕侵袭实验等方法从细胞增殖、侵袭和老化等方面观察对PC3的影响。结果:姜黄素联合紫杉醇可协同降低PC3细胞增殖活性(P<0.01);划痕实验显示,姜黄素和紫杉醇合用可降低细胞的侵袭浸润能力;低浓度的姜黄素还可促使PC3细胞老化。结论:在细胞水平,紫杉醇联合姜黄素可协同降低PC3细胞增殖分化活性,减弱肿瘤细胞的侵袭浸润能力。     

高氧对雄激素敏感的前列腺癌细胞移植瘤生长及其缺氧诱导因子-1α表达的影响

目的:探讨高氧对雄激素敏感的前列腺癌细胞移植瘤生长及其缺氧诱导因子-1α表达的影响。方法:将前列腺癌前列腺淋巴结癌(LNCa P)细胞接种于36只Foxn1小鼠的双侧腹,并将其随机放置于含氧量不同的气室中并分组如下:缺氧组11例,常氧组16例,高氧组9例。处理28天后进行称重,麻醉处死,从左心室取血样;分离出移植瘤并进行称重。采用Western blotting、免疫荧光分析、血红蛋白测定的方法对各组移植瘤生长、血管生成及血管化、缺氧诱导因子-1(HIF-1α)表达以及细胞信号转导因子表达进行检测。结果:缺氧组的移植瘤生长较常氧组快(P0.05);高氧组移植瘤生长与常氧组相比差异不具有统计学意义(P0.05)。高氧组移植瘤的HIF-1α、血管内皮生长因子(VEGF)和血管内皮生长因子受体2(VEGF-R2)表达均较常氧组高,而缺氧组移植瘤的HIF-1α表达与常氧组基本相似。缺氧组移植瘤的血[HB]增长率(175%)高于常氧组(45%)。高氧组的Nrf2的表达水平较常氧组明显增加(P0.05)。结论:体内高氧诱导HIF-1α在LNCa P肿瘤高表达的同时,不会加快肿瘤的生长。     

Efficient Drug Delivery of Paclitaxel Glycoside: A Novel Solubility Gradient Encapsulation into Liposomes Coupled with Immunoliposomes Preparation

Although the encapsulation of paclitaxel into liposomes has been extensively studied, its significant hydrophobic and uncharged character has generated substantial difficulties concerning its efficient encapsulation into the inner water core of liposomes. We found that a more hydrophilic paclitaxel molecule, 7-glucosyloxyacetylpaclitaxel, retained tubulin polymerization stabilization activity. The hydrophilic nature of 7-glucosyloxyacetylpaclitaxel allowed its efficient encapsulation into the inner water core of liposomes, which was successfully accomplished using a remote loading method with a solubility gradient between 40% ethylene glycol and Cremophor EL/ethanol in PBS. Trastuzumab was then conjugated onto the surface of liposomes as immunoliposomes to selectively target human epidermal growth factor receptor-2 (HER2)-overexpressing cancer cells. In vitro cytotoxicity assays revealed that the immunoliposomes enhanced the toxicity of 7-glucosyloxyacetylpaclitaxel in HER2-overexpressing cancer cells and showed more rapid suppression of cell growth. The immunoliposomes strongly inhibited the tumor growth of HT-29 cells xenografted in nude mice. Notably, mice survived when treated with the immunoliposomes formulation, even when administered at a lethal dose of 7-glucosyloxyacetylpaclitaxel in vivo. This data successfully demonstrates immunoliposomes as a promising candidate for the efficient delivery of paclitaxel glycoside.     

藏红花素对人膀胱移行细胞癌T24细胞裸鼠移植瘤的实验研究

目的检测藏红花素（crocin）对人膀胱移行细胞癌T24细胞株裸鼠移植瘤的作用。方法12只裸鼠右后腿背侧皮下接种T24移植瘤细胞后随机分为PBS对照组（6只）和50mmol/L藏红花素治疗组（6只）。从接种后第28d开始,每3d一次于肿瘤局部注射给予PBS或藏红花素,每3d测量1次肿瘤大小,并绘制移植瘤生长曲线;处理后第21d杀鼠摘取移植瘤,光镜、电镜观察肿瘤组织形态改变;RT-PCR检测Bcl-2、Bax mRNA表达;SP-免疫组织化学法分析sur-vivin、cyclinD1蛋白表达。结果藏红花素对人膀胱癌T24细胞裸鼠移植瘤的生长具有抑制作用,抑制率达28.7%。HE染色结果显示肿瘤标本内出现片状坏死灶;电镜观察可见凋亡的形态学改变;RT-PCR检测结果显示Bcl-2的表达水平下降,而Bax的表达水平上调;免疫组化结果显示经藏红花素处理后survivin、CyclinD1的表达下调。结论藏红花素能够抑制人膀胱癌T24细胞裸鼠移植瘤的生长,作用的分子机制与下调Bcl-2、survivin、cyclinD1和上调Bax基因表达有关。     

Synergistic effect of curcumin on epigallocatechin gallate-induced anticancer action in PC3 prostate cancer cells

Epigallocatechin gallate (EGCG) and curcumin are well known to naturally-occurring anticancer agents. The aim of this study was to verify the combined beneficial anticancer effects of curcumin and EGCG on PC3 prostate cancer cells, which are resistant to chemotherapy drugs and apoptosis inducers. EGCG showed weaker inhibitory effect on PC3 cell proliferation than two other prostate cancer cell lines, LNCaP and DU145. Co-treatment of curcumin improved antiproliferative effect of EGCG on PC3 cells. The protein expressions of p21 were significantly increased by the co-treatment of EGCG and curcumin, whereas it was not changed by the treatment with each individual compound. Moreover, treatments of EGCG and curcumin arrested both S and G2/M phases of PC3 cells. These results suggest that the enhanced inhibitory effect of EGCG on PC3 cell proliferation by curcumin was mediated by the synergic up-regulation of p21-induced growth arrest and followed cell growth arrest. [BMB Reports 2015; 48(8): 461-466]     

替吉奥联合三维适形放疗对胰腺癌细胞生物学的影响

目的：探究替吉奥复方制剂联合三维适形放射疗法对胰腺癌细胞生物学的影响及安全性评价。方法：培养PANC-1人胰腺癌细胞,构建PANC。1胰腺癌裸鼠异位肿瘤模型并随机进行分组。观察替吉奥联合三维适形放疗对胰腺癌肿瘤细胞生长的抑制作用,记录荷瘤裸鼠的肿瘤体积以及生存期。同时观察联合治疗所产生的副作用。结果：相比于单独用药组和单独放疗组,联合组能够显著抑制肿瘤体积生长,延长荷瘤裸鼠的中位生存期。结论：替吉奥单独给药和单独三维适形放疗均能抑制肿瘤的生长,二者联合使用能够发挥协同作用,是一种潜在的高效的胰腺癌治疗手段。     

Effectiveness of the Histone Deacetylase Inhibitor (S)-2 against LNCaP and PC3 Human Prostate Cancer Cells

Histone deacetylase inhibitors (HDACi) represent a promising class of epigenetic agents with anticancer properties. Here, we report that (S)-2, a novel hydroxamate-based HDACi, shown previously to be effective against acute myeloid leukemia cells, was also a potent inducer of apoptosis/differentiation in human prostate LNCaP and PC3 cancer cells. In LNCaP cells (S)-2 was capable of triggering H3/H4 histone acetylation, H2AX phosphorylation as a marker of DNA damage and producing G₀/G₁ cell cycle arrest. Consistently, (S)-2 led to enhanced expression of both the protein and mRNA p21 levels in LNCaP cells but, contrary to SAHA, not in normal non-tumorigenic prostate PNT1A cells. Mechanistic studies demonstrated that (S)-2-induced apoptosis in LNCaP cells developed through the cleavage of pro-caspase 9 and 3 and of poly(ADP-ribose)-polymerase accompanied by the dose-dependent loss of mitochondrial membrane potential. Indeed, the addition of the pan-caspase inhibitor Z-VAD-fmk greatly reduced drug-mediated apoptosis while the antioxidant N-acetyl-cysteine was virtually ineffective. Importantly, preliminary data with nude mice xenografted with LNCaP cells showed that (S)-2 prompted a decrease in the tumor volume and an increase in H2AX phosphorylation within the cancer cells. Moreover, the highly metastatic prostate cancer PC3 cells were also sensitive to (S)-2 that: i) induced growth arrest and moderate apoptosis; ii) steered cells towards differentiation and neutral lipid accumulation; iii) reduced cell invasiveness potential by decreasing the amount of MMP-9 activity and up-regulating TIMP-1 expression; and iv) inhibited cell motility and migration through the Matrigel. Overall, (S)-2 has proven to be a powerful HDACi capable of inducing growth arrest, cell death and/or differentiation of LNCaP and PC3 prostate cancer cells and, due to its low toxicity and efficacy in vivo, might also be of clinical interest to support conventional prostate cancer therapy.     

Growth of tumor cells with different sensitivities for murine natural killer cells in young and adult athymic nude mice

Of four tumor cell lines, the murine YAC lymphoma, the human K562 lymphoma, and the human prostatic carcinomas PC3 and PC93, the susceptibility to murine natural killer (NK) cells as well as the tumorigenicity in young (3.5-4 weeks old) and in adult (8-10 weeks old) nude mice were studied. In young nude mice, which exhibited a lower level of NK cell activity than adult nude mice, the formation of solid tumors after inoculation of tumor cell suspensions occurred more frequently and with a shorter time lag than in adult animals. These effects were observed not only with the NK-sensitive YAC cells, but also with the relatively NK-insensitive PC3 and PC93 cells, indicating that also factors other than NK cell susceptibility may influence the growth of tumor cells in nude mice. Therefore, the use of young nude mice may enhance the rate of success of heterotransplantation of human tumors, regardless of the NK cell susceptibility of the tumor cells.     

Cancer-Associated Fibroblasts in a Human HEp-2 Established Laryngeal Xenografted Tumor Are Not Derived from Cancer Cells through Epithelial-Mesenchymal Transition,Phenotypically Activated but Karyotypically Normal

Cancer-associated fibroblasts (CAFs) play a crucial role in cancer progression and even initiation. However, the origins of CAFs in various cancer types remain controversial, and one of the important hypothesized origins is through epithelial-mesenchymal transition (EMT) from cancer cells. In this study, we investigated whether the HEp-2 laryngeal cancer cells are able to generate CAFs via EMT during tumor formation, which is now still unknown. The laryngeal xenografted tumor model was established by inoculating the HEp-2 laryngeal cancer cell line in nude mice. Primary cultured CAFs from the tumor nodules and matched normal fibroblasts (NFs) from the adjacent connective tissues were subcultured, purified, and verified by immunofluorescence. Migration, invasion, and proliferation potentials were compared between the CAFs and NFs. A co-culture of CAFs with HEp-2 cells and a co-injection of CAFs with HEp-2 cells in nude mice were performed to examine the cancer-promoting potential of CAFs to further verify their identity. Karyotypic analyses of the CAFs, NFs, and HEp-2 cells were conducted. A co-culture of NFs with HEp-2 cells was also performed to examine the expression of activated markers of CAFs. A pathological examination confirmed that the laryngeal xenografted tumor model was successfully established, containing abundant CAFs. Immunocytochemical staining verified the purities and identities of the CAFs and NFs. Although the CAFs manifested higher migration, invasion, proliferation, and cancer-promoting capacities compared with the NFs, an analysis of chromosomes revealed that both the CAFs and NFs showed typical normal mouse karyotypes. In addition, the NFs co-cultured with HEp-2 cells did not show induced expressions of activated markers of CAFs. Our findings reveal that the CAFs in the HEp-2 established laryngeal xenografted tumor are not of laryngeal cancer origin but of mouse origin, indicating that the HEp-2 laryngeal cancer cells cannot generate their own CAFs via EMT in this model.     

Opposite angiogenic outcome of curcumin against ischemia and Lewis lung cancer models: in silico,in vitro and in vivo studies

The aim of this study was to investigate the angiogenic effects of curcumin on an ischemia and lung cancer model. To induce ischemia combined with lung cancer models, unilateral femoral arteries of C57BL/6 mice were disconnected on one side of the mouse and Lewis lung carcinoma (LLC) cells were xenografted on the opposite side. Angiogenic effects and underlying mechanisms associated with curcumin were investigated. Molecular target(s), signaling cascades and binding affinities were detected by Western blot, two-dimensional gel electrophoresis (2-DE), computer simulations and surface plasmon resonance (SPR) techniques. Curcumin promoted post-ischemic blood recirculation and suppressed lung cancer progression in inbred C57BL/6 mice via regulation of the HIF1α/mTOR/VEGF/VEGFR cascade oppositely. Inflammatory stimulation induced by neutrophil elastase (NE) promoted angiogenesis in lung cancer tissues, but these changes were reversed by curcumin through directly reducing NE secretion and stimulating α1-antitrypsin (α1-AT) and insulin receptor substrate-1 (IRS-1) production. Meanwhile, curcumin dose-dependently influenced endothelial cells (EC) tube formation and chicken embryo chorioallantoic membrane (CAM) neovascularization. Curcumin had opposite effects on blood vessel regeneration under physiological and pathological angiogenesis, which was effected through negative or positive regulation of the HIF1α/mTOR/VEGF/VEGFR cascade. Curcumin had the promise as a new treatment modality for both ischemic conditions and lung cancer simultaneously in the clinic.     

Differential protein expressions induced by adenovirus-mediated p16 gene transfer into Balb/c nude mouse

To evaluate the safety of adenovirus-mediated gene transfer, we investigated differential protein expression after transducing adenoviral vector containing the p16(INK4a) tumor suppressor gene (Ad5CMV-p16) into Balb/c nude mice. We found that adenovirus-mediated p16(INK4a) gene transfer inhibited experimental lung metastasis, and that the intratumoral injection of Ad5CMV-p16 resulted in regression of A549 cell xenografted tumors in Balb/c nude mice. We investigated changes in protein expression after intratumoral injection of Ad5CMV-p16 or Ad5CMV (10(10) plaque-forming units) into A549 cell xenografted Balb/c nude mice by two-dimensional gel electrophoresis /matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Compared with the control (serum-free medium treated tumor cells) Ad5CMV-p16 gene transfer changed the expression of 29 proteins including heterogeneous nuclear ribonucleoprotein, protein phosphatase 2, 14-3-3 zeta protein, alpha-tubulin, and glutathione-S-transferase P1. Moreover, both Ad5CMV-p16 and Ad5CMV up-regulated the expression of glutathione-S-transferase P1. In addition, Ad5CMV-p16 gene transfer did not seem to increase the expression of tumorigenicity-related protein in Balb/c nude mice. Further studies will be needed to investigate the effect of Ad5CMV-p16 on normal human cells and tissues for safety evaluation. These results suggest that the p16 gene seems to have an important role in apoptosis as well as in cell cycle arrest in non-small cell lung cancer.     

Establishment of a Chinese bladder cancer cell line (T921) with high metastatic activity

Muscle-invasive bladder cancer is prone to metastasis without a standard organ preference. The current cell lines used to study bladder cancer have primarily been derived from individuals in Western populations, and no human bladder cancer cell line has been established from the Chinese population. A bladder cancer cell line was derived from a female Chinese patient with muscle-invasive bladder cancer, and these cells were then xenografted into the bladders of three nude mice. Five weeks later, these mice were killed to observe local invasion and distant metastasis. The metastatic tumors were also removed and analyzed to assess the metastatic mechanism. This bladder cancer cell line, named T921, was successfully established, as evidenced by karyotype and immunohistochemistry analyses. Multi-organ metastases were observed in all three of the nude mice 5 wk after the orthotopic transfer of the cell line. In addition, epithelial–mesenchymal transition (EMT)-related genes were involved in the tumor metastases. The T921 bladder cancer cell line was successfully established, and EMT was observed to play a role in bladder cancer metastasis.