Ursolic acid inhibits tumor angiogenesis and induces apoptosis through mitochondrial-dependent pathway in Ehrlich ascites carcinoma tumor

Ursolic acid (UA) is a pentacyclic triterpene naturally occurring in many plant foods. In the present study, we investigated anti-cancer activity of UA in vivo in Ehrlich ascites carcinoma (EAC) tumor. 15 × 10⁶ EAC cells were implanted intraperitoneally (i.p., ascitic tumor) and subcutaneous (s.c., solid tumor) in Swiss albino mice. Mice with established tumors received UA i.p. at 25, 50 and 100 mg/kg bw for 14 d in ascitic and 100 mg/kg bw in solid tumor for 30 d. On day 15, blood samples were collected for hematological assessment of hemoglobin (Hb%), RBCs, WBCs and PCV. Tumor volume, cell viability, angiogenic, anti-angiogenic, anti-inflammatory factors and antioxidant parameters were determined. Immunohistochemistry analysis for VEGF, iNOS, CD31, caspase-3 and Bax were also performed. UA significantly inhibited tumor growth, cell viability, in both ascites and solid tumor model in vivo (p < 0·001). The anti-angiogenic effects were accompanied with decreased VEGF, iNOS, TNF-α and increased IL-12 levels. UA at 100 mg/kg bw dose significantly increased SOD and CAT activity (p < 0.01). GSH and TBARS were increased as compared to control group (p < 0.001). Furthermore, UA increased total RBCs, WBCs as well as Hb% significantly (p < 0.05) compared to cyclophosphamide (CP). Histopathological examination of tumor cells in the treated group demonstrated signs of apoptosis with chromatin condensation and cell shrinkage. Decreased peritoneal angiogenesis showed the anti-angiogenic potential. UA downregulated VEGF & iNOS expression whereas bax and caspase-3 expressions were upregulated suggesting drug induced tumor cell apoptosis through activating the pro-apoptotic bcl-2 family and caspase-3 and downregulation of VEGF. The present study sheds light on the potent antitumor property of the UA and can be extended further to develop therapeutic protocols for treatment of cancer.     

Effect of weak combined static and extremely low‐frequency alternating magnetic fields on tumor growth in mice inoculated with the Ehrlich ascites carcinoma

It has been shown that the ultralow‐frequency extremely weak alternating component of combined magnetic fields (MFs) exhibits a marked antitumor activity. The parameters of this component have been found (frequency 1, 4.4, 16.5 Hz or the sum of these frequencies; intensity 300, 100, 150–300 nT, respectively) at which this MF in combination with a collinear static MF of 42 µT inhibits or suppresses the growth of Ehrlich ascites carcinoma (EAC) in mice. It was shown that the exposure of mice with EAC to combined MFs causes structural changes in some organs (liver, adrenal glands), which are probably due to the total degradation of the tumor tissue. In mice with transplanted EAC, the tumor tissue after exposure to weak MFs was practically absent, as distinct from control animals in which the invasion of the tumor into the adipose tissue surrounding the kidneys, mesenteric lymph nodes, and spermatic appendages was observed. In animals without tumors, no pathological deviations from the norm in the structure of organs and tissues occurred after exposure to weak MF, indicating that this factor per se is not toxic to the organism. Bioelectromagnetics 30:343–351, 2009. © 2009 Wiley‐Liss, Inc.     

肾癌患者血清VEGF的水平及其与临床病理分期的关系

目的:探讨肾癌患者血清血管内皮生长因子(VEGF)的水平及其与临床病理分期的关系。方法:选择2013年1月至2015年1月在我院行手术治疗的肾癌患者56例为观察组,选择同期在我院进行健康体检的正常成人50例作为对照,所有患者的诊断均经病理切片证实,对所有研究对象,采集其清晨空腹静脉血,用酶联免疫吸附法(ELISA)检测血清中的VEGF和VEGF受体-1(VEGFR-1)。结果:观察组血清中VEGF和VEGFR-1的浓度分别为(132.75±68.31)ng/mL和(33.76±15.39)ng/mL,均显著高于对照组,差异有统计学差异(均P0.05);不同分期患者血清中VEGF和VEGFR-1浓度差异有统计学意义(均P0.05),病理分期增加,VEGF与VEGFR-1的浓度增加,VEGF与VEGFR-1呈正相关(r=0.625,P0.05)。结论:血清VEGF水平可用于诊断RCC,且对于预判RCC的病例分期具有重要价值。     

Identification of Ca2+-pump-related phosphoprotein in plasma membrane vesicles of Ehrlich ascites carcinoma cells

Plasma membrane vesicles of Ehrlich ascites carcinoma cells have been isolated to a high degree of purity. In the presence of Mg²⁺, the plasma membrane preparation exhibits a Ca²⁺-dependent ATPase activity of 2 μmol P_i per h per mg protein. It is suggested that this $\text{(}\text{Ca}{}^{\mathrm{2+}}\text{+}\text{Mg}{}^{\mathrm{2+}}\text{)-}\text{ATPase}$ activity is related to the measured Ca²⁺ transport which was characterized by $\text{K}{}_{\mathrm{<mtext>m</mtext>}}$ values for ATP and Ca²⁺ of $\text{44 ± 9 μ}\text{M}$ and $\text{0.25 ± 0.10 μ}\text{M}$, respectively. Phosphorylation of plasma membranes with [γ-³²P]ATP and analysis of the radioactive species by polyacrylamide gel electrophoresis revealed a Ca²⁺-dependent hydroxylamine-sensitive phosphoprotein with a molecular mass of 135 kDa. Molecular mass and other data differentiate this phosphoprotein from the catalytic subunit of $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ and from the catalytic subunit of $\text{(}\text{Ca}{}^{\mathrm{2+}}\text{+}\text{Mg}{}^{\mathrm{2+}}\text{)-}\text{ATPase}$ of endoplasmic reticulum. It is suggested that the 135 kDa phosphoprotein represents the phosphorylated catalytic subunit of the $\text{(}\text{Ca}{}^{\mathrm{2+}}\text{+}\text{Mg}{}^{\mathrm{2+}}\text{)-}\text{ATPase}$ of the plasma membrane of Ehrlich ascites carcinoma cells. This finding is discussed in relation to previous attempts to identify a Ca²⁺-pump in plasma membranes isolated from nucleated cells.     

VEGF-C和VEGFR-3在胃癌组织中的表达及临床意义

目的：探讨胃癌患者血管内皮生长因子C（vascular endothelial growth factor-C,VEGF-C及血管内皮生长因子受体-3（vascular endothelial growth factor receptor-3,VEGFR-3）在胃癌组织中的表达,从而确定胃癌预后的分子标志物。方法：搜集整理临床资料,采用Real-time PCR及ELISA法检测43例胃癌组织VEGF-C和VEGFR-3的表达。结果：43例胃癌组织中均有不同程度的VEGF-C和VEGFR-3的表达,Real-time PCR结果显示胃癌组织淋巴结转移组和非转移组VEGF-C和VEGFR-3的表达分别为0.07±0.01和0.12±0.01,0.03±0.01和0.06±0.02,与正常对照组相比,差异有显著性（p〈0.05）。ELISA检测显示,与正常胃组织中VEGF-C和VEGFR-3的蛋白表达相比,胃癌无淋巴结转移组及胃癌并发淋巴结转移组中VEGF-C和VEGFR-3均明显增加。结论：VEGF-C和VEGFR-3的表达与胃癌淋巴结转移密切相关,提示胃癌标本VEGF-C和VEGFR-3的检测可作为胃癌预后的分子标志物。     

VEGF-C 和VEGFR-3 在胃癌组织中的表达及临床意义

目的:探讨胃癌患者血管内皮生长因子C(vascular endothelial growth factor-C,VEGF-C及血管内皮生长因子受体-3(vascular endothelial growth factor receptor-3,VEGFR-3)在胃癌组织中的表达,从而确定胃癌预后的分子标志物。方法:搜集整理临床资料,采用Real-time PCR及ELISA法检测43例胃癌组织VEGF-C和VEGFR-3的表达。结果:43例胃癌组织中均有不同程度的VEGF-C和VEGFR-3的表达,Real-time PCR结果显示胃癌组织淋巴结转移组和非转移组VEGF-C和VEGFR-3的表达分别为0.07±0.01和0.12±0.01,0.03±0.01和0.06±0.02,与正常对照组相比,差异有显著性(p<0.05)。ELISA检测显示,与正常胃组织中VEGF-C和VEGFR-3的蛋白表达相比,胃癌无淋巴结转移组及胃癌并发淋巴结转移组中VEGF-C和VEGFR-3均明显增加。结论:VEGF-C和VEGFR-3的表达与胃癌淋巴结转移密切相关,提示胃癌标本VEGF-C和VEGFR-3的检测可作为胃癌预后的分子标志物。     

A flavone from the ethyl acetate extract of Leea rubra leaves with DNA damage protection and antineoplastic activity

Among the major constituents of Leea rubra (Family Vitaceae) leaves, phenolic and flavonoind compounds are most important for therapeutic purposes and the plant parts have been used in traditional medicine to treat several diseases for long. Thus, in order to scientifically confirm the traditional uses of the L. rubra leaves, the present study was designed to investigate the efficacy of the isolated flavones against AAPH induced oxidative damage to pUC19 DNA by gel electrophoresis and antineoplastic activity was evaluated on Ehrlich ascites carcinoma (EAC) bearing Swiss albino mice by evaluating percentage inhibition of cell growth, morphological changes of EAC cells and hematological parameters of the mice. The isolation was carried out by column chromatography and structure was revealed by ¹H-NMR and ¹³C NMR. The result shows that, the isolated compound was identified as myricetin 4'-methoxy-3-O-α-l-rhamnopyranoside based on previously reported data. The isolated flavone effectively inhibited AAPH-induced oxidative damage to DNA; because it could inhibit the formation of circular and linear forms of the DNA. In anti-proliferative assay, 76% growth inhibition of EAC cells was observed as compare to the control mice (p<0.05) at a dose 100 mg/kg body weight. Thus the isolated flavone showed great importance as a possible therapeutic agent in preventing oxidative damage to DNA and the chronic diseases caused by such DNA damage, and can also become important in cancer chemotherapy.     

Translation in micrococcal nuclease-treated cell-free extracts from Ehrlich ascites tumor cells: Stimulation by initiation factor eIF-2B

Translation of exogenous mRNAs in micrococcal nuclease-treated extracts from Ehrlich ascites tumor cells is greatly stimulated by the addition of crude initiation factors or initiation factors eIF-2B and eIF-2 containing eIF-2B. The requirement for exogenous eIF-2B in micrococcal nuclease-treated extracts does not result from either loss or enhanced phosphorylation of eIF-2 during incubation.     

Mechanisms mediating metabolic abnormalities in the livers of Ehrlich ascites tumor-bearing mice

Previously we reported that intermittent intraperitoneal administration of ornithine decarboxylase-inducing factor (ODC factor), interleukin 1alpha (IL-1alpha), and tumor-necrosis factor-alpha (TNF-alpha) to normal mice induced biological changes in the hosts which included changes in the pattern of expression of pyruvate kinase (PK) isozymes in the liver and hypertrophy of the spleen. In the study reported here, we investigated the chronic and combined effects of these factors on hepatic enzymes using alzet microosmotic pumps implanted in the subcutis of the backs or abdominal cavities of mice. Continuous administration of ODC factor and recombinant human IL-1alpha (rhIL-1alpha) reduced the activity of L-type PK, which is a glycolysis-related enzyme in the liver, and induced the activity of M2-type PK, a known marker of liver dedifferentiation. Serine dehydratase (SDH) and tyrosine aminotransferase (TAT), enzymes associated with amino acid metabolism, were not significantly influenced at the examined concentration. The simultaneous continuous infusion of ODC factor and rhIL-1alpha or rhTNF-alpha caused alterations in the patterns of expression of PK isozyme activity profiles and reduced overall PK activity. SDH and TAT activities were also significantly induced. Moreover, mice treated with these combined factors displayed many other metabolic changes normally associated with cancer cachexia. These findings suggest that the tumor-derived ODC factor and cytokines such as IL-1alpha and TNF-alpha might function synergistically in the metabolic perturbations observed in Ehrlich ascites tumor bearers.     

Punarnavine,an alkaloid from Boerhaavia diffusa exhibits anti-angiogenic activity via downregulation of VEGF in vitro and in vivo

Punarnavine, a quinolizidine alkaloid isolated from Boerhaavia diffusa is known to possess analgesic, anti-inflammatory, hepato-protective, immunomodulatory and anti-proliferative properties. However, its roles in tumor angiogenesis and the involved molecular mechanism are still unknown. Therefore, we examined its anti-angiogenic effects and mechanisms in vitro and in vivo. We examined the effect of punarnavine on VEGF-A expression by RT-PCR, Western blotting and ELISA. In vivo antiangiogenic activity was determined using sponge implant angiogenesis assay and antitumor activity was evaluated against Ehrlich ascites carcinoma tumor. Punarnavine significantly inhibited endothelial cell migration and invasion and capillary structure formation of HUVECs. Punarnavine significantly at 50 μM inhibited MMP-2 and MMP-9 expression in HUVECs in vitro. Punarnavine inhibited neovascularization in sponge implant assay. Punarnavine (15 mg/kg bw/d) treatment showed dose-dependent decrease in the ascitic fluid volume by 60.94% and tumor volume by 86.40% in Ehrlich ascites model. Reduction in peritoneal angiogenesis with punarnavine treatment suggests the anti-angiogenic activity of punarnavine. The present study sheds light on the potent anti-angiogenic of the punarnavine and can be extended further to develop therapeutic protocols for treatment of cancer.     

甲状腺乳头状癌中Survivin,VEGF,EGFR的表达及临床意义分析

目的:探讨甲状腺乳头状癌(PTC)中存活素(Survivin)、血管内皮生长因子(VEGF)、表皮生长因子受体(EGFR)的表达及临床意义。方法:选择2013年4月~2016年4月在我院收治的100例PTC患者的手术标本以及同期100例癌旁正常甲状腺组织为研究对象,采用免疫组化方法检测盒比较Survivin、VEGF、EGFR的表达在PTC及癌旁正常甲状腺组织中的表达及二者的相关性,并分析其Survivin、VEGF、EGFR的表达与PTC患者临床病理特征的相关性。结果:PTC组织中Survivin、VEGF、EGFR阳性表达率均显著高于癌旁正常甲状腺组织,差异具有统计学意义(P0.05);Survivin、VEGF在淋巴结转移患者中的阳性表达率显著高于无淋巴结转移者(P0.05);EGFR在女性PTC患者中的阳性表达率显著高于男性(P0.05)。PTC组织中VEGF和Survivin、EGFR的表达无显著相关性(P0.05),但Survivin和EGFR的表达显著相关(r=0.235,P0.05)。结论:Survivin、VEGF、EGFR在PTC组织中表达上调,Survivin、VEGF与PTC的淋巴结转移有关,Survivin和EGFR在PTC的发生、发展过程中可能存在着协同作用。     

Interleukin-6 increases vascular endothelial growth factor and angiogenesis in gastric carcinoma

Interleukin-6 (IL-6) is a proinflammatory cytokine associated with the disease status of gastric carcinoma (GC). Vascular endothelial growth factor (VEGF) is a potent tumor angiogenic factor in GC. In this study, we attempted to clarify whether IL-6 can regulate VEGF and angiogenesis in GC. GC samples from 54 surgical specimens were subjected to immunohistochemical examination of IL-6, VEGF, and tumor microvessels, and results showed that IL-6 was positively correlated with VEGF expression and tumor vasculature. We determined VEGF expression in four GC cell lines by ELISA, revealing that GC cells can produce significant amount of VEGF with increasing dose and duration of IL-6 stimulation. Next, a luciferase reporter gene assay was employed to determine the signaling pathway driving the VEGF promoter by IL-6, which showed that the JAK/STAT pathway is involved in the stimulation of VEGF gene expression. The effects of IL-6 on angiogenesis in vitro and in vivo were evaluated by HUVEC studies and the Matrigel plug assay, respectively. Results showed that IL-6 effectively promoted HUVEC proliferation and tube formation in vitro and Matrigel plug vascularization in vivo, primarily by inducing VEGF in GC. This study provides evidence that the multifunctional cytokine, IL-6, may induce VEGF expression which increases angiogenesis in gastric carcinogenesis.     

Covalent binding of 1-nitro-9-(3-dimethyl-n-propylamino) acridine, a new antitumor drug, to DNA of Ehrlich ascites tumor cells in vivo.

After exposing a line of rat liver epithelial cells to a single dose of the carcinogen N-acetoxy-2-acetylaminofluorene (N-acetoxy-AAF), a dose-dependent decrease in [³H]uridine incorporation into total cellular RNA was found. Approx. 50% inhibition occurred with 0.5 μg/ml of the compound. The kinetics of the response, the effects of actinomycin D, and the fractionation of the newly synthesized RNA by polyacrylamide gel electrophoresis indicated preferential inhibition of the synthesis of 45S ribosomal RNA precursor and a relative sparing of the synthesis of heterogeneous nuclear RNA.     

血管形成在双歧杆菌预防大肠癌生长中的作用

目的 从血管形成角度探索青春型双歧杆菌预防大肠癌生长的途径。方法 建立大肠癌裸鼠移植瘤模型,以免疫组化法检测大肠癌组织血管内皮生长因子（VEGF）的蛋白表达水平及其微血管密度（MVD）。结果 双歧杆菌预防组大肠癌VEGF的阳性细胞密度及MVD的数量均明显低于肿瘤对照组（P〈0．01）。结论 青春型双歧杆菌能下调大肠癌VEGF的表达,进而抑制其血管形成,这可能是它预防大肠癌生长的途径之一。     

血管内皮生长因子在鼻咽癌病情监测中的意义

目的:探讨鼻咽癌患者治疗前后血清血管内皮生长因子(VEGF)水平的变化及其临床意义。方法:ELISA法对75例鼻咽癌患者在治疗前、治疗结束后1个月和治疗后出现局部复发或远处转移者,同步检测40例慢性鼻咽炎患者及30名正常人血清VEGF水平。结果:鼻咽癌组Ⅲ～Ⅳ期患者治疗前血清VEGF水平均显著高于Ⅰ～Ⅱ期患者(P<0.01);与治疗前比较,Ⅰ～Ⅱ期、Ⅲ～Ⅳ期患者治疗后血清VEGF水平均显著下降(P<0.01);治疗后,Ⅰ～Ⅱ期与Ⅲ～Ⅳ期患者血清VEGF水平比较,差异无统计学意义;与慢性鼻咽组及正常对照组比较,鼻咽癌复发组患者治疗前、后的血清VEGF水平明显升高(P<0.01);与治疗前组比较,鼻咽癌患者治疗后的血清VEGF水平明显下降(P<0.01),而复发组治疗后明显上升(P<0.01)。结论:血清VEGF水平可作为鼻咽癌患者治疗前后病情监测一个新的检测指标。     

Effect of 2,3-dinitrilo-1,4-dithia-9,10-antraquinone on Ehrlich ascites carcinoma and yeast cells

The inhibition of glycolysis by 2,3-dinitrilo-1,4-dithia-9,10-antraquinone (DDA) in Ehrlich ascites carcinoma (EAC) cells as well as in the investigated respiratory and fermentative strains of yeasts was found to be the result of inactivation of thiol enzymes of this pathway. Increasing concentration of DDA caused, in EAC cells, marked inhibition of hexokinase (HK), phosphofructokinase (PFK) and practically total inhibition of glyceraldehyde-3-phosphate dehydrogenase (GAPDH). These three enzymes, as well as alcohol dehydrogenase (ADH) were also inactivated by DDA in yeasts. DDA inhibited the biosynthetic processes as measured by following the rate of [14C]adenine and [14C)]valine incorporation into TCA-precipitable fractions proportionally to the degree of glucose consumption by EAC or the yeast cells.     

VEGF和Survivin在肾癌中的表达及其相关性研究

目的:探讨肾癌组织中血管内皮生长因子VEGF与凋亡抑制蛋白Survivin表达的相关性及其之间的关系,研究Survivin和VEGF在肾癌发生发展中的作用机制。方法:应用免疫组织化学方法检测70例肾癌组织和70例癌旁正常肾脏组织中VEGF和Survivin的表达,并将检测结果与临床病理特征进行综合分析。结果:VEGF和Survivin在肾癌中表达均高于癌旁正常肾脏组织;Survivin和VEGF在肾癌中的阳性表达率分别为75.71%(53/70)和72.86%(51/70),在癌旁肾脏组织中的表达率分别为0%(0/70)、17.14%(12/70),差异均有显著性意义(P0.05);VEGF和Survivin的表达与患者的性别、年龄、肿瘤大小、病理分级均无相关性;VEGF和Survivin表达呈正相关性。结论:VEGF和Survivin在肾癌组织中表达率较高,为肾癌的分子靶向治疗提供了新的靶点。Survivin和VEGF在RCC中的表达关系密切,测定RCC中Survivin、VEGF蛋白的表达,有助于临床判断病人预后。     

鼻息肉与鼻咽癌标本VEGF水平及微血管密度对比及其临床意义

目的:对比研究鼻咽癌和鼻息肉标本中VEGF表达强度及MVD差异,同时分析VEGF、MVD和鼻咽癌临床特征的相关性。方法:纳入我科就诊的鼻咽癌患者57例,鼻息肉患者50例。采用免疫组化SABC法检测癌组织、癌旁组织、及息肉组织中中VEGF蛋白的表达,及MVD强度。分析VEGF、MVD和鼻咽癌患者性别、临床分期、颈部淋巴结转移、远处转移、血清EBV-Ig A阳性、WHO病理分型相关性。统计分析随访结果,对可能影响鼻咽癌预后的因素进行Cox回归模型分析。结果:鼻咽癌组织、鼻咽癌旁组织、鼻息肉组织中VEGF表达、MVD强度具有明显差异(p0.05)。不同鼻咽癌临床分期、是否发生远处转移、不同WHO病理分型和VEGF表达、MVD强度具有明显差异(p0.05)。Cox回归方程显示,远处转移、病理分型、VEGF表达强度是影响鼻咽癌生存的独立危险因素(p0.05)。结论:鼻咽癌高表达VEGF,促进新生血管,形成高密度微小血管,和鼻咽癌远处转移密切相关,降低其生存率。     

Novel VEGFR-2 Kinase Inhibitors as Anticancer Agents: A Review Focusing on SAR and Molecular Docking Studies (2016–2021)

Cancer growth, annexation, and metastatic spread are all aided by the formation of new blood vessels (angiogenesis). The commencement of the VEGF pathway leads to signal transduction that enhances endothelial cell survival, relocation, and divergence from pre-existing vasculature. The ability of solid malignancies to bloom and spread depends critically on their ability to establish their independent blood circulation (tumor angiogenesis). VEGFR is a major receptor tyrosine kinase that regulates angiogenesis, cell growth, and metastasis, diminishing apoptosis, cytoskeletal function, and other biological processes VEGFR has proven to be a remarkable focus for a variety of anticancer medicines in clinical studies. This Review explores the development of anti-VEGF-based antiangiogenic therapies having different scaffolds. This review had focused on SAR and docking studies of previously reported molecules.     

KDP胞外502～764位氨基酸基因合成、表达及功能鉴定

应用基因搭桥法及 Taq酶聚合反应合成了编码人血管内皮生长因子受体 - ( h VEGFR- ,KDR)第 50 2～ 764位 2 62个氨基酸的基因片段 .DNA序列分析表明 ,合成的 786bp的基因片段与文献报道的 KDR相应 c DNA序列完全一致 .将该基因与原核融合蛋白表达载体 p GEX- 3X重组 ,在大肠杆菌 JM1 0 9中表达了 GST- KDR2 62融合蛋白 ,表达量约占菌体总蛋白的 35% .表达产物依次经包涵体分离、变性、复性、亲合层析纯化和 Xa因子酶解 ,获得了 KDR2 62目的蛋白纯品 .GST-KDR2 62融合蛋白和纯化产物经 Western blot分析 ,两者均可被 VEGF1 65特异性识别 ,前者分子量约 56k D,后者分子量约 30 k D;这两种蛋白用 VEGF1 65及其抗体进行的 ELISA分析结果均显示阳性 ,并有剂量依赖关系 ,而用 Xa因子酶解 GST- KDR2 62融合蛋白获得的 GST和空载体诱导产物对照均为阴性 .以上结果表明表达的 KDR2 62蛋白可特异性地与 VEGF结合 .