外周血 KISS1 mRNA对上皮性卵巢癌的诊断价值

目的:通过与CA125比较,探讨外周血KISS1 mRNA在上皮性卵巢癌(epithelial ovarian cancer,EOC)中的诊断价值。方法:采集2010年1月至2014年1月在南通大学第二附属医院住院拟手术的EOC患者40例手术前一天的外周静脉血和正常健康者静脉血,用RT-PCR法和电化学发光法分别检测外周血KISS1 mRNA和CA125的表达。结果:EOC早期组(Ⅰ~Ⅱ期)、晚期组(Ⅲ~Ⅳ期)KISS1 mRNA的表达量均显著高于正常健康者(均P<0.01),而早期与晚期EOC组间比较无显著差异(P>0.05);EOC晚期组CA125的表达量明显高于早期组及正常健康者(均P<0.01),而早期组与正常健康者比较无显著差异(P>0.05)。KISS1 mRNA ROC(receptive operator character curve,ROC)曲线的切值(cutoff)设为0.51、0.72时,阳性预测值(positive predictive value,PPV)分别为0.58和1,阴性预测值(negative predictive value,NPV)分别为0.92和0.7;CA125的cutoff值设为20、100 U/ml时,PPV分别为0.72和1,NPV分别为0.87和0.81。KISS1 mRNA、CA125对EOC均具有中度诊断效能(P=0.34);两者比较,KISS1 mRNA对早期EOC具有较高诊断价值(P=0.018),而CA125对晚期EOC的诊断价值较高(P<0.01)。结论:患者外周血KISS1 mRNA与CA125对EOC具有同等的诊断价值,KISS1 mRNA可作为EOC的一个新的肿瘤标志物;两者的联合检测可提高EOC的诊断效能,特别是早期EOC的诊断率。     

卵巢癌组织及腹腔积液中肿瘤干细胞的分离与培养

目的:从人卵巢癌组织和腹腔积液中分离培养肿瘤干细胞,分析细胞表型和化疗药物敏感性。方法:取卵巢浆腺癌患者的原发肿瘤组织及腹腔积液,通过无血清培养形成悬浮生长的球体。采用FCM及免疫荧光法检测干细胞标志物的表达。体外药敏实验检测获得的球体细胞和分化细胞对紫杉醇和卡铂的药物敏感性。结果:卵巢癌组织及腹腔积液中的卵巢癌细胞经无血清悬浮培养可以形成球体,后者具有CD44+CD24-表型并表达Oct4、Nanog及Sox2等干细胞标志物;腹腔积液及组织来源的球体细胞经血清培养液培养3周后发生分化,CD44+CD24-细胞比例分别下降至47%和3%;分化细胞对紫杉醇和卡铂的敏感性均显著高于球体细胞(P<0.01)。结论:卵巢癌的肿瘤干细胞属于CD44+CD24-细胞,后者表达Oct4等干细胞标志并具有显著耐药性。     

Characterization of the GNMT‐HectH9‐PREX2 tripartite relationship in the pathogenesis of hepatocellular carcinoma

The pathogenesis of hepatocellular carcinoma (HCC) involves many molecular pathways. Glycine N‐methyltransferase (GNMT) is downregulated in almost all HCC and its gene knockout mice developed HCC with high penetrance. We identified PREX2, a novel PTEN inhibitor, as a GNMT‐interacting protein. Such interaction enhanced degradation of PREX2 through an E3 ligase HectH9‐mediated proteasomal ubiquitination pathway. Depletion of GNMT or HectH9 resulted in AKT activation in a PREX2 dependent manner and enhanced cell proliferation. An elevated PREX2 protein expression accompanied by activation of AKT was observed in the liver of Gnmt knockout mice. PREX2 protein expression was upregulated in 54.9% of human HCC samples, while its mRNA level was comparable in tumor and tumor‐adjacent tissue, suggesting a post‐translational alteration of PREX2 expression. Higher level of PREX2 in the tumor tissues was associated with poorer survival. These results reveal a novel mechanism in which GNMT participates in AKT signaling and HCC tumorigenesis by promoting HectH9‐mediated PREX2 degradation.     

进展期胃癌S-1与5-FU基础化疗有效性和安全性Meta分析

目的 S-1被应用于进展期胃癌一线化疗中,其疗效也备受关注.本研究评估S-1基础化疗对比5-氟尿嘧啶(5-Fluorouracil,5-FU)基础化疗方案在进展期胃癌一线化疗中的有效性和安全性.方法 用“胃癌、替吉奥或S-1、5-氟尿嘧啶和随机对照研究”等检索词,在Pubmed、Embase、Cochrane Library、ASCO会议摘要、中国生物医学文献数据库(CBM)、中国期刊全文数据库(CNKI)和中文科技期刊全文数据库(VIP)等检索相关的临床随机对照试验,检索时间截止至2016-10.提取总生存期、无疾病进展生存期、有效率、3～4级不良反应等数据.采用Revman 5.3和STATA 12.0进行数据分析.结果 共纳入18个随机对照研究,3 581例患者.结果显示,S-1基础化疗在总生存期(HR=0.92,95％CI为0.84～1.01,P=0.07)及无疾病进展生存期(HR=0.90,95％CI为0.76～1.07,P=0.25)方面与5-FU基础化疗方案差异无统计学意义,但有更高的有效率,RR=1.46,95％CI为1.22～1.74,P＜0.001.S-1基础化疗方案3～4级中性粒细胞减少(P＜0.001)、白细胞减少(P＜0.001)、血小板减少(P=0.01)、口腔炎(P＜0.001)和脱发(P=0.02)等不良反应较5-FU基础化疗发病率更低,差异有统计学意义.结论 相比于5-FU基础化疗,S-1基础化疗在进展期胃癌一线治疗中均是有效且更安全的化疗方案.     

周围型肝内胆管细胞癌超声造影与增强MRI的对比研究

目的 :比较周围型肝内胆管细胞癌的超声造影与增强磁共振成像(magnetic resonance imaging,MRI)的影像学特点,旨在提高影像学诊断的准确性。方法 :对51例经手术或穿刺病理证实为周围型肝内胆管细胞癌的患者的超声造影和增强MRI表现进行分析和比较。结果 :51例患者共有55个病灶。超声造影显示55个病灶在动脉期均有增强;门脉期有8个呈现为等回声,47个呈现为低回声;延迟期55个病灶均呈现低回声。增强MRI显示55个病灶在动脉期均有增强;门脉期和延迟期延迟强化的病灶有44个,11个病灶呈现为低回声。超声造影显示有42个(76.4%)病灶表现为周边环状增强,13个(23.6%)病灶表现为整体增强,增强过程中有40个(72.7%)病灶表现为"树枝样"向内延伸的增强方式;增强MRI显示40个(72.7%)病灶表现为周边环状强化,15个(27.3%)病灶表现为整体强化。超声造影的特征性表现为动脉期病灶周边环状增强,并呈"树枝样"向内延伸,达峰值时大多数病灶表现为不均匀增强;增强MRI的特征性表现为动脉期周边环状增强,而门脉期和延迟期表现为延迟强化。以"快进快出"、周边环状增强和(或)"树枝样"增强作为周围型肝内胆管细胞癌超声造影的特征,其出现率为70.9%(39/55);而以动脉期周边环状增强和延迟增强作为增强MRI的特征性表现,其出现率为69.1%(38/55)。超声造影与增强MRI的特征性影像出现率差异无统计学意义(P=1.000)。结论 :周围型肝内胆管细胞癌的超声造影和增强MRI均有特征性表现,具有较高的诊断价值。     

Confirmation of genetic variants associated with lethal prostate cancer in a cohort of men from hereditary prostate cancer families

Germline genetic variants have been suggested as prognostic biomarkers for identifying patients at high risk for lethal prostate cancer (PCa). Validation studies have confirmed the association of several single nucleotide polymorphisms (SNPs) with fatal PCa, but whether these variants affect PCa‐specific mortality (PCSM) in patients with an inherited predisposition to PCa, based on familial history, is unknown. For this study, a cohort of 957 PCa patients from 270 hereditary prostate cancer families of European ancestry was genotyped for a panel of 22 PCSM‐associated SNPs. Death certificates were reviewed to confirm cause of death. Mixed‐effect Cox proportional hazards models were used to assess survival according to genotypes, accounting for relatedness and clinicopathological factors. Within this cohort, 98 PCa deaths were confirmed over an average follow‐up period of 12.7 years after diagnosis. Variant allele carriers for three SNPs had significantly altered risk for PCSM [rs635261 at RNASEL, hazard ratio (HR), 0.35, 95% CI, 0.18–0.66; p = 0.002; rs915927 in XRCC1, HR, 1.91, 95% CI, 1.21–3.02; p = 0.009; and rs2494750 at AKT1, HR, 0.45, 95% CI, 0.23–0.90; p = 0.016). These results confirm the association of genetic variation in three genes with PCa lethality in a cohort of men with an inherited susceptibility to the disease and provide validation evidence that germline SNPs provide prognostic information for PCa patients. Development of a panel of germline biomarkers with clinical utility for distinguishing patients at detection who have an increased risk for fatal PCa is warranted.     

反义寡核苷酸对骨肉瘤细胞MTA1基因表达及侵袭性影响的研究

目的:合成人肿瘤转移相关基因(MTAl)的反义脱氧寡核苷酸,观察其转染后对人骨肉瘤MG-63细胞系中MTA1表达及骨肉瘤侵袭性的影响.方法:免疫细胞染色观察人工合成正义、反义及无意义MTA1基因片段转染人骨肉瘤细胞MG-63后,人肿瘤转移相关基因的表达;RT-PCR和蛋白质印迹法检测MTA1基因mRNA和蛋白质表达水平的变化;Boyden小室体外侵袭实验检测转染前后细胞侵袭力的变化.结果:反义寡核苷酸处理骨肉瘤细胞后,MTA1 mRNA的表达下降(吸光度之比为25.09±0.21),与正义链组、无意义链组和空白对照组(35.19±0.17、33.20±0.23和37.17±0.18)相比差异有统计学意义,P<0.05.Boy-den小室体外侵袭实验检测显示,反义寡核苷酸处理后骨肉瘤细胞的透膜能力明显下降.结论:MTA1同骨肉瘤的转移能力密切相关,反义寡核苷酸的转染可阻遏骨肉瘤细胞中MTA1的表达,并因此有可能限制骨肉瘤的侵袭和转移.     

Frequent hypermethylation of RASSF1A and TSLC1, and high viral load of Epstein-Barr Virus DNA in nasopharyngeal carcinoma and matched tumor-adjacent tissues

We examined the promoter hypermethylation of tumor-suppressor genes RASSF1A and TSLC1, quantitated EBV DNA load in nasopharyngeal carcinoma (NPC) tissues (T tissues), and matched tumor-adjacent tissues outside 0.5 cm (P tissues) and outside 1.0 cm (Z tissues) to evaluate the role of promoter hypermethylation of RASSF1A and TSLC1 as well as viral load in the pathogenesis of NPC. Methylation-specific polymerase chain reaction (PCR) for RASSF1A and TSLC1 and quantitative real-time PCR analysis of EBV DNA were performed on matched T, P, and Z tissues (n = 28) as well as chronic nasopharyngitis tissues (n = 8). Hypermethylated RASSF1A was frequently detected in the T (82%) and P tissues (75%), but less frequently in Z tissues (46%). he average quantities of EBV DNA (copies/microg DNA) in matched T, P, and Z tissues were 673,000, 90,000, and 7000. The differences of promoter hypermethylation of RASSF1A and EBV viral load among T, P, and Z tissues were statistically significant, with more frequent methylation and higher viral load detected when tissues examined were nearer to the NPC tissues. Our results suggest that aberrant hypermethylation of RASSF1A and high EBV load might be important events in NPC pathogenesis, and they may be useful molecular diagnostic markers for this cancer.     

卡培他滨用于52例蒽环类和紫杉类治疗失败的转移性乳腺癌

[目的]观察卡培他滨单药应用于蒽环类和紫杉类治疗失败的转移性乳腺癌的近期疗效和不良反应。[方法]对既往接受过蒽环类和紫杉类药物的转移性乳腺癌患者，口服卡培他滨14d，21d为1个周期，每例患者至少进行2个周期的治疗。观察近期疗效和不良反应。[结果]52例患者中无1例完全缓解，部分缓解率（PR）10例（19．2％），病情稳定（SD）26例（50．0％），病情进展（PD）16例（30．8％），临床获益率（CR＋PR＋SD）69．2％。常见的不良反应是手足综合征31例（59．6％），皮肤色素沉着21例（40．4％），恶心呕吐8例（15．4％），腹泻6例（11．5％），骨髓毒性和肝脏毒性较轻。[结论]单药卡培他滨治疗蒽环类和紫杉类失败的转移性乳腺癌仍有一定的疗效，患者耐受性良好。