Proteomic tracking of serum protein isoforms as screening biomarkers of ovarian cancer

Epithelial ovarian cancer is the fourth leading cause of cancer death among women. Due to the asymptomatic nature and poor survival characteristic of the disease, screening for specific biomarkers for ovarian cancer is a major health priority. Differentially expressed proteins in the serum of ovarian cancer patients have the potential to be used as cancer-specific biomarkers. In this study, proteomic methods were used to screen 24 serum samples from women with high-grade ovarian cancer and compared to a control group of 11 healthy women. Affigel-Blue treated serum samples were processed either by linear (pH 4-7) or narrow range (pH 5.5-6.7) IEF strips for the first dimension. Proteins separated in first dimension were resolved by 8-16% gradient SDS-PAGE. Protein spots were visualized by SYPRO Ruby staining, imaged by FX-imager and compared and analyzed by PDQuest software. Twenty-two protein spots were consistently differentially expressed between normal and ovarian cancer patients by resolving proteins in a linear pH strip of 4-7 for the first dimension. Six of the protein spots, significantly up-regulated in grade 3 ovarian cancer patients (p < 0.05), were identified by MALDI-TOF MS and Western blotting as the isoforms of haptoglobin precursor. When serum proteins were resolved on narrow pH range strips (5.5-6.7), 23 spots were consistently differentially expressed between normal and grade 3 ovarian cancer patients. Of these, 4 protein spots significantly down regulated in grade 3 ovarian cancer patients (p < 0.05) were identified by MALDI-TOF MS and Western blotting, as isoforms of transferrin precursor. Increased expression of serum haptoglobin and transferrin was also identified in peritoneal tumor fluid obtained from women diagnosed with grade 2/3 ovarian cancer (n = 7). Changes in the expression of haptoglobin and transferrin in the serum of women with different pathological grades of ovarian cancer was examined by one-dimensional Western blotting method. Serum samples collected from women suffering from benign, borderline, grade 1, grade 2 and grade 3 cancer (n = 4 for haptoglobin and n = 5 for transferrin in each group) were analyzed and compared to the serum of normal healthy women. The mean serum haptoglobin expression in grade 3 ovarian cancer patients was fourfold higher than in the control subjects (p < 0.05). On the other hand, transferrin expression in grade 3 ovarian cancer patients was decreased by twofold than in normal healthy women (p < 0.05). Haptoglobin expression in the serum of cancer patients (n = 7) decreased following chemotherapy (six cycles of taxol/carboplatin). Concomitant with the decrease of haptoglobin, transferrin expression remained constant in four patients, but increased in three out of seven patients included in the study. Changes in serum expression of haptoglobin correlated with the change of CA 125 levels before and after chemotherapy. In conclusion, proteomic profiling of differentially expressed proteins in the sera of normal women compared to women with ovarian cancer can greatly facilitate the discovery of a panel of biomarkers that may aid in the detection of ovarian cancer with greater specificity.     

卵巢癌患者血清HE4、CA125、CA72-4及炎性因子水平检测的临床价值评估

目的:探讨血清人附睾蛋白4(HE4)、糖类抗原125(CA125)、糖类抗原72-4(CA72-4)及炎性因子IL-6、IL-8、IL-17水平检测对卵巢癌患者的临床价值。方法:选取2015年2月至2017年2月我院收治的卵巢肿瘤患者81例,包括卵巢恶性肿瘤组39例及卵巢良性肿瘤组42例,另选42例健康人作正常对照组,比较各组及临床不同分期卵巢癌患者血清HE4、CA125、CA72-4、IL-6、IL-8、IL-17水平的差异。结果:卵巢恶性肿瘤组血清HE4、CA125、CA72-4、IL-6、IL-8及IL-17水平均显著高于对照组(P0.05),而卵巢良性肿瘤组与对照组以上指标比较差异并无统计学意义(P0.05);随卵巢癌分期增加,患者血清HE4、CA125、CA72-4、IL-6、IL-8及IL-17水平呈上升趋势,以血清HE4、CA125及IL-17增加较为显著(P0.05),而Ⅰ期、Ⅱ期及Ⅲ期卵巢癌患者血清CA72-4、IL-6及IL-8在增加并不明显(P0.05)。结论:卵巢癌患者血清HE4、CA125、CA72-4、IL-6、IL-8及IL-17水平较高,且随临床分期增加呈上升趋势,以上指标可能有助于卵巢癌的诊断、治疗及病情评估。     

Prognostic value of IL-27 polymorphisms and the susceptibility to epithelial ovarian cancer in a Chinese population

This study investigated the association between IL-27 gene polymorphisms and susceptibility to epithelial ovarian cancer in a Chinese population and discusses the risk factors associated with survival time. We collected data on 229 patients diagnosed with epithelial ovarian cancer, from 15 to 77 years of age with a long clinical follow-up period. Polymerase chain reaction-restriction fragment length polymorphism was performed to determine the genotype of IL-27 gene polymorphisms. Ovarian cancer-specific survival (OCSS) according to genotype of IL-27 gene polymorphisms was explored by Kaplan–Meier analysis and Cox proportional hazards modeling. Significant differences for genotype frequencies of both SNP sites were found between cases and controls. Both allele G frequencies were significantly greater among the cases (rs153109: 0.404 vs. 0.303, P = 0.001, odds ratio [OR]?=?1.333, 95% confidence interval [CI]?=?1.133–1.567; rs17855750: 0.146 vs. 0.083, P = 0.001, OR?=?1.766, 95% CI?=?1.258–2.481). Haplotype analysis showed haplotypes AG, GT and GG were associated with increased ovarian cancer susceptibility while AT was a protective haplotype. Advanced FIGO stage (stages III?+?IV) and non-optimal cytoreductive surgery (residual tumor ≥1 cm) were poor prognostic factors in the univariate analysis (P = 0.003, P = 0.049). However, FIGO stage was found to be the only independent significant prognostic factor by Cox proportional hazards analysis (P = 0.042). IL-27p28 mRNA expression was significantly decreased in ovarian cancer patients (P?<?0.0001), while no significant relationship was found between IL-27p28 mRNA expression and polymorphism of rs153109 and rs17855750 (P?=?0.193 and P?=?0.146, respectively). Our study suggests that IL-27 gene polymorphisms may be involved in the susceptibility to epithelial ovarian cancer, but not in survival in a clinic-based Chinese population. Haplotype analysis of these two SNPs seems to be an important mark to predict the disease susceptibility. Advanced FIGO stage, as the only significant, independent risk factor, predicts poor clinical outcomes for patients diagnosed with epithelial ovarian cancer. The decreased expression of IL-27p28 mRNA in ovarian cancer might indicate the antitumor activities of this novel cytokine.     

Involvement of nitric oxide in cardioprotective effect of endothelin receptor antagonist during ischemia-reperfusion

The interaction between the cardioprotective effect of endothelin (ET) receptor blockade and nitric oxide (NO) during ischemia-reperfusion injury was investigated. Anesthetized pigs were subjected to 45 (protocol 1) or 30 min (protocol 2) coronary artery ligation and 4 h reperfusion. In protocol 1, five groups were given vehicle, the ET(A) receptor antagonist LU-135252 (LU), the NO synthase (NOS) inhibitor N(G)-nitro-L-arginine (L-NNA), L-NNA in combination with LU, or L-NNA in combination with the NO precursor L-arginine (L-Arg) and LU intravenously before ischemia. In protocol 2, two groups were given vehicle or L-NNA. In protocol 1, the infarct size (IS) was 79 +/- 5% of the area at risk in the vehicle group and 93 +/- 2% in the L-NNA group. LU reduced the IS to 43 +/- 7% (P < 0.001). The cardioprotective effect of LU was abolished in the presence of L-NNA (IS 76 +/- 6%), whereas addition of L-Arg restored its cardioprotective effect (IS 56 +/- 2%; P < 0.05 vs. vehicle and L-NNA + LU groups). In protocol 2, the IS was 49 +/- 6% in the vehicle group and 32 +/- 4% in the L-NNA group (P = not significant). Myocardial ET-like immunoreactivity (ET-LI) increased in the vehicle group of protocol 1. ET-LI in the ischemic-reperfused myocardium was lower in the groups given LU (P < 0.01) and L-NNA + L-Arg + LU (P < 0.05) but not in the group given L-NNA + LU compared with the vehicle group. These results suggest that the cardioprotective effect of the ET(A) receptor antagonist is mediated via a mechanism related to NO.     

The GADD45A (1506T>C) Polymorphism Is Associated with Ovarian Cancer Susceptibility and Prognosis

GADD45A (growth arrest and DNA damage 45 A) is the first stress-inducible gene identified to be a target of p53. However, no studies to date have assessed variants of the GADD45 gene and their potential relationship to tumor susceptibility. We investigated the association of the GADD45A (1506T>C) polymorphism with ovarian cancer development in 258 ovarian cancer patients and 332 age-matched healthy women as controls using sequence analysis. We found a statistically significant difference in the GADD45A (1506T>C) genotype distributions between the case and control groups (TT vs. TC vs. CC, P = 0.0021) and found that variant 1506T>C was significantly associated with an increased risk of ovarian cancer (P<0.001, OR = 1.71, 95% CI [1.28–2.29]). We observed a statistically significant effect between tumor histology (P = 0.032) and CA125 status (P = 0.021). Carrying the C allele (TC+CC) was associated with an increased risk of positive CA125 (OR = 3.20, 95% CI [1.15–8.71). Carrying the T allele (TT+TC) showed a significant correlation with both higher GADD45A mRNA expression and longer ovarian cancer RFS (relapse-free survival) and OS (overall survival). We are the first group to demonstrate that the GADD45A (1506T>C) polymorphism is associated with ovarian cancer susceptibility and prognosis. These data suggest that GADD45A (1506T>C) is a new tumor susceptibility gene and could be a useful molecular marker for assessing ovarian cancer risk and for predicting ovarian cancer patient prognosis.     

A serological marker of the N-terminal neoepitope generated during LOXL2 maturation is elevated in patients with cancer or idiopathic pulmonary fibrosis

ObjectivesLysyl oxidase like 2 (LOXL2) is associated with poor prognosis in idiopathic pulmonary disease (IPF) and cancer. We developed an Enzyme-linked immunosorbent assay (ELISA) targeting the LOXL2 neo-epitope generated through the release of the signal peptide during LOXL2 maturation.Design and methodsAn ELISA targeting the N-terminal site of the human LOXL2 was developed including technical optimization and validation steps. Serum LOXL2 was measured in patients with breast, colorectal, lung, ovarian, pancreatic and prostate cancer, melanoma, IPF and in healthy controls (n = 16).ResultsA technically robust and specific assay was developed. LOXL2 was detectable in serum from healthy controls and showed reactivity towards recombinant LOXL2. Compared to controls, LOXL2 levels were significantly (p < 0.001–0.05) elevated in serum from patients with breast, colerectal, lung, ovarian and pancreatic cancer (mean range: 49–84 ng/mL), but not in prostate cancer (mean: 36 ng/mL) and malignant melanoma patients (41 ng/mL). Serum LOXL2 was elevated in IPF patients compared to healthy controls (mean: 76.5 vs 46.8 ng/mL; p > 0.001)ConclusionsA specific ELISA towards the N-terminal neo-epitope site in LOXL2 was developed which detected significantly elevated serum levels from patients with above-mentioned cancer types or IPF compared to healthy controls.     

Multiple miscarriages are associated with the risk of ovarian cancer: results from the European Prospective Investigation into Cancer and Nutrition

While the risk of ovarian cancer clearly reduces with each full-term pregnancy, the effect of incomplete pregnancies is unclear. We investigated whether incomplete pregnancies (miscarriages and induced abortions) are associated with risk of epithelial ovarian cancer. This observational study was carried out in female participants of the European Prospective Investigation into Cancer and Nutrition (EPIC). A total of 274,442 women were followed from 1992 until 2010. The baseline questionnaire elicited information on miscarriages and induced abortions, reproductive history, and lifestyle-related factors. During a median follow-up of 11.5 years, 1,035 women were diagnosed with incident epithelial ovarian cancer. Despite the lack of an overall association (ever vs. never), risk of ovarian cancer was higher among women with multiple incomplete pregnancies (HR(≥4vs.0): 1.74, 95% CI: 1.20-2.70; number of cases in this category: n?=?23). This association was particularly evident for multiple miscarriages (HR(≥4vs.0): 1.99, 95% CI: 1.06-3.73; number of cases in this category: n?=?10), with no significant association for multiple induced abortions (HR(≥4vs.0): 1.46, 95% CI: 0.68-3.14; number of cases in this category: n?=?7). Our findings suggest that multiple miscarriages are associated with an increased risk of epithelial ovarian cancer, possibly through a shared cluster of etiological factors or a common underlying pathology. These findings should be interpreted with caution as this is the first study to show this association and given the small number of cases in the highest exposure categories.     

Sensory lateralization in pain subjective perception for noxious heat stimulus

The aim of the present study was to identify and characterize hemispheric lateralization for pain intensity perception. A sample of 351 healthy volunteers was tested by the immersion of the right hand for 10 s followed by the same test for the left hand (RL group; n = 199) or in a random sequence (RND group; n = 152) into a water bath (48 degrees C, 15 s). Pain intensity was self-reported by the Visual Analogue Scale (VAS). The motor hemispherical Lateralization Index (LI) was obtained by the Edinburgh Inventory. Gender, hand skin fold, interstimulus time and menstrual cycle data in case of female subjects were recorded. The sample, 60.7% females and 39.3% males, 20.4 +/- 0.18 (mean +/- SEM) years old, showed 92.1% right-handed subjects. Left hand VAS was significantly higher than right hand VAS for RL (7.24 +/- 1.31 vs 6.74 +/- 1.52; p < 0.01) and RND (7.24 +/- 0.82 vs 6.73 +/- 1.25; p < 0.01) both for right- and left-handed subjects. A low but significant correlation for VAS scores and LI was found (r = 0.14; p < 0.05 or r = 0.18; p < 0.05, for left or right hand, respectively). Skin fold was statistically similar in both hands (p > 0.05) being highly correlated with each other (r = 0.68; p < 0.05). Pain subjective perception was not correlated to interstimulus time (r = -0.01; p > 0.05). Females showed significantly higher values than males for both left and right hand VAS scores. Periovulatory phase VAS value was significantly higher than luteal phase VAS only for the right hand test (7.57 +/- 0.20 vs 6.47 +/- 0.33; p < 0.01). The results of the present study suggest a lateralization of pain intensity perception to the right hemisphere not correlated with the motor hemispheric lateralization.     

Oral health awareness in Croatian and Italian urban adolescents

Purpose of this study was to investigate and compare differences in oral health awareness between Croatian and Italian urban adolescents. The sample consisted of primary school last grade students aged between 13 and 15 years, 300 children from Zagreb (Croatia) and 298 children from Bari (Italy). Oral health awareness was evaluated using a self-administered standardized questionnaire. Self-perception of oral health proved to be different between the two groups (p < 0.001). The Croatians reported that their oral health was "excellent" or "very-good" more often than the Italians (68.6% vs. 50.2%). The reasons given for visiting a dentist were different (p < 0.001). The Italians cleaned their teeth more often than the Croatians ("two or more times a day", 83.1% vs. 72.2%, p < 0.003). Wooden toothpicks were preferred by the Croatians (p < 0.001), while floss was preferred by the Italians (p = 0.03). The awareness regarding the use of fluoridated toothpaste was higher in the Italian group (95.6% vs. 72.5%, p < 0.001). The Croatians were consuming sweetened foods more often than the Italians (p < 0.001). Croatian adolescents reported more indicators of a lower level of oral health awareness than the Italians, while on the contrary Croatians had higher esteem of their oral health. Defining national preventive strategies is essential for improving adolescents' attitudes toward oral health in both countries, particularly in Croatia.     

Pre-counseling education materials for BRCA testing: does tailoring make a difference?

Although tailored print materials (TPMs) have been assessed for a variety of behavioral targets, their effectiveness as decision aids for genetic testing had not been evaluated at the time this study began. We compared TPMs and non-tailored print material (NPMs) that included similar content about genetic testing for breast and ovarian cancer susceptibility. TPMs were prepared especially for an individual based on information from and about her. We mailed baseline surveys to 461 women referred by physicians or identified through a tumor registry. All had personal and family histories of breast and/or ovarian cancer and, on the basis of these histories, an estimated > or =10% probability of carrying a mutation in the breast/ovarian cancer genes BRCA1 or BRCA2. The 325 (70%) who responded were randomly assigned to receive TPM or NPM. Followup surveys, mailed 2 weeks following receipt of print materials, were returned by 262 women (81% of baseline responders). Participants were predominately white (94%) and well-educated (50% college graduates). The mean age was 49 years. At follow-up, TPM recipients exhibited significantly greater improvement in percent of correct responses for the 13-item true/false measure of knowledge (24% increase for TPM vs. 16% for NPM; p < 0.0001) and significantly less over-estimation of risk of being a mutation carrier (40% TPM group overestimated vs. 70% NPM; p < 0.0001). Anxiety did not differ significantly between groups. Reactions to materials differed on two items: "seemed to be prepared just for me" (76% TPM vs. 52% NPM; p < 0.001) and "told me what I wanted to know about BRCA1 and 2 testing" (98% TPM vs. 91% NPM; p < 0.05). TPMs showed an advantage in increasing knowledge and enhancing accuracy of perceived risk. Both are critical components of informed decision making.     

Difference in the relation between infarct and occluded bed in pentobarbital-anesthetized and conscious dogs

The relationship between myocardial infarct size (IS) and occluded bed size (OBS) in pentobarbital-anesthetized (A, n = 16) and conscious (C, n = 20) dog models were compared. IS and OBS (postmortem coronary arteriography) were measured by computerized planimetry of weighed left ventricular (LV) rings 7 days after permanent left anterior descending (LAD, n = 19) or circumflex (LC, n = 17) coronary artery occlusion. For both A and C groups, IS was directly related to OBS (p less than 0.001) and no infarcts developed for small occluded beds. For either LAD or LC subgroups, infarcts were larger in A than C dogs (49 +/- 18 vs. 30 +/- 19% OBS, p less than 0.025), with greater slope of the linear regression between IS and OBS (p less than 0.001) and less epicardial sparing on topographic mapping (p less than 0.05). Although postocclusion mean arterial and left atrial pressures were similar in A and C groups, heart rates were greater in the A dogs, both pre- (125 vs. 88 beats/min, p less than 0.001) and post-occlusion (151 vs. 108 beats/min, p less than 0.001). Endocardial flows (radioactive microspheres) in infarct centers and margins were less in A than C dogs. Also, endocardial/epicardial (endo/epi) flow ratios in all regions were less in A than C dogs, both pre- and post-occlusion. Increasing heart rate in 10 other C dogs with LAD occlusion to that of the A group (151 beats/min) by right ventricular pacing resulted in larger infarcts with greater slope of the linear regression and less endo/epi flow ratios, as in the A group.(ABSTRACT TRUNCATED AT 250 WORDS)     

miR-181a通过调控上皮间质转化过程影响卵巢癌细胞的迁移和侵袭

目的:探讨mi R-181a在卵巢癌细胞中的表达及对卵巢癌细胞的迁移和侵袭的影响和可能机制。方法:采用细胞免疫荧光检测卵巢癌细胞中抗波形蛋白和E-钙粘蛋白的表达,Western blotting检测mi R-181a对抗波形蛋白和E-钙粘蛋白的表达情况的调控;划痕愈合实验检测mi R-181a对卵巢癌细胞迁移能力的影响;Transwell侵袭实验检测mi R-181a对卵巢癌细胞侵袭能力的影响。结果:增加mi R-181a的表达后,EMT过程中的相关蛋白激活水平下调,mi R-181a一定程度上可以抑制卵巢癌细胞的EMT过程;过表达mi R-181a后可以明显影响Vimentin[D1(4.58±0.85)vs(0.29±0.02),P0.05;D5(4.16±0.79)vs(0.29±0.02),P0.05]和E-Cadherin[D1(4.75±0.41) vs.(4.56±0.38),P0.05;D5(2.19±0.18) vs.(4.56±0.38),P0.05]蛋白的表达;COC1细胞的迁移能力随mi R-181a的表达的增高而降低[(19.24±4.31)%vs.(25.95±6.02)%,P0.05;(51.25±8.75)%vs.(73.49±12.54)%,P0.05];过表达mi R-181a后可以一定程度上减弱卵巢癌COC1细胞的侵袭能力[(74.64±8.21)vs(231.98±21.72),P0.05]。结论:mi R-181a通过调控上皮间质转化过程影响卵巢癌细胞的迁移和侵袭行为。     

卵巢上皮癌中ING4 基因启动子的甲基化状态及其临床意义

目的：探讨卵巢上皮癌中ING4 基因启动子的甲基化状态及其临床意义。方法：收集2005 年7 月至2012 年6 月哈尔滨医科 大学附属第一医院行全面分期手术并经病理检查确诊的150 例卵巢上皮癌组织标本,并以同期因子宫肌瘤或子宫腺肌症行子宫 全切除术或次全切除术并经病理检查确诊为正常卵巢组织的150 例标本作为对照组。采用甲基化特异性PCR（MSP）技术检测卵 巢上皮癌组织与正常卵巢组织中ING4 基因启动子的甲基化状态,蛋白印迹法检测ING4 蛋白的表达,并分析ING4 基因启动子 的甲基化状态与卵巢上皮癌临床病例特征的关系。结果：卵巢上皮癌组织中ING4 基因启动子的甲基化阳性率为42.7%（64/150）, 明显高于正常卵巢组织（4%,6/150）,差异有统计学意义（P<0.05）。ING4 基因启动子甲基化阳性的卵巢上皮癌组织中ING4蛋白 表达阴性或弱阳性;ING4 基因启动子甲基化阴性的卵巢上皮癌和正常卵巢组织中ING4 蛋白表达阳性;在64 例ING4 基因启动 子甲基化的卵巢上皮癌组织中,ING4 蛋白表达强度与ING4 基因启动子的甲基化程度呈负相关（r=-0.435,P<0.05）。卵巢上皮癌 组织中,ING4 基因甲基化的阳性率随着手术病理分期和组织学分级的增加而增加（P<0.05）;卵巢透明细胞癌（55.6%,10/18）和卵 巢子宫内膜样癌（59.3%,16/27）中ING4 基因甲基化的阳性率显著高于浆液性囊腺癌（33.9%,20/59）和粘液性囊腺癌（39.1%, 18/46）（P<0.05）;ING4基因启动子的甲基化状态与患者的年龄、有无腹水及淋巴结转移均无显著相关性（P>0.05）。结论：ING4 基 因启动子的甲基化可能促进了其在卵巢上皮癌组织中的表达失活,进而促进了卵巢上皮癌的生长和分化。     

NFKB1 insertion/deletion promoter polymorphism increases the risk of advanced ovarian cancer in a Chinese population

Ovarian cancer is the leading cause of death among all gynecological cancers. This is mainly attributed to its frequent presentation at an advanced stage (International Federation of Gynecology and Obstetrics stage III-IV). Nuclear factor-kappaB (NF-κB) is critically involved in the carcinogenesis and development of ovarian cancer. A functional insertion/deletion polymorphism (-94 ins/del ATTG) in the promoter region of the NFKB1 gene, which encodes the p50 subunit of the NF-κB protein, has been recently identified and shown to increase the susceptibility to many diseases. The purpose of this study was to explore the association between this polymorphism and the risk of advanced ovarian cancer in a Chinese population. A total of 179 advanced ovarian cancer patients and 223 healthy controls were recruited into this study. Genotypes were determined using polymerase chain reaction-capillary electrophoresis method. The insertion increased the risk of advanced ovarian cancer (odds ratio?=?2.111, 95% confidence intervals?=?1.125-3.961, p?=?0.019 for heterozygote insertion, and odds ratio?=?2.656, 95% confidence intervals?=?1.397-5.051, p?=?0.002 for homozygote insertion) compared with homozygote deletion. Similar results were seen in age-adjusted analyses (p?相似文献   

TRPM3表达上调通过调控Beclin1的表达促进卵巢癌细胞自噬

目的:探讨瞬时受体电位离子通道3(TRPM3)和Beclin1在卵巢癌中的表达和对卵巢癌细胞自噬的影响。方法:收集6例正常卵巢组织标本和20例卵巢癌组织标本,应用免疫组织化学染色法检测TRPM3和Beclin1在卵巢癌组织中的表达,采用western blotting法检测TRPM3和Beclin1在正常卵巢上皮细胞Moody和卵巢癌细胞Hey、ES-2中的表达差异。用TRPM3-siRNA瞬时转染细胞Hey和ES-2,通过western blotting法检测TRPM3基因沉默情况及Beclin1、p62和LC3的蛋白表达变化。结果:在正常卵巢组织和卵巢癌组织中,TRPM3的阳性表达率分别为33.3%、80.0%(P0.05),而Beclin1的阳性表达率分别为33.3%、65.0%(P0.05)。与正常卵巢上皮细胞Moody相比,卵巢癌细胞Hey和ES-2中TRPM3、Beclin1蛋白表达水平明显较高(P0.05)。沉默TRPM3基因表达的卵巢癌细胞中Beclin1和LC3蛋白表达与对照组相比明显降低,而p62表达升高(P0.05)。结论:卵巢癌组织和细胞中TRPM3蛋白呈高表达,可能通过调控Beclin1促进卵巢癌细胞的自噬。     

Cytokine gene polymorphisms can alter the effect of Bacillus Calmette-Guérin (BCG) immunotherapy

Various types of cancer are more frequent in men than women, and bladder cancer is one of the most common of these. Intravesical instillation of Bacillus Calmette-Guérin (BCG) after transurethral resection is the most effective treatment for superficial bladder cancers. The main aim of this study was to investigate for possible links between cytokine gene polymorphisms and different outcomes after BCG immunotherapy. Sixty patients who had been diagnosed with transitional cell cancer were investigated. All genotyping experiments were performed using polymerase chain reaction sequence-specific primers and a commercially available kit. The genes investigated were those that code for interleukin (IL)-1alpha, IL-1beta, IL-1R, IL-1RA, IL-4RA, IL-2, IL-4, IL-6, IL-10, IL-12, interferon-gamma (IFN-gamma), transforming growth factor-beta (TGF-beta), and tumor necrosis factor-alpha (TNF-alpha). Analyses of the data identified TGF-beta codon 25 GG (92.85% vs. 64.44%, p=0.04, OR=7.17), IL-4 -1098 GG (16.6% vs. 0.0%, p=0.05, OR=18.33), IL-10 -1082 GG (28.5% vs. 6.8%, p=0.05, OR=5.47), and IL-10 -1082 GCC/GCC (28.57% vs. 4.5%, p=0.025, OR=8.4) polymorphisms as risk factors for progression of bladder cancer.     

Adiponectin and the systemic inflammatory response in weight-losing patients with non-small cell lung cancer

The aim of the present study was to examine the relationship between adiponectin and the systemic inflammatory response in weight-losing patients with non-small cell lung cancer (NSCLC). Measurement of anthropometry, acute phase proteins, interleukin-6, leptin (total and free) and adiponectin were carried out on healthy subjects (n = 13) and non-small cell lung cancer patients with weight loss (n = 20). The groups were age and sex matched. Compared with the controls the cancer group had a lower BMI (p < 0.01), mid-upper arm circumference (p < 0.001), triceps skinfold thickness (p < 0.05) and circulating concentrations of albumin (p < 0.001), haemoglobin (p < 0.05), free and total leptin (p < 0.05) and adiponectin (p < 0.01). In contrast, the cancer group had elevated circulating concentrations of interleukin-6 and C-reactive protein concentrations (p < 0.001). In the cancer group circulating adiponectin concentrations were significantly inversely correlated with both free (rs = -0.675, p = 0.001) and total leptin concentrations (rs = -0.690, p = 0.001). However, neither weight loss, interleukin-6 or C-reactive protein concentrations were correlated with either adiponectin, free or total leptin concentrations in the cancer group. These results suggest that adipokine production is normal and is unlikely to play a major role in the abnormal fat metabolism in weight-losing cancer patients.     

卵巢癌中结缔组织生长因子的表达及其临床病理意义

目的:探讨卵巢癌中结缔组织生长因子(CTGF)的表达及其与卵巢癌临床病理特征的关系。方法:采用免疫组化法检测116例卵巢癌和25例正常卵巢组织中CTGF表达,并分析其与卵巢癌临床病理参数的关系。结果:正常卵巢组织及卵巢癌中CTGF的阳性表达率分别96%(24/25)、65%(75/116),卵巢癌中CTGF的阳性表达率显著低于正常卵巢组织(P0.05)。CTGF的表达降低与卵巢癌患者的淋巴结转移、腹膜内转移及对化疗的敏感性显著相关(P0.05);进一步的多因素Logistic回归分析显示CTGF的表达与卵巢癌腹膜转移(P=0.006,OR=4.185,95%CI=1.447-11.352)和淋巴结转移(P0.001,OR=6.336,95%CI=2.563-15.972)均显著相关。结论:卵巢癌中CTGF的表达缺失,是卵巢癌腹膜转移和淋巴结转移的独立影响因素,可能作为预测卵巢癌转移的参考指标。     

Attenuation of heat shock-induced cardioprotection by treatment with the opiate receptor antagonist naloxone

Whole body hyperthermia induces heat shock proteins (HSPs), which confer cardioprotection. Several opioid receptor subtypes are expressed in the heart and are linked to cardioprotection; however, no one has attempted to link the protection elicited by heat stress (HS) to opioids. Therefore, we investigated the effect of an opiate receptor antagonist, naloxone, on HS-induced cardioprotection. Anesthetized Sprague-Dawley rats were subjected to HS (42 degrees C for 20 min) with and without naloxone pretreatment and were allowed to recover for 48 h. They then underwent 30 min of ischemia followed by 2 h of reperfusion. An acute HS group was given an intravenous bolus of naloxone (3 mg/kg) 10 min before index ischemia. Infarct size (IS), expressed as a percentage of the area at risk (IS/AAR), was determined. The right heart was excised for analysis of HSP content by Western blot. Heat-shocked rats showed significant reductions in IS/AAR versus control (16 +/- 3 vs. 58 +/- 4%, P < 0.001). Pretreatment with naloxone before HS attenuated the protective effects in a dose-dependent fashion, with significant attenuation of protection occurring at 15 mg/kg naloxone versus heat shock (42 +/- 6 vs. 16 +/- 3%, P < 0.001). Acute treatment with naloxone (3 mg/kg) 48 h after recovery from HS also significantly attenuated the delayed protective effect (47 +/- 4 vs. 16 +/- 3%, P < 0.001). No difference was seen in the level of HSP70 induced in the different groups. We conclude that heat shock-induced cardioprotection can be attenuated by naloxone, an opiate receptor antagonist, without reducing the levels of certain HSPs. These results suggest there may be a link between the endogenous release of opioids and HS that mediates cardioprotection.     

Can serum amyloid A or macrophage colony stimulating factor serve as marker of amyloid formation process?

Amyloid formation depends on amyloid precursor production and is influenced by the activity of the underlying disorder and mediated by some proinflammatory cytokines. In this pilot study we tried to find some specific markers that could establish the activity of the disease. We investigated 45 samples of sera and 38 samples of urine from patients (pts) with secondary amyloidosis (AA), primary amyloidosis (AL), systemic autoimmune diseases with renal impairment (Vasc) and healthy controls (Co). Pts with AA had increased plasma levels of TNF alpha (9.97 +/- 4.22 vs. 2.63 +/- 1.34 pg/mL, p < 0.001) and SAA (43.14 +/- 16.0 vs. 3.42 +/- 0.7 ng/mL, p < 0.05) in comparison with Co. Plasma levels of M-CSF in the AA group were significantly increased in comparison with Co (1077.34 +/- 238.6 vs. 137.71 +/- 19.6, pg/mL, p < 0.001) and also in comparison with Vasc (482.24 +/- 86.7 pg/mL, p < 0.05). Urinary excretions of TNF alpha (8.92 +/- 8.1 vs. 0.17 +/- 0.11 microgram/mol creatinine, p < 0.01), sIL-6R (1.39 +/- 1.14 vs. 0.07 +/- 0.05 g/mol creatinine, p < 0.01) and M-CSF (650.2 +/- 153.7 vs. 33.3 +/- 8.6 micrograms/mol creatinine, p < 0.01) in AA were significantly increased in comparison with Co. Pts with AL had increased plasma levels of M-CSF (819.83 +/- 264.2 vs. 137.71 +/- 19.6 pg/mL, p < 0.05) and urinary excretion of M-CSF (865.0 +/- 188.4 vs. 33.3 +/- 8.6 micrograms/mol creatinine, p < 0.01) in comparison with Co. SAA has a low specificity for amyloidosis but is a sensitive acute phase reactant. TNF alpha, a proinflammatory cytokine, may reflect the activity of the underlying diseases in secondary amyloidosis. M-CSF was increased both in plasma and urine in amyloidosis groups and seems to be the most promising (possibly specific) marker of amyloidosis.