“Real life” polycyclic aromatic hydrocarbon (PAH) mixtures modulate hCG,hPL and hPLGF levels and disrupt the physiological ratio of MMP-2 to MMP-9 and VEGF expression in human placenta cell lines

Using JEG-3 and BeWo cells, we examined the effect of “real life” mixtures of polycyclic aromatic hydrocarbons (PAHs), at doses reported in maternal blood (Mix I) and in placental tissue (Mix II), on human chorionic gonadotropin (hCG), placental lactogen (hPL) and placental growth factor (hPLGF) secretion, protein expression and immunolocalization. Additionally, the action of PAH mixtures on basal and hormone-stimulated matrix metalloproteinase-2 (MMP-2), MMP-9 and vascular endothelial growth factor (VEGF) protein expression was evaluated. Under basal conditions, the PAH mixtures increased hCG and decreased hPLGF levels in both cell lines, while hPL expression was stimulated in JEG-3 and inhibited in BeWo. There was no effect on the MMP-2/MMP-9 ratio or VEGF expression. In hormone-stimulated cells, PAH mixtures changed the MMP-2/MMP-9 ratio in JEG-3 cells in favor of MMP-9, while in BeWo MMP-2 was favored. The effect on VEGF expression was cell specific and dependent on the mixture. In hCG-treated cells, only Mix II inhibited VEGF expression in JEG-3 cells. Neither PAH mixtures affected this protein in BeWo cells. In hPL-treated cells, Mix I had a stimulatory effect in JEG-3 cells, while Mix II exerted an inhibitory effect in BeWo cells. In hPLGF-treated cells, Mix II decreased in JEG-3 cells, but in BeWo cells, both mixtures increased VEGF expression. Considering that the evaluated protein hormones play crucial roles in angiogenesis and neovascularization in the placenta, “real life” PAH mixtures by disrupting protein hormones levels, the MMP-2/MMP-9 ratio and VEGF expression can lead to insufficiency and many pregnancy-related disorders.     

A DNA vaccine expressing an optimized secreted FAPα induces enhanced anti-tumor activity by altering the tumor microenvironment in a murine model of breast cancer

Cancer-associated fibroblasts (CAFs), major components of the tumor microenvironment (TME), promote tumor growth and metastasis and inhibit the anti-tumor immune response. We previously constructed a DNA vaccine expressing human FAPα, which is highly expressed by CAFs, to target these cells in the TME, and observed limited anti-tumor effects in the 4T1 breast cancer model. When the treatment time was delayed until tumor nodes formed, the anti-tumor effect of the vaccine completely disappeared. In this study, to improve the safety and efficacy, we constructed a new FAPα-targeted vaccine containing only the extracellular domain of human FAPα with a tissue plasminogen activator signal sequence for enhanced antigen secretion and immunogenicity. The number of CAFs was more effectively reduced by CD8⁺ T cells induced by the new vaccine. This resulted in decreases in CCL2 and CXCL12 expression, leading to a significant decrease in the ratio of myeloid-derived suppressor cells in the TME. Moreover, when mice were treated after the establishment of tumors, the vaccine could still delay tumor growth. To facilitate the future application of the vaccine in clinical trials, we further optimized the gene codons and reduced the homology between the vaccine and the original sequence, which may be convenient for evaluating the vaccine distribution in the human body. These results indicated that the new FAPα-targeted vaccine expressing an optimized secreted human FAPα induced enhanced anti-tumor activity by reducing the number of FAPα⁺ CAFs and enhancing the recruitment of effector T cells in the 4T1 tumor model mice.     

NRP-1单克隆抗体对乳腺癌裸鼠移植瘤生长的影响

 目的 探讨NRP-1单克隆抗体（NRP-1 MAb）的特异性，以及不同剂量的NRP-1 MAb治疗乳腺癌裸鼠移植瘤的疗效。方法 Western blot和共聚焦免疫荧光法检测NRP-1 MAb是否识别MCF7细胞上NRP-1蛋白。将MCF7细胞接种于BALB/c裸鼠皮下建立乳腺癌细胞移植瘤模型，并进行瘤组织传代。传代的肿瘤体积生长至300~500 mm3时，随机分为对照组、NRP-1 MAb低剂量组、中剂量组和高剂量组，每组6只，给药7次。观察荷瘤裸鼠一般状况，测量瘤体大小及裸鼠体重。实验结束时剥离瘤体称重，提取组织蛋白，Western blot检测组织中VEGF蛋白和NRP-1蛋白的表达量。结果 NRP-1MAb成功识别MCF7细胞上的NRP-1蛋白；NRP-1 MAb能够有效抑制MCF7细胞裸鼠移植瘤的生长，低剂量组（1 mg/kg）抑瘤率为47.01%，中剂量组（5 mg/kg）抑瘤率为65.70%，高剂量组（10 mg/kg）抑瘤率为69.19%。。结论 NRP-1 MAb能够识别并有效结合MCF7细胞膜上的NRP-1蛋白，且可抑制MCF7细胞移植瘤的生长，NRP-1 MAb抑制移植瘤的增长可能与下调NRP-1和VEGF表达有关。     

膀胱移行细胞癌组织中VEGF和PCNA表达的关系及其临床意义

目的:研究血管内皮生长因子(VEGF)和增殖细胞核抗原(PCNA)在同一膀胱移行细胞癌(BTCC)组织中的表达及其与BTCC临床、病理的关系,探讨二者与肿瘤血管形成和肿瘤细胞增殖的关系。方法:采用免疫组织化学LDP法对77例BTCC组织中VEGF和PCNA的表达情况进行检测。结果:BTCC组织中VEGF与PCNA阳性表达率分别为66.23%和51.94%,VEGF表达和PCNA表达均与BTCC的组织学分级显著相关(P<0.05);浸润性肿瘤明显高于表浅性肿瘤(P<0.05);术后复发组明显高于未复发组(P<0.01);在VEGF阳性表达的病例中,PCNA表达随VEGF表达强度的升高而升高(P<0.05),且肿瘤复发率也升高,二者呈明显正相关性(P<0.05)。VEGF和PCNA阳性或同时表达阳性的患者复发迅速,PCNA阴性而VEGF阳性表达者较阴性表达者预后差。结论:VEGF和PCNA与BTCC组织学分级、恶性程度和预后密切相关。BTCC是典型的血管依赖性和增殖性肿瘤。VEGF和PCNA均可能通过不同的途径参与了肿瘤血管形成和肿瘤细胞增殖的调控。     

血管内皮生长因子反义RNA对C6鼠胶质瘤细胞体外作用的研究

目的探讨VEGF反义RNA对C6鼠胶质瘤细胞体外作用。方法用Lipofectamine介导的方法将VEGF反义eDNA转染入C6鼠胶质瘤细胞系,观察其对细胞增殖和凋亡的影响。结果在体外VEGF反义RNA可有效抑制C6鼠胶质瘤细胞内源性VEGFmRNA和VEGF蛋白的产生,但对细胞增殖和凋亡无明显影响。结论采用VEGF反义RNA可能成为抗血管形成肿瘤治疗的新策略之一。     

血管内皮生长因子及炎性因子在油酸性犬急性呼吸窘迫综合征中的变化

目的探讨油酸性急性呼吸窘迫综合征（ARDS）beagle犬血浆及肺泡灌洗液血管内皮生长因子（VEGF）、可溶性细胞间黏附分子-1（ICAM-1）、白细胞介素-8（IL-8）、肿瘤坏死因子-α（TNF—α）水平的改变。方法12只英国纯种beagle犬，静脉注射油酸0．15mL／kg，在注射油酸前、后1h，出现ARDS的典型表现时，抽血测VEGF、sICAM-1、IL-8、TNF—α，并对此时相作肺泡灌洗液VEGF、sICAM—1、IL-8、TNF-α的测定。结果beagle犬静脉注射油酸后1h血浆TNF—α升高（P〈0．05），血浆反肺泡灌洗液IL8、sICAM-1和VEGF在1h较油酸前没有明显变化（P〉0．05），beagle犬油酸型ARDS模型建立后血浆及肺泡灌洗液VECF、sICAM-1、IL-8、TNF—α均显著高于建模前（P〈0．05）。结论VEGF、sICAM-1、IL-8、TNF～α在beagle犬油酸型ARDS发生发展过程中可能均起重要作用，其水平的高低可能与ARDS严重程度及预后有关。     

大鼠缺血后肢骨骼肌血管内皮生长因子及其受体的表达

目的 研究大鼠缺血后肢侧枝代偿和血管内皮生长因子（VEGF）及其受体表达的动态变化。为外源性VEGF治疗下肢缺血性疾病提供理论依据。方法 切除SD大白鼠右后肢全长股动脉，随机分为9个时间组：造模后1、3d、1、2、3、4、6、8及12周，各组5只动物。分别于造模前后和观察期末检测双后肢大、小腿肌肉Fit-1、Flk-1蛋白及mRNA表达，各组观察期末实验动物后肢动脉DSA检查。结果 （1）缺血后3d，5只大鼠右后肢出现溃疡（11．11％）；2周后，4只大鼠后肢溃疡愈合，而1只趾端坏疽（2．22％）。（2）缺血后2周，患肢侧枝形成达到高峰，12周时仍可见侧支血管显影。（3）缺血早期（3周内），VEGF及其受体的表达均较健侧显著增强（P〈0．05）；缺血中期（3～8周）。VEGF和Flt-1表达迅速下降，Flk-1仍表达；缺血后期（8周后），VEGF及其受体的表达均低至极低水平，与对侧差异无统计学意义（P〉0．05）。结论（1）肢体缺血后自身的血管新生不能完全满足缺血组织的需要。（2）缺血早期外源性的VEGF补充是不必要的；缺血中期补充VEGF是适宜的；缺血后期在应用VEGF治疗的同时，也需要干预受体的表达。     

VEGF在膀胱移行细胞癌中的表达及意义

目的：探讨膀胱移行细胞癌中血管内皮生长因子（VEGF）的表达及其与临床病理指标的关系。方法：应用免疫组化方法对40例膀胱移行细胞癌及15例正常膀胱组织中VEGF进行检测。结果：正常膀胱移行上皮均为阴性表达；膀胱移行细胞癌中VEGF阳性表达24例，表达率为60％（24／40），明显高于正常膀胱组织（P＜0.01），其中VEGF表达阳性率随膀胱癌病理分级、临床分期上升而升高，表达强度也随之增强（P＜0.01）。结论：VEGF表达对膀胱移行细胞癌生物学行为有重要影响，VEGF有可能可作为判断膀胱癌生物学行为、转移潜能及预后指标。     

A comparative study of angiogenic and cytokine responses after laparoscopic cholecystectomy performed with standard- and low-pressure pneumoperitoneum

Background  Surgical procedures enhance production of pro- and anti-inflammatory cytokines and angiogenic factors that play a pivotal role in the immunological response to surgical trauma and take part in the pathogenesis of tumor growth and adhesions formation. The purpose of the study was to access the influence of low-pressure CO₂ pneumoperitoneum on the inflammatory and angiogenic responses during the postoperative period after laparoscopy. Methods  The study group consisted of 40 patients, operated on due to cholelithiasis using standard-pressure (n = 20) and low-pressure (n = 20) CO₂ pneumoperitoneum. Serum concentration of interleukin (IL)-6, IL-8, IL-10, vascular endothelial growth factor (VEGF)-A, and endostatin were measured before and at 6, 24, and 48 h after surgery with commercially available enzyme-linked immunosorbent assay (ELISA). Results  Concentrations of IL-6 increased significantly after the operations in both groups. No differences were observed between the groups in regards to IL-6, IL-8, and IL-10 levels. Concentrations of VEGF-A measured at 6 and 48 h were significantly lower in patients who underwent laparoscopies performed with low-pressure pneumoperitoneum. No significant variations were observed in endostatin serum concentration. Concentrations of the studied parameters were not influenced by duration of surgery or by age, gender, or body mass index (BMI) of the patients. Conclusions  The results obtained in our study do not show any significant differences between studied operative procedures with regards to systemic inflammatory response. Changes in the concentrations of VEGF-A and endostatin observed in the studied population may suggest this technique is more favorable with regards to angiogenesis process intensity, along with all its consequences and implications.