NP与GP方案治疗晚期非小细胞肺癌68例疗效分析

为了评价NP和GP方案治疗晚期非小细胞肺癌的疗效和不良反应。将1999年12月2日～2004年5月2日收治的68例非小细胞肺癌（non—small cell lung cancer，NSCLC）患者随机分为两组，分别应用NP和GP方案治疗。NP方案：长春瑞滨（NVB）25mg／m^2，d1、d8；顺铂（DDP）50mg，d3～d5。GP方案：健择（Gemcitabine）1000mg／m^2，d1、d8；DDP50mg，d3～d5，两种方案均21d为1个周期，至少治疗2个周期。结果为NP组35例，无CR，PR17例（48．6％），SD13例（37．1％），PD5例（14．3％），总有效率为48．6％（17／35），临床受益率85．7％（30／35）。GP组33例，CR1例（3．0％），PR14例（42．4％），SD13例（39．4％），PD5例（15．2％），总有效率为45．5％（15／33），临床受益率84．8％（28／33）。NP组和GP组中住进展时间分别为3．2和3．3个月，初治优于复治（NP组60％vs 33％，GP组52．6％ vs 35．7％）。荆量限制性毒性主要为骨髓抑制，NP组和GP组白细胞及血小板下降的发生率分别为80％、22．9％和51．5％、51．5％。NP组静脉炎及胃肠道反应较GP组重（31．4％ vs6．1％和57．1％vs45．5％）。初步研究结果提示，NP和GP方案治疗晚期NSCLC均安全有效，疗效相当，不良反应均可耐受。     

慢性淋巴细胞性白血病患者miR-9-3甲基化异常及其意义

目的本研究旨在探讨microRNA-9-3(miR-9-3)在慢性淋巴细胞性白血病骨髓细胞中的甲基化异常及其意义。方法采用甲基化特异性聚合酶链反应(MSP)技术检测8例正常骨髓组织、78例新确诊的慢性淋巴细胞性白血病患者和7种白血病细胞株甲基化水平。结果正常对照组miR-9-3呈未甲基化状态;7种白血病细胞株中有5种呈未甲基化状态(71.4%);78例慢性淋巴细胞性白血病患者中65例呈miR-9-3甲基化,MSP阳性率为83%。5-氮杂-2'-脱氧胞苷(5-Aza Dc)处理白血病细胞株后I83-E95和WAC3CD5+细胞株miR-9-3呈未甲基化状态。结论慢性淋巴细胞性白血病患者存在miR-9-3表达受抑,可能与其基因甲基化异常有关。而miR-9-3甲基化在激活慢性淋巴细胞性白血病患者NF-κB1信号通路的作用值得进一步研究。     

Deoxycytidine kinase promotes the migration and invasion of fibroblast-like synoviocytes from rheumatoid arthritis patients

Rheumatoid arthritis (RA) is a complex, multi-system disease whose primary site of inflammatory tissue damage is the joint. The increasing evidences indicate that activated RA fibroblast-like synoviocytes (FLS) play a critical role in the development of pannus by migrating into cartilage and bone. Furthermore FLS and T cells can activate each other in vitro and in vivo, which is crucial for the progress of RA. Deoxycytidine kinase (DCK) has been linked to peripheral T cell homeostatic proliferation and survival, which is very important for RA. Yet, the function of DCK in FLS is still unknown. Here, we present a story that DCK could regulate the migration and invasion of FLS through AKT pathway in RA patients. Moreover, DCK seems to be the upstream of AKT and FAK, and AKT inhibitor exerted the similar effect on FLS motility. In summary, our study characterized the new role of DCK in human primary FLS cells, and figured out the possible pathway DCK involved in, and these findings might propose DCK as a novel target for controlling joint destruction of RA.     

去甲基化剂5-氮杂-2''''-脱氧胞苷对胃癌细胞生物学行为的影响

目的:探讨去甲基化剂5氮杂2’脱氧胞苷(5AzaCdR)对胃癌细胞生物学行为的影响。方法:使用浓度5×106mol/L和105mol/L的5AzaCdR处理胃癌细胞株AGS、SGC7901、MKN28及MKN45。通过MTT、平板克隆实验观察处理前、后细胞的生长活性,RTPCR检测处理前、后抑癌基因RASSF1AmRNA表达的改变,并应用流式细胞仪进行细胞周期和凋亡率的分析。结果:4株胃癌细胞经不同浓度之5AzaCdR处理后,与对照组相比,生长速度出现不同程度减慢;实验组AGS细胞较对照组细胞克隆形成率显著降低(32.4%、28.5%比57.0%,P<0.01);5AzaCdR处理后,3株无RASSF1AmRNA表达的细胞(AGS、MKN28和SGC7901)均检出基因重新表达;处理前后4株胃癌细胞无明显G1期、G2/M期改变,AGS细胞凋亡率由0.6%增至18.6%和20.2%,MKN28细胞亦出现凋亡率由1.85%增至3.85%和6.61%。结论:去甲基化剂5AzaCdR对胃癌细胞生长周期无显著影响,可能系通过诱导抑癌基因再表达或凋亡等途径,间接抑制胃癌细胞的生长。     

吉西他滨联合卡培他滨治疗一线耐药乳腺癌的临床观察

为了观察吉西他滨(GEM)联合卡培他滨治疗蒽环类和(或)紫杉类耐药性晚期乳腺癌的疗效和毒副反应,2003-01-2006-03应用GEM联合卡培他滨治疗蒽环类和(或)紫杉类药物耐药的晚期乳腺癌38例,每例治疗2～6个周期,评价疗效并记录毒副反应。38例患者中,CR4例(10.5%),PR12例(31.6%),SD14例(36.8%),PD8例(21.1%),总有效率为42.1%(16/38),临床获益率为78.9%(30/38),中位疾病进展时间为8.5个月(2～59个月),1、2年生存率分别为60.5%和31.6%。主要毒副反应为骨髓抑制、胃肠道反应和手足综合征,Ⅲ～Ⅳ度白细胞减少发生率较低,手足综合征和消化道反应以Ⅰ、Ⅱ度为主。初步研究结果提示,GEM联合卡培他滨对蒽环类物和(或)紫杉类耐药的转移性乳腺癌有较好疗效,且毒副反应可以耐受。     

Deoxycytidine kinase and cN-II nucleotidase expression in blast cells predict survival in acute myeloid leukaemia patients treated with cytarabine

Summary. The cytotoxic activity of cytarabine (ara-C) in leukaemic blasts depends on activating enzymes such as deoxycytidine kinase (dCK) and inactivating enzymes such as the 5'-nucleotidases. We have analysed dCK and 'high-Km' 5'-nucleotidase (cN-II) mRNA expression by the quantitative real-time polymerase chain reaction at diagnosis in leukaemic blasts from 115 acute myeloid leukaemia (AML) patients treated with ara-C. The prognostic value of these parameters as well as that of the cN-II/dCK ratio was determined. In univariate analyses: (1) low levels of dCK, high levels of cN-II and a high cN-II/dCK ratio predicted shorter disease-free survival (DFS); (2) low levels of dCK and cN-II/dCK ratio also predicted shorter overall survival (OS). In a multivariate analysis taking into account other clinical and laboratory variables: (1) high cN-II expression, a high cN-II/dCK ratio, age ≥ 60 years and an unfavourable karyotype were independent prognostic factors for DFS; and (2) a high cN-II/dCK ratio, age ≥ 60 years and an unfavourable karyotype predicted shorter OS. Age, karyotype and cN-II/dCK ratio were used to define a prognostic score that permitted the identification of high- and low-risk groups. Our results suggest that dCK and cN-II mRNA expression in leukaemic blasts at diagnosis is correlated with clinical outcome and may play a functional role in the resistance to ara-C in patients with AML.     

In vitro susceptibility of CD4+ and CD8+ T cell subsets to fludarabine

Administration of the adenosine analogue fludarabine (FLU) in vivo induces a profound and prolonged T lymphopenia which mainly affects CD4(+) cells. To better understand the mechanistic basis underlying this preferential depletion, we analyzed the in vitro susceptibility of T cell subsets to FLU-induced apoptosis. Contrasting with observations in vivo, our results showed that treatment of peripheral blood mononuclear cells with FLU induced a higher level of apoptosis in CD8(+) than in CD4(+) T lymphocytes. This increased sensitivity of CD8(+) T cells to FLU was observed in samples from both, healthy donors and B cell chronic lymphocytic leukemia patients, and resulted in higher CD4:CD8 ratios in FLU-treated than in untreated cultures (P<0.01). Expression of factors involved in FLU transport and metabolism was then evaluated by quantitative real time-PCR in normal T cell subsets. It was found that mRNA levels of human equilibrative nucleoside transporter-1 nucleoside transporter were higher whereas deoxycytidine kinase and IMP/GMP selective 5'-nucleotidase mRNA levels were lower in CD4(+) cells. However the dCK/cN-II ratio was 2-fold greater in CD8(+) than in CD4(+) T lymphocytes, which could account for the higher apoptosis levels observed in the CD8(+) subset. These results favor the view that decreased CD4:CD8 ratios in FLU-treated patients should be attributed to differences in cell recovery and/or homing between T cell subsets.     

Down-regulation of deoxycytidine kinase in human leukemic cell lines resistant to cladribine and clofarabine and increased ribonucleotide reductase activity contributes to fludarabine resistance

Mechanisms of acquired resistance to three purine analogues, 2-chloro-2'-deoxyadenosine (cladribine, CdA), 9-beta-D-arabinofuranosyl-2-fluoroadenine (fludarabine, Fara-A), and 2-chloro-2'-arabino-fluoro-2'-deoxyadenosine (clofarabine, CAFdA) were investigated in a human T-lymphoblastic leukemia cell line (CCRF-CEM). These analogues are pro-drugs and must be activated by deoxycytidine kinase (dCK). The CdA and CAFdA resistant cell lines exhibited increased resistance to the other nucleoside analogues activated by dCK. This was also the case for the Fara-A resistant cells, except that they were sensitive to CAFdA and guanosine analogues. The CdA and CAFdA resistant cells displayed a deficiency in dCK activity (to <5%) while the Fara-A resistant cells showed only a minor reduction of dCK activity (20% reduction). The activity of high K(m) 5'-nucleotidase (5'-NT) (cN-II) using IMP as substrate, was 2-fold elevated in the resistant cell lines. The amount of the small subunit R2 of ribonucleotide reductase (RR) was higher in the Fara-A resistant cells, which translated into a higher RR activity, while CdA and CAFdA cells had decreased activity compared to the parental cells. Expression of the recently identified RR subunit, p53R2 full-size protein, in CAFdA cells was low compared to parental cells, but a protein of lower molecular weight was detected in CdA and CAFdA cells. Co-incubation of Fara-A with the RR inhibitor 3,4-dihydroxybenzohydroxamic acid (didox) enhanced cytotoxicity in the Fara-A resistant cells by a factors of 20. Exposure of the cells to the nucleoside analogues studied here also caused structural and numerical instability of the chromosomes; the most profound changes were recorded for CAFdA cells, as demonstrated by SKY and CGH analysis. We conclude that down-regulation of dCK in cells resistant to CdA and CAFdA and increased activity of RR in cells resistant to Fara-A contribute to resistance.     

5-AZA-2'-deoxycytidine (5-AZA-CdR): a demethylating agent affecting development and reproductive capacity

The objective was to evaluate the effects of 5-AZA-2'-deoxycytidine (5-AZA-CdR) on postnatal development and reproductive capacity. Pregnant mice were administered 1 mg kg-1 5-AZA-CdR at gestation day 10. The body weights of F1 control and treated (in uterine-exposed) pups were recorded. To evaluate the reproductive capacity, 5-AZA-CdR F1 males and females were mated with control mice. The presence of plugs and the number of pregnancies were recorded. The 5-AZA-CdR F1 male mice were killed. Total body, testes and epididymis weights were recorded. Spermatid head counting, histological analyses and serum testosterone levels were performed. Body weights of 5-AZA-CdR F1 mice were statistically lower than controls (P < 0.01), with the females more strongly affected (P < 0.05). Male mating capacity appeared to be more adversely affected. Mating of 5-AZA-CdR F1 males with control females resulted in a lower pregnancy rate compared with control mating groups (P < 0.01). Gross testicular and epididymis weights were lower in 5-AZA-CdR F1 mice (P < 0.01). However, testicular and epididymis weights in these mice were higher than controls when correlated to body weight (P < 0.01). In 5-AZA-CdR F1 male mice, all measured reproductive parameters, including total number of spermatid heads per testis, are significantly lower (P < 0.01) than the controls except for the number of spermatid heads per milligram of testis.