基于UPLC/MS技术的乳腺癌血清代谢组学研究

目的：探索乳腺癌患者与健康人血清代谢组学差异,为乳腺癌的鉴定与诊断寻找血清潜在生物标志物,推测乳腺癌的发生机理。方法：利用超高压液相色谱质谱（UPLC/MS）技术建立了乳腺癌血清代谢指纹图谱。分别采用主成分分析（PCA）、偏最小二乘法-判别分析（PLS-DA）和正交校正的偏最小二乘法-判别分析（OPLS-DA）分析病例组和正常对照组之间的代谢差异。结果：通过变量重要性投影,结合质谱同位素分析和数据库检索筛选鉴定出了9个潜在生物标志物,涉及核苷代谢、磷脂代谢、氨基酸代谢、脂肪酸代谢及皮质甾体激素代谢。结论：乳腺癌患者与健康对照人群在血清代谢水平上具有明显差异,差异代谢物的发现有助于为发现乳腺癌诊断的潜在标志物提供依据,推测哪些代谢通路参与了乳腺癌的发生发展,为乳腺癌发病机理研究与个体化治疗提供依据。     

基于UPLC-TOF/MS的小细胞肺癌患者血清代谢组学研究

目的 探索小细胞肺癌患者与健康人血清代谢组学差异,为小细胞肺癌的鉴定与分期寻找血清潜在生物标志物。方法 利用超高效液相色谱-飞行时间质谱联用仪(UPLC-TOF/MS)建立小细胞肺癌血清代谢图谱,采用EZinfo2.0软件进行主成分分析(Principal component analysis,PCA)、正交隐变量投影判别分析(Orthogonal partial least squares discriminant analysis,OPLS-DA)分析病例组和正常对照组之间的代谢差异。通过聚类分析并利用HMDB和METLIN数据库搜索差异物的精确质荷比,对一些具有显著性差异的物质进行初步的成分鉴别。结果 通过质谱分析和数据库检索筛选并鉴定出溶血性磷脂酰胆碱等10种差异性代谢产物,不同分期的小细胞肺癌患者轮廓分析存在甘氨胆酸等10种差异性物质。结论 小细胞肺癌患者与健康对照人群在血清代谢水平上具有明显差异,差异代谢物的发现为小细胞肺癌的鉴定以及分期寻找潜在标志物提供实验依据。     

气相色谱-质谱法在乳腺癌血清代谢组学中的应用分析

目的 筛选可用于乳腺癌早期诊断的特异性肿瘤标志物,为乳腺癌的早期诊断提供新的方法。方法 采集41例乳腺癌女性患者和20例健康人血清样本,应用气相色谱-质谱联用技术开展乳腺癌血清代谢组学研究,建立乳腺癌血清代谢指纹图谱。结果 随机森林算法(RF)能有效区分乳腺癌Ⅱ期组、乳腺癌Ⅲ期组及健康对照组。在乳腺癌组样本和健康组样本中共筛选出4种具有明显差异的代谢物：肉豆蔻酸、棕榈酸、硬脂酸、1-棕榈酸单甘油酯。结论 肉豆蔻酸、棕榈酸、硬脂酸、1-棕榈酸单甘油酯的水平变化可能反映了乳腺癌在发展过程中发生的潜在代谢变化,可引起脂肪酸代谢紊乱,有可能成为潜在的乳腺癌标志物。     

鼻咽癌血清NMR代谢组学研究

目的:应用核磁共振氢谱和PLS-DA分析方法研究鼻咽癌患者血清的代谢组学变化.方法:临床正常人血清20例和鼻咽癌患者血清20例,采用日本电子600MHz超导核磁共振波谱仪进行检测,并运用PLS-DA分析方法,得出鼻咽癌患者血清代谢组的特征,探讨相关生化过程的改变.结果:鼻咽癌患者和正常人血清的代谢组学具有明显差异.鼻咽癌患者血清中的甲硫氨酸、牛磺酸和胆碱类物质的含量升高,而脂类、异亮氨酸、不饱和脂类、氧化三甲胺和糖类降低.结论:通过上述代谢物的变化可以区分正常人和鼻咽癌患者.这种基于核磁共振氢谱和PLS-DA分析的代谢组学方法可以为鼻咽癌的诊断提供可靠的代谢依据.     

基于HPLC/Q-TOF-MS的甲状腺乳头状癌组织代谢组学研究

目的：用组织代谢组学方法,探讨甲状腺乳头状癌（papillary thyroid carcinoma, PTC）组织及癌旁组织的代谢差异,寻找PTC的潜在生物标志物,探索PTC的发病机制与治疗策略。方法：收集2018年10月至2020年2月期间湖南省人民医院乳甲外科手术切除的40例PTC患者的癌组织及癌旁组织标本。利用HPLC-MS技术平台对PTC组织及癌旁组织样本的差异性代谢物进行多维统计分析,寻找与PTC相关的异常代谢通路。结果：经PCA、PLS-DA、OPLS-DA分析得知癌组织和癌旁组织的代谢轮廓具有显著性差异。经OPLS-Loading plot分析,结合VIP>1、FC>2,且P<0.05,筛选出76个潜在差异性代谢物。其中亮氨酸、2-褪黑素、香草酸等33种代谢物在PTC组织中表达上调;3-葡萄糖苷、甘油磷脂、磷脂酰胆碱、乳糖等43代谢物在PTC癌组织中表达下调。寻找到与差异性代谢物相关的13条异常代谢通路,如半胱氨酸与蛋氨酸代谢、与甘油磷脂代谢、嘧啶代谢、半乳糖代谢以及丙氨酸、天冬氨酸和谷氨酸代谢、柠檬酸循环等,这些代谢通路可能参与PTC代谢的病理生理过程。ROC曲线下面积大于0.9的差异性代谢物有5种,分别是庚二酸、糖醇、辛二酸、乳糖和L-丝氨酸。结论：PTC组织中半乳糖代谢和氨基酸代谢发生改变,PTC组织细胞中存在沃伯格效应(Warburg effect)。庚二酸、糖醇、辛二酸、乳糖、L-丝氨酸五种差异性代谢物可以用来区分PTC患者与正常人。     

基于代谢组学预测乳腺癌新辅助化疗疗效及预后的研究进展

乳腺癌是一种异质性疾病,表现为新辅助化疗(neoadjuvant chemotherapy,NAC)疗效差异大、预后差异大。代谢组学可以对体内外肿瘤细胞受刺激或扰动后的内源性小分子代谢物进行动态、全面和定量的检测。现有的研究表明,通过对肿瘤、体液等多种标本的代谢组学检测,可以对乳腺癌进行代谢分型,可以预测治疗效果,可以预测疾病预后。虽然其存在一定的局限性,但代谢组学仍为一种新的研究肿瘤的良好手段。全文就代谢组学应用于预测乳腺癌NAC疗效及预后的研究进展进行综述。     

基于UHPLC-QTOFMS代谢组学技术的胃癌进展代谢标志物研究

目的：运用代谢组学方法研究甘肃省农村地区浅表性胃炎患者和胃癌患者血浆中代谢产物的变化,为胃癌的早期诊断提供新的依据。方法：基于超高效液相色谱 四极杆飞行时间质谱联用技术的代谢组学方法对甘肃农村地区的３０例浅表性胃炎患者和３０例胃癌患者血浆样本进行代谢组学分析,利用主成分分析法和正交偏最小二乘法进行数据分析识别,寻找内源性差异代谢物,探讨浅表性胃炎进展为胃癌的代谢特征。结果：根据多变量统计分 析结果,在正离子模式下筛选出２０种差异代谢物,在负离子模式下筛选出１９种差异代谢物,二十碳五烯酸、白三烯、 花生四烯酸、磷酸胆碱等内源性代谢物含量显著下调,Ｌ-半乳糖-１,４-内酯、５-甲基四氢叶酸等代谢物含量显著上调, 提示胃癌患者体内形成了新的代谢调控网络。胃癌组患者代谢通路改变涉及花生四烯酸代谢、α-亚麻酸代谢、甘油磷脂代谢、精氨酸和脯氨酸代谢、赖氨酸代谢、磷酸戊糖途径、生物素代谢、半乳糖代谢、不饱和脂肪酸生物合成等。 另外,ｎ-６／ｎ-３型多不饱和脂肪酸比例的变化及 Ｌ-半乳糖-１,４-内酯向抗坏血酸的代谢转化途径改变可能与甘肃农村地区胃癌的进展密切相关。结论：胃癌患者血浆中二十碳五烯酸等代谢物或可作为潜在的胃癌进展生物标志物,为肿瘤疾病的发病机制和早期筛查提供参考。     

基于代谢组学的胃癌检测研究进展

摘 要：代谢组学是一种快速有效识别新的癌症生物标志物的方法,已逐渐成为基因组学和蛋白质组学的补充技术。全文通过整理近五年基于代谢组学技术研究胃癌检测的相关研究,归纳胃癌相关特异代谢产物及物质代谢路径,认为今后可从葡萄糖或支链氨基酸代谢等作为切入点来探究胃癌检测的代谢组学相关物质指标。     

纳米铜经口染毒大鼠血清的代谢组学研究

背景与目的:运用代谢组学技术检测纳米铜染毒大鼠的血清代谢成分变化,结合常规血液生化及组织病理学检查,探讨纳米铜短期暴露的损害特征,寻找损害早期的代谢标记物,揭示血液代谢表型变化与纳米铜靶器官损害间的关系.材料与方法:雄性Wistar大鼠30只,分为溶剂对照组(1%HPMC),微米铜(200 ms/kg)组和3个不同纳米铜剂量(50、100和200 mg/kg)组,共5组,每组6只,10 ml/kg经口染毒,每日1次,连续5d,次日麻醉采血制备血清,进行核磁共振和血液生化分析,同时摘取肝肾作组织病理学检查.结果:纳米铜200 mg/kg组大鼠血清丙氨酸氨基转移酶、天冬氨酸氨基转移酶、甘油三酯、总胆红素、总胆汁酸、尿素氮和肌酐均明显升高,伴随碱性磷酸酶和总胆固醇明显降低;肝细胞出现点状坏死,肾小管上皮细胞广泛坏死,管腔内可见蛋白管型和棕黄色结晶物沉积,肾小球亦受累及;50、100 mg/kg剂量组甘油三酯和总胆固醇升高,组织病理检查显示肝脏无明显损害,肾小管上皮细胞出现肿胀;微米铜染毒大鼠仅出现肾小管上皮细胞肿胀.血清代谢组学分析表明50～200 mg/kg纳米铜短期暴露可引起能量代谢紊乱和剂量依赖性的血清甘油三脂、不饱和脂肪酸和磷脂水平升高.结论:相同质量浓度的纳米级铜粉毒性明显大于微米级铜粉,肝脏和肾脏是大鼠纳米铜暴露的潜在靶器官,其损害可能与细胞的能量代谢紊乱有关.     

基于UPLC-Q-Exactive-MS的T1期甲状腺乳头状癌血清非靶向代谢组学研究

目的 使用非靶向血清代谢组学方法,借助超高效液相色谱-四级杆-静电场轨道阱高分辨质谱(UPLC-Q-Exactive-MS)联用平台,探讨T₁期甲状腺乳头状癌(PTC)的分子标志物。方法 使用UPLC-Q-Exactive-MS平台,对比分析2021-09-01-2022-04-15民航总医院耳鼻咽喉头颈外科接受甲状腺切除术的30例PTC患者和30名健康体检者的血清代谢物,使用SIMCAP 14.1进行主成分分析(PCA)、偏最小二乘判别分析(PLS-DA)、正交偏最小二乘判别分析(OPLS-DA)建模,结合经FDR校正的Mann-Whitney-Wilcoxon检验结果和代谢物在2组中的差异倍数等筛选潜在的小分子代谢标志物,并通过二元逻辑回归分析建立联合诊断模型。结果 PCA、PLS-DA、OPLS-DA模型显示T₁期PTC患者与健康体检者的血清代谢物存在显著差异。经OPLS-Loading plot分析,结合差异权重贡献值(VIP)>1,|log2FC|>1,且P<0.05,筛选出50种潜在差异性代谢物。其中D-甘油醛3...     

UPLC/Q-TOF MS/MS技术评价防腐剂硼酸和叠氮化钠对尿液代谢物的影响

目的：在基于超高效液相色谱-四级杆-飞行时间串联质谱（UPLC/Q-TOF MS/MS）的尿液代谢组学研究中,确定硼酸和叠氮化钠两种防腐剂是否适用于尿液样品的处理,并确定其适宜的浓度范围。方法：收集6名健康志愿者的晨尿,等量混匀后分装,根据处理方式不同,分为对照组、叠氮化钠组和硼酸组。对照组样品不添加任何防腐剂,叠氮化钠组在尿液中分别添加终浓度为0.1、1和10 mmol/L的叠氮化钠,硼酸组在尿液中分别添加终浓度为2、20和200 mmol/L的硼酸。应用UPLC/Q-TOF MS/MS技术对样品进行检测,采用主成分分析、偏最小二乘判别分析和正交偏最小二乘判别分析等化学计量学方法进行数据处理,比较不同处理组间的代谢图谱。结果：偏最小二乘判别分析得分图显示,3个浓度的叠氮化钠处理组以及2 mmol/L硼酸处理组与对照组间无明显分离趋势;200 mmol/L硼酸处理组与对照组分离趋势明显,两组尿液代谢物存在差异。结论：0.1~10 mmol/L叠氮化钠和2 mmol/L硼酸适用于基于UPLC/Q-TOF MS/MS技术的尿液代谢组学研究。     

Serum immunoglobulins in women with breast cancer.

Serum immunoglobulins were determined in 120 women with breast cancer. A strong association between age at tumor diagnosis and serum IgA levels was noted. Relative hypoglobulinemia of IgA was associated with appearance of tumors early in life, and was more frequent among women with family history of breast cancer and among those with poor prognosis. A strong association between levels of serum IgG and parity was noted. Parous women and/or those who had good prognosis had higher levels of serum IgG, compared to nulliparous women and those with poor prognosis. The observations indicate that serum immunoglobulins may be useful in the identification of women who have a high breast cancer risk and poor prognosis.     

Serum biomarkers for detection of breast cancers: a prospective study

Using surface-enhanced laser desorption/ionization-time of flight (SELDI-TOF), Li et al. [Clin Chem 48(8): 1296–1304, 2002] identified 3 serum biomarkers, BC1 (4.3 kDa), BC2 (8.1 kDa) and BC3 (8.9 kDa), whose combination significantly detects breast cancer patients from non-cancer controls. This work aimed to validate these biomarkers in an independent prospective study. We screened 89 serum samples including 49 breast cancers at pT1-4N0M0 (n = 23), pT1-4N1-3M0 (n = 17) or pT1-4N0-3M1 (n = 9) stages, 13 benign breast diseases and 27 healthy women. The BC2 biomarker significance was not recovered. However, we found 2 peaks that we named BC1a (4286 Da) and BC1b (4302 Da), that could correspond to Li’s BC1 since they significantly decrease in breast cancers (p < 0.00007 and p < 0.0002, respectively). Similarly, BC3a (8919 Da) and BC3b (8961 Da) are significantly increased in breast cancers (p < 0.02 and p < 0.0002, respectively) and could correspond to the Li’s BC3. For each biomarker we defined stringent (no errors) and flexible (less than 10% errors) cut-off values and tested the power of the combined BC1a/BC1b/BC3a/BC3b stringent and flexible profiles to discriminate breast cancers. They identified 33% and 45% cancers, respectively. Applied to the same series, Ca 15.3 test identified 22% patients. Interestingly, in association with the BC1a/BC1b/BC3a/BC3b profiles, Ca 15.3 improved the number of detected cancers indicating that it is an independent parameter. Collectively, our data partially validate those of Li’s study and confirm that the BC1 and BC3 biomarkers are helpful for breast cancer diagnosis.     

Phase I/II study of gemcitabine in association with vinorelbine for metastatic breast cancer

Background. Gemcitabine (G) and vinorelbine (V) have favorable safety profile and antitumor activity in metastatic breast cancer. To exploit their different mechanism of action and lack of overlapping toxicity, we performed a phase I and II study of G and V in combination. Patients and methods. Fifty-three patients with metastatic breast cancer were treated. In the dose-finding phase, seven cohorts of patients (22 women) received increasing doses to determine the dose limiting toxicity (DLT) and maximum tolerated dose (MTD) of the combination. Patients recruited in the phase II portion of the study (31 women) received the dose level immediately below the one defined as MTD (i.e., G 1200mg/m², V 30mg/m², on day 1 and day 8, every 3 weeks). Results. Dose escalation was discontinued at G 1400mg/m² and V 30mg/m² because of toxic death due to thrombocytopenia and CNS hemorrage. No other limiting toxicities were observed, and tolerability was similar at all dose levels studied in the escalation portion of the study. The main toxicity was granulocytopenia of grade 3/4 in 36 and 48% of the patients on phase I and II respectively, without episodes of neutropenic fever. Thrombocytopenia was uncommon. Other side effects were usually mild to moderate. In 46 evaluable patients, the response rate was 24% (complete response 7%, partial response 17%). Disease stabilization was observed in further 17%. The median duration of response was 12 months (range 5–14) and the median survival was 20 months (range 1 to 45+). Conclusions. G and V, on day 1 and 8 of 3-weekly cycles, can safely be administered to patients with metastatic breast cancer at the dose of 1200 and 30mg/m², respectively. The antitumor activity of G and V in combination was similar to that reported when using either drug as single agent.     

Phase I/II study of capecitabine and vinorelbine in pretreated patients with metastatic breast cancer.

PURPOSE: To define the maximum-tolerated dose (MTD) and to evaluate the dose-limiting toxicities (DLT) of the combination of capecitabine and vinorelbine in patients with metastatic breast cancer who relapse after adjuvant and/or first-line treatment. In addition, we aimed to obtain data on efficacy and safety at the recommended dose. PATIENTS AND METHODS: Patients with measurable metastatic breast cancer after failure of prior chemotherapy (including anthracyclines and/or taxanes) were eligible. Capecitabine was administered with a fixed dose of 1000 mg/m(2) orally twice daily for 2 weeks followed by 1 week rest. One treatment cycle consisted of 6 weeks of treatment containing two treatment periods of capecitabine. Vinorelbine was given intravenously at escalated doses of 25 mg/m(2) (dose level 1) and 30 mg/m(2) (dose level 2) on days 1 and 8, and 22 and 29. RESULTS: Thirty-three patients received a total of 91 cycles of capecitabine and vinorelbine. The median number of administered cycles per patient was three (range one to six). Thirty-one patients were evaluable for toxicity. At dose level 2 four out of seven patients experienced DLTs (nausea/vomiting, febrile neutropenia, grade 4 neutropenia, infection and diarrhea); thus, the MTD was defined. In order to confirm the safety and efficacy, dose level 1 was extended to 24 patients. Two patients [8.3%; 95% confidence interval (CI) 1% to 27%] showed DLTs (hospitalization due to febrile neutropenia and prolonged neutropenia). The main toxicity was neutropenia, which was observed at National Cancer Institute Common Toxicity Criteria grade 3 and 4 in 39% of patients. The overall response rate for capecitabine and vinorelbine was 55% (95% CI 36% to 72.7%), including three patients with a complete remission. The median time to disease progression was 8 months (95% CI 4.3-11.7) with an overall survival of 19.2 months (95% CI 11.3-27.1) based on intention-to-treat analysis. CONCLUSIONS: The combination of capecitabine and vinorelbine can be administered with manageable toxicity and showed significant efficacy for patients with metastatic breast cancer even after failure of a anthracycline- and/or taxane-based therapy.     

The impact of bilateral breast cancer on the prognosis of breast cancer: a comparative study with unilateral breast cancer

BACKGROUND: The clinical significance of bilateral breast cancer is unclear and its influence on prognosis is controversial. We assessed the impact of synchronous and metachronous bilateral breast cancer on the prognosis compared with unilateral breast cancer. METHODS: Between January 1, 1960 and December 31, 2001, 1,214 women were treated for primary operable breast cancers. Thirteen (1.1%) had synchronous bilateral breast cancer; 33 (2.7%) had a metachronous contralateral breast cancer. We compared age at operation, menopausal status, clinical stage, tumor size and histology, lymph node status, hormone receptor status, and use of adjuvant chemotherapy or hormone therapy, and we analyzed the impact of these factors on recurrence and survival in the 46 patients with bilateral breast cancer and the 1,168 patients with unilateral breast cancer. RESULTS: The 5-and 10-year disease-free survival rates, respectively, were 65% and 65% in metachronous cases, 85.7% and 64.3% in synchronous cases, and 77.9% and 72.1% in unilateral cases. There was no significant difference in overall survival among the three groups. On multivariate analysis, metachronous bilaterality, tumor size, lymph node status and adjuvant hormone therapy were each independent risk factors for recurrence, whereas bilaterality of breast cancer did not influence overall survival. CONCLUSIONS: Our data suggest that metachronous bilateral breast cancer is associated with shorter disease-free survival than synchronous bilateral or unilateral breast cancer, although overall survival does not differ among the 3 groups. Patients with metachronous bilateral breast cancer should be followed particularly closely in order to detect recurrence early and maximize quality of life.     

代谢组学在卵巢癌研究中的应用

卵巢癌是女性生殖器常见的三大恶性肿瘤之一,致死率居妇科恶性肿瘤首位。随着分子生物学的发展,肿瘤标志物已经成为恶性肿瘤诊断的常用方法之一。代谢组学是近年来出现的一种新的系统生物学研究手段,在药物代谢分析、恶性肿瘤的早期诊断等方面取得了新进展。本文就代谢组学的概念及发展,研究方法及在卵巢癌早期诊断、分型、分期中的应用进行综述。     

Serum enterolactone concentration is not associated with breast cancer risk in a nested case-control study

The lignan enterolactone produced by the intestinal microflora from dietary precursors has been hypothesized to protect against hormone-dependent cancers. We conducted a nested case-control study to examine the relationship between serum enterolactone concentration and risk of breast cancer. Enterolactone concentrations were measured by time-resolved fluoroimmunoassay in serum collected at 4 independent cross-sectional population surveys from 206 women with breast cancer diagnosed during follow-up (mean 8.0 years) and from 215 controls frequency-matched to cases by study cohort, 5-year age group and study area. Mean serum enterolactone concentration (nmol/l) did not significantly differ between case and control subjects [25.2 (SD 22.2) vs. 24.0 (SD 21.3), respectively]. Odds ratios for breast cancer risk estimated by conditional logistic regression for increasing concentration of enterolactone in quartiles were 1.00 (referent), 1.67 (95% CI 0.95-2.95), 1.71 (95% CI 0.96-3.06) and 1.30 (95% CI 0.73-2.31), and p for trend was 0.48. Our findings do not support the hypothesis that high serum enterolactone concentration is associated with reduced risk of breast cancer.