Translational evidence on the role of Src kinase and activated Src kinase in invasive breast cancer

Src kinase is a member of a non-receptor tyrosine kinase family. It has been implicated as a regulator of cell proliferation and survival and plays a complex role in cell adhesion and motility. In vitro evidence for a role for Src in breast cancer is compelling. However, only a few translational clinical studies have been undertaken in this field. This review summarises translational evidence on expression and activation of Src kinase in breast cancer patient cohorts exploring clinical significance and the possibility of identifying key biomarkers. There is strengthened translational proof for a definitive role of Src in breast cancer. Nevertheless, there remains a need to find a robust biomarker to identify patients responsive to Src inhibitors for clinical trials.     

Src激酶在胃癌细胞来源的exosome促进肿瘤细胞增殖中的作用

目的:研究胃癌细胞来源的外泌体(exosome)对肿瘤细胞增殖的影响,初步探讨Src蛋白激酶在此过程中的作用.方法:采用离心超滤和蔗糖密度梯度超速离心的方法从胃癌SGC7901细胞的上清液中分离出胃癌细胞来源的exosome.透射电子显微镜下观察exosome形态.MTT法检测细胞增殖能力,Western blot检测...     

新的血管生成抑制肽(手性制剂)抗肿瘤血管生成的实验研究

目的 观察一种新的血管生成抑制胜对肿瘤的抑制作用，并探讨其作用机制。方法 体外药效实验，采用MTT法及血管内皮细胞迁移法，体内用Lewis肺癌瘤株皮下接种C57BL／6N小鼠，观察皮下移植瘤生长情况。结果 新的血管生成抑制胜对血管内皮细胞增殖、迁移有明显的抑制作用，在体内使Lewis肺癌皮下移植田体积明显缩小，毛细血管稀少。结论 新的血管生成抑制肽能明显抑制Lewis的肿瘤生长，其机制可能与抑制肿瘤血管生成有关。     

Zinc ligand-disrupted recombinant human Endostatin: Potent inhibition of tumor growth, safety and pharmacokinetic profile

Endostatin, a potent endogenous inhibitor of angiogenesis, inhibits the growth of primary tumors without induction of acquired drug resistance in mice. We report that a soluble recombinant human (rh) Endostatin produced with characteristics of the native Endostatin, effectively inhibited the growth of primary tumors and pulmonary metastases in a dose-dependent manner. We also show that deletion of two of the four zinc ligands of rhEndostatin did not affect this potent tumor inhibiton. The growth of established Lewis lung primary tumors implanted into mice was inhibited (80–90%) upon systemic treatment with 50 mg/kg/12 h of rhEndostatin. Using the B16-BL6 murine experimental pulmonary metastases model, rhEndostatin administered at 1.5 mg/kg/day or 4.5 mg/kg/day beginning 3- or 11-days post tumor cell injection, respectively, resulted in an approximate 80% inhibition of tumor growth. At effective anti-tumor doses of 1.5 and 50 mg/kg, pharmacokinetic modeling in mice showed (a) the protein was 100% bioavailable, (b) the AUC ranged from 16 to 700 ngml/h and (c) the C_max ranged from 161 to 4582 ng/ml. At the highest dose tested (300 mg/kg), delivered as a single bolus, no drug-related toxicity was observed in a Cynomolgus monkey infused with rhEndostatin. No toxicity was observed even at AUC and C_max values that were 1.3- to 56-fold higher than those observed in mice with tumors that were potently inhibited. Our production system yields a well characterized, soluble and potent rhEndostatin at quantities sufficient for human use. The preclinical studies described herein are an important first step toward the assessment of Endostatin in the clinic. This revised version was published online in June 2006 with corrections to the Cover Date.     

Aminoguanidine impedes human pancreatic tumor growth and metastasis development in nude mice

AIM： To study the action of aminoguanidine on pancreatic cancer xenografts in relation to cell proliferation, apoptosis, redox status and vascularization. METHODS： Xenografts of PANC-1 cells were developed in nude mice. The animals were separated into two groups： control and aminoguanidine treated. Tumor growth, survival and appearance of metastases were determined in vivo in both groups. Tumors were excised and ex vivo histochemical studies were performed. Cell growth was assessed by Ki-67 expression. Apoptosis was studied by intratumoral expression of B cell lymphoma-2 protein （Bcl-2） family proteins and Terminal deoxynucleotidyl transferase biotin-dUTP Nick End Labeling （Tunel）. Redox status was evaluated by the expression of endothelial nitric oxide synthase （eNOS）, catalase, copper-zinc superoxide dismutase （CuZnSOD）,manganese superoxide dismutase （MnSOD） and glutathione peroxidase （GPx）. Finally, vascularization was determined by Massons trichromic staining, and by VEGF and CD34 expression. RESULTS： Tumor volumes after 32 d of treatment by aminoguanidine （AG） were significantly lower than in control mice （P 〈 0.01）. Median survival of AG mice was significantly greater than control animals （P 〈 0.01）. The appearance of both homolateral and contralateral palpable metastases was significantly delayed in AG group. Apoptotic cells, intratumoral vascularization （trichromic stain） and the expression of Ki-67, Bax, eNOS, CD34, VEGF, catalase, CuZnSOD and MnSOD were diminished in AG treated mice （P 〈 0.01）, while the expression of Bcl-2 and GPx did not change. CONCLUSION： The antitumoral action of aminoguanidine is associated with decreased cell proliferation, reduced angiogenesis, and reduced expression of antioxidant enzymes.     

Clinicopathological correlation of radiologic measurement of post-therapy tumor size and tumor volume for pancreatic ductal adenocarcinoma

ObjectivesTumor size measurement is critical for accurate tumor staging in patients with pancreatic ductal adenocarcinoma (PDAC). However, accurate tumor size measurement is challenging in patients who received neoadjuvant therapy before resection, due to treatment-induced fibrosis and tumor invasion beyond the grossly identified tumor area. In this study, we evaluated the correlation between the tumor size and tumor volume measured on post-therapy computed tomography (CT) scans and the pathological measurement. Also, we investigated the correlation between these measurements and clinicopathological parameters and survival.Materials and methodsRetrospectively, we evaluated 343 patients with PDAC who received neoadjuvant therapy, followed by pancreaticoduodenectomy and had pre-operative pancreatic protocol CT imaging. We measured the longest tumor diameter (RadL) and the radiological tumor volume (RadV) on the post-therapy CT scan, then we categorized RadL into four radiologic tumor stages (RTS) based on the current AJCC staging (8th edition) protocol and RadV based on the median. Pearson correlation or Spearman’s coefficient (δ), T-test and ANOVA was used to test the correlation between the radiological and pathological measurement. Chi-square analysis was used to test the correlation with the tumor pathological response, lymph-node metastasis and margin status and Kaplan-Meier and Cox-proportional hazard for survival analysis. P-value < 0.05 was considered significant.ResultsAs a continuous variable, RadL showed a positive linear correlation with the post-therapy pathologic tumor size in the overall patient population (Pearson correlation coefficient: 0.72, P < 0.001) and RadV (δ: 0.63, p < 0.0001). However, there was no correlation between RadL and pathologic tumor size in patients with ypT0 and those with pathologic tumor size of ≤1.0 cm. Post-therapy RTS and RadV group correlated with ypT stage, tumor response grades using either CAP or MDA grading system, distance of superior mesenteric artery margin and tumor recurrence/metastasis.ConclusionAlthough RadL tends to understage ypT in PDAC patients who had no radiologically detectable tumor or small tumors (RTS0 or RTS1), radiologic measurement of post-therapy tumor size may be used as a marker for the pathologic tumor staging and tumor response to neoadjuvant therapy.     

胰腺癌肿标志物单项和联合检测的价值

目的 评价血清肿瘤标志物单项检测和联合检测对胰腺癌诊断的意义。方法 对10例慢性胰腺炎和51例胰腺癌病人用放免法测定血清CA199,CA50,CA125和CEA含量,分析并比较单项检测和联合检测的意义。结果 4种肿瘤标志物单项检测以CA199对胰腺癌诊断意义最大,敏感性和特异性最好,分别达92．5％和81．81％,误检率18．18％,漏检率7．5％,阳性预测值94．87％,阴性预测值75％,研究还发现CA199特别是胰体癌诊断有较大意义;联合检测以CA199＋x(CA50,CA125,CEA任一阳性）意义较大,敏感性87．5％,特异性81．81％,误检率18．18％,漏检率12．5％,阳性预测值94．59％,阴性预测值64．28％;但诊断 价值不大于CA199单项检测（P＞0．05）。结论 CA199检测对胰腺癌诊断意义较大,特别是对早期胰体癌诊断有较大意义;联合检测并不能提高CA199的诊断价值,却增加医疗费用,意义不大。     

胰腺癌患者血清十二种肿瘤标记物的变化

目的 探讨定量测定胰腺癌患者血清12种肿瘤标志物的临床意义。方法 用蛋白芯片技术定量测定42例正常人、30例胰腺癌和16例胰腺炎患者血清12种肿瘤标志物的变化并对检测效果进行评价。结果 42例正常人血清CA19-9(KU/L)、NSE(μg/L)、CEA(μg/L)、CA242(KU/L)、Ferritin(μg/L)、β-HCG(μg/L)、AFP(μg/L)、f-PSA(μg/L)、PSA(μg/L)、CA125(KU/L)、HGH(μg/L)和CA153(KU/L)的含量分别为:12.42±10.62、2.24±1.41、1.78±1.04、5.73±5.67、91.17±78.47、0.64±0.33、2.96±3.89、0.13±0.11、0.62±1.38、5.46±9.53、1.63±2.37、9.83±9.40;30例胰腺癌患者血清12种肿瘤标志物含量依次为:279.47±402.0、9.50±11.30、8.57±21.95、98.23±89.33、301.98±216.63、1.49±2.28、5.50±7.69、0.27±0.62、1,62±3.23、167.16±252.49、3.38±8.38、43.22±99.34;16例胰腺炎患者其含量依次为:53.38±86.38、12.78±28.50、1.20±0.94、22.99±51.13、343.43±308.27、3.63±11.99、5.09±3.66、0.10±1.32、0.66±0.61、98.67±212.23、1.79±1.63、4.15±2.13。该蛋白芯片测定的敏感性为70.0％,特异性为89.7％,阳性预测值为77.8％,阴性预测值为88.1％。结论 采用蛋白芯片技术同时测定患者血清中多种肿     

雷公藤内酯醇抑制胰腺癌细胞生长及血管生成的实验研究

目的 探讨雷公藤内酯醇(TL)对胰腺癌细胞生长及肿瘤新生血管生成的抑制作用.方法 采用CCK-8试剂盒检测TL对胰腺癌SW1990细胞的生长抑制作用;采用TUNEL法检测TL对细胞凋亡的诱导作用;观察尾静脉注射TL对裸鼠移植瘤生长抑制作用和对瘤组织VEGF表达、微血管密度(MVD)的影响.结果 TL呈浓度和时间依赖性抑制SW1990细胞增殖,160 mg/ml的TL干预SW1990细胞24 h,细胞抑制率达50.6%,细胞凋亡率从对照组的9.6%升高到45.1%(P<0.01);TL0.5 mg/kg体重瘤内注射对移植瘤的抑瘤率达89.9%,瘤组织VEGF表达减少,MVD从36.25±8.64减少到9.87±3.34(P<0.01).结论 TL可显著抑制胰腺癌细胞增殖并诱导细胞凋亡;通过抑制肿瘤新生血管生成可抑制裸鼠SW1990细胞移植瘤的生长.     

Effect of SU5416 on the inhibition of growth and metastasis of human gastric cancer implanted in nude mice

OBJECTIVE: To study the effects of the angio­genesis inhibitor SU5416 on the growth and liver metastasis of gastric cancer and to investigate its effects on the apoptosis of gastric cancer cells. METHODS: A model simulating the metastasis of human gastric cancer was established by orthotopic implantation of histologically intact human tumor tissue into the gastric wall of nude mice, which were randomly divided into four groups: control group (saline solution), 5‐FU group (fluorouracil 30 mg/kg per day i.p), SU5416 group (SU5416 15 mg/kg per day i.p.), and combined treatment with 5‐Fu and SU5416. Eight weeks after implantation, the tumor weight, inhibition rate, intratumoral microvessel density (MVD), apoptotic index (AI), and the status of metastasis were evaluated after the mice were killed. RESULTS: Compared with the control group, the growth of the orthotopically‐implanted tumors was significantly inhibited, with reduced weight, and the tumor inhibition rate was 44.5%, 79.3%, and 84.4%, respectively, in the mice treated with 5‐FU, SU5416 and both. The incidence of liver metastasis was also significantly decreased in the 5‐Fu, SU5416, and combination groups compared with the control group (36.4%, 25.0%, and 0% vs 90.0%). The MVD of the 5‐FU group, the SU5416 group and the combined group was 13.1 ± 4.7, 3.9 ± 1.8 and 2.1 ± 1.5, respectively, which was decreased significantly compared with the control group (14.6 ± 5.8). The AI was increased significantly in the treated mice (6.81 ± 4.92%, 9.82 ± 3.76% and 17.65 ± 9.85% vs 3.76 ± 2.25%). The growth of and liver metastasis of the human gastric cancer implanted in the nude mice were both more significantly inhibited in the SU5416 group and the combined group than in the control and 5‐FU groups (P < 0.05). CONCLUSIONS: SU5416 can induce apoptosis in gastric cancer by inhibiting tumor angiogenesis and has a strong inhibitory effect on both the growth and liver metastasis of gastric cancer implanted in nude mice. The combination of SU5416 with other cytotoxic agents is more effective.     

Circulating tumor cells in pancreatic cancer patients: Enrichment and cultivation

AIM:To investigate the feasibility of separation and cultivation of circulating tumor cells(CTCs)in pancreatic cancer(Pa C)using a filtration device.METHODS:In total,24 Pa C patients who were candidates for surgical treatment were enrolled into the study.Peripheral blood samples were collected before an indicated surgery.For each patient,approximately8 m L of venous blood was drawn from the antecubitalveins.A new size-based separation Meta Cell?technology was used for enrichment and cultivation of CTCs in vitro.(Separated CTCs were cultured on a membrane in FBS enriched RPMI media and observed by inverted microscope.The cultured cells were analyzed by means of histochemistry and immunohistochemistry using the specific antibodies to identify the cell origin.RESULTS:CTCs were detected in 16 patients(66.7%)of the 24 evaluable patients.The CTC positivity did not reflect the disease stage,tumor size,or lymph node involvement.The same percentage of CTC positivity was observed in the metastatic and non-metastatic patients(66.7%vs 66.7%).We report a successful isolation of CTCs in Pa C patients capturing proliferating cells.The cells were captured by a capillary action driven size-based filtration approach that enabled cells cultures from the viable CTCs to be unaffected by any antibodies or lysing solutions.The captured cancer cells displayed plasticity which enabled some cells to invade the separating membrane.Further,the cancer cells in the“bottom fraction”,may represent a more invasive CTC-fraction.The CTCs were cultured in vitro for further downstream applications.CONCLUSION:The presented size-based filtration method enables culture of CTCs in vitro for possible downstream applications.     

Uncoupling protein 2 deficiency of non-cancerous tissues inhibits the progression of pancreatic cancer in mice

Background: Pancreatic ductal adenocarcinoma(PDAC) is a disease of the elderly mostly because its development from preneoplastic lesions depends on the accumulation of gene mutations and epigenetic alterations over time. How aging of non-cancerous tissues of the host affects tumor progression, however, remains largely unknown. Methods: We took advantage of a model of accelerated aging, uncoupling protein 2-deficient( Ucp2 knockout, Ucp2 KO) mice, to investigate the growth of orthotopically trans...     

蛋白质组学在胰腺癌中的研究进展

胰腺癌是一种常见的恶性肿瘤,迫切需要早期诊断及治疗。快速发展的蛋白质组学技术为探讨胰腺癌发生的分子机制、发现新的肿瘤标记物及治疗靶点提供了新的手段,在胰腺癌研究中显示出巨大的前景。     

胰腺癌及其转移灶中细胞凋亡的检测及意义

为探讨细胞凋亡在胰腺癌及其转移不同阶段的作用 ,采用原位 DNA断裂点的 3-羟基末端标记(TUNEL)技术原位观察 5 0例胰腺癌患者的癌组织和 15例胰腺癌患者转移灶组织的细胞凋亡情况 ,并与 10例慢性胰腺炎组织 (对照组 )作比较。结果发现 ,细胞凋亡普遍存在于胰腺病变组织中 ,从胰腺良性病变、胰腺癌到胰腺癌转移灶 ,细胞凋亡的发生率 (AI)呈升高趋势 ,P<0 .0 5。随着病情逐渐加重 ,AI亦呈升高的趋势 ;未转移和高分化胰腺癌的 AI较有转移和低分化者低 ,但无显著性差异。认为细胞凋亡的变化规律与胰腺癌的发生、发展和临床病理特征有密切关系     

Induction of M2-macrophages by tumour cells and tumour growth promotion by M2-macrophages: A quid pro quo in pancreatic cancer

IntroductionMore effective therapies are required to improve survival of pancreatic cancer. Possible immunologic targets include tumour associated macrophages (TAMs), generally consisting of M1- and M2-macrophages. We have analysed the impact of TAMS on pancreatic cancer in a syngeneic orthotopic murine model.Methods6606PDA murine pancreatic cancer cells were orthotopically injected into C57BL6 mice. Tumour growth was monitored using MRI. Macrophages were depleted by clodronate liposomes. Tumours including microvessel density were evaluated using immunohistochemistry, immunofluorescence and/or cytometric beads assays. Naïve macrophages were generated employing peritoneal macrophages. In vitro experiments included culturing of macrophages in tumour supernatants as well as tumour cells cultured in macrophage supernatants using arginase as well as Griess assays.ResultsClodronate treatment depleted macrophages by 80% in livers (p = 0.0051) and by 60% in pancreatic tumours (p = 0.0169). MRI revealed tumour growth inhibition from 221.8 mm³ to 92.3 mm³ (p = 0.0216). Micro vessel densities were decreased by 44% (p = 0.0315). Yet, MCP-1-, IL-4- and IL-10-levels within pancreatic tumours were unchanged. 6606PDA culture supernatants led to a shift from naïve macrophages towards an M2-phenotype after a 36 h treatment (p < 0.0001), reducing M1-macrophages at the same time (p < 0.037). In vivo, M2-macrophages represented 85% of all TAMs (p < 0.0001). Finally, culture supernatants of M2-macrophages induced tumour growth in vitro by 63.2% (p = 0.0034).ConclusionsThis quid pro quo of tumour cells and M2-macrophages could serve as a new target for future immunotherapies that interrupt tumour promoting activities of TAMs and change the iNOS-arginase balance towards their tumoricidal capacities.     

花姜酮对胰腺癌细胞迁移和侵袭的影响及机制的研究

目的:探讨花姜酮(zerumbone)对人胰腺癌细胞株BxPC-3和Panc-1迁移和侵袭的影响及其相关基因的表达。方法:以BxPC-3和Panc-1为研究对象,通过四甲基偶氮唑盐(MTT)实验选择毒性较小的花姜酮浓度,用细胞划痕实验、Transwell小室迁移和侵袭实验检测药物对细胞迁移和侵袭能力的影响,蛋白质印迹(Western blot)检测各细胞组CXC族趋化因子受体4(CXCR4)、促分裂原活化蛋白酶激酶1/2(MEK1/2)、磷酸化MEK 1/2(p-MEK1/2)、细胞外调节激酶1/2(ERK1/2)、磷酸化ERK1/2(p-ERK1/2)蛋白表达变化。结果:花姜酮对BxPC-3和Panc-1细胞的生长均有抑制作用,且随着药物浓度的增高和作用时间的延长而增强(P<0.05)。低浓度药物作用后BxPC-3和Panc-1的划痕迁移率都低于对照组(P<0.05)。与对照组相比,Transwell迁移和侵袭实验中BxPC-3和Panc-1用药组的平均穿膜细胞数均减少(P     

多种肿瘤标志物在胰腺癌临床分期中的应用

目的探讨多种肿瘤标志物检测对胰腺癌的临床分期及术前评估的应用。方法45例胰腺癌,其中Ⅰ-Ⅱ期16例、Ⅲ期11例、Ⅳ期18例,检测其术前CA199、CA125、CEA等肿瘤指标,分析多种肿瘤标志物在胰腺癌临床分期及术前评估中的意义。结果不同分期胰腺癌之间CA199水平差异无显著性（P=0．381）,CA125、CEA的水平随着分期的递增而升高,但统计分析显示只有CA125组间差异有统计学意义（P〈0．05）。按照是否可切除分为两组,CA199水平在两组间差异无统计学意义（Z=1．045,P=0．296）,而CA125、CEA水平在两组间存在显著性差异（P=0．000、0．045）。多因素分析结果显示,胰腺癌的不同分期与有无黄疸、胆道疾病史、性别、体重减轻等之间存在显著统计学差异（P〈0．05）,而与术前胆红素水平、血糖、糖尿病史、年龄等之间无统计学差异。结论CA125和CEA可辅助用于胰腺癌患者临床分期及术前评估,具有无创性、病人依从性好等优点,具有较好的临床潜在应用价值。     

Tumor angiogenesis as a prognostic predictor in pancreatic cancer

The purpose of this study was to evaluate the role of angiogenesis, proliferative activity (assessed by Ki-67 expression), p53 and ras-oncogene (H-ras) expression, and conventional clinicopathologic factors in predicting overall survival rates in patients with pancreatic ductal adenocarcinoma. We followed-up 22 patients with ductal adenocarcinoma of the pancreas for a median of 19 months (range, 2 to 44 months). Angiogenesis was quantitated as vascular surface density (VSD) and the number of vessels per mm² stroma (NVES) after microvessels were immunostained, using factor VIII-related antigen. p53, H-ras, and Ki-67 proteins were also determined immunohistochemically. VSD and NVES showed significant correlations with increased proliferative activity, poor tumor differentiation, and tumor size of 3 cm or more (P = 0.001, P = 0.013, and P = 0.047, respectively). The overall 2-year survival rate of 33.3% in patients with high VSD and NVES values was significantly worse than that of 66.6% estimated in patients with low microvessel count (log rank, 3.97; P = 0.046). In multivariate analysis using the Cox model, VSD was found to be an independent prognostic factor of survival (P = 0.039). H-ras and p53 expressions were not correlated with angiogenesis parameters. We conclude that, in pancreatic ductal adenocarcinoma, angiogenesis is closely related to tumor growth and patient survival.     

Angiogenesis and antiangiogenic strategies in pancreatic cancer

Despite numerous advances in the treatment of solid tumors, the prognosis of patients diagnosed with pancreatic cancer remains dismal. Results of both surgical and non-surgical treatment for pancreatic cancer have been extremely disappointing beeause of the tumors propensity to metastasize, failure of chemotherapy to achieve adequate levels within the tumor, and resistance of pancreatic cancer to cytotoxic agents. Current chemotherapeutic agents and radiation treatments rely on the rapidly dividing nature of tumor cells and are limited by their cytotoxic effects on normal cells. The need to exploit the difference between normal and malignant cells has resulted in an enormous amount of research into the process of tumor neovascullarization. New agents are currently being developed that block tumor growth and metastasis through inhibition of angiogenesis. This article reviews the process of angiogenesis and antiangiogenic strategies with a special emphasis on pancreatic cancer.