活性氧在肿瘤顺铂耐药中的作用研究进展

顺铂是目前应用最广泛的抗癌药物之一。作为DNA络合物,顺铂不仅能够通过结合并损伤细胞核DNA,诱导DNA损伤应答,而且能够通过结合线粒体DNA,诱导线粒体功能紊乱,活性氧（ROS）过度产生,最终导致细胞凋亡坏死。ROS是一类化学性质活泼的氧分子衍生物的统称,主要由细胞内线粒体以及NADPH过氧化物酶产生。过量的ROS能够导致核酸、脂质以及蛋白质发生氧化损伤,是顺铂抗肿瘤效应的重要介质。然而,在肿瘤顺铂耐药的进程中,ROS亦发挥了重要作用。因此,本文综述了顺铂耐药肿瘤中ROS的代谢特点和ROS调控顺铂耐药的多种机制,包括上调顺铂转运体表达、增强细胞DNA损伤修复能力、提高核转录因子红系2相关因子2、低氧诱导因子-1α的蛋白水平和转录活性以及调控线粒体动态活动等方面。最后,本文还总结了ROS诱导剂和抗氧化系统抑制剂这两类ROS调节药物的相关研究,希望为提高顺铂疗效,克服肿瘤顺铂耐药提供治疗新思路。     

LncRNA与非小细胞肺癌顺铂耐药的研究进展

<正>在全世界,肺癌是发病率最高的癌症,同时具有较高死亡率。非小细胞肺癌是肺癌的常见病理类型,由于暴露在油烟等环境下,中国女性非小细胞肺癌的发病率较国外高^[1-3],因起病的隐匿性,患者出现症状就医时已是晚期,含顺铂的化疗方案是常见晚期非小细胞肺癌的治疗手段,其耐药性的出现极大的影响了患者的预后。长链非编码RNA是一种长度大于200个核苷酸单位的不具备编码能力的RNA,许多研究发现其在恶性肿瘤的发生、发展^[4-6]甚至对逆转顺铂耐药发挥了一定的作用,本文就近些年来LncRNA与非小细胞肺癌的发生、发展及对顺铂耐药相关的研究做综述,期待可以为非小细胞肺癌的临床诊治及逆转顺铂耐药提供新的思路及突破口。1 Lnc RNA与非小细胞肺癌的发生发展研究发现LncRNA与非小细胞肺癌的发生发展相关,丁梦杰等^[7]的回顾性研究选取了90例NSCLC患者作为研究对象,     

顺铂致严重低钙血症1例

1病例介绍患者女,44岁,农民,因小细胞肺癌近3年,拟化疗于2014年3月5日入院。既往有顺铂过敏可疑。体检：身高155 cm,体质量51 kg,T 36.5℃,R 19次/min,HR 79次/min,BP 123/88mmH g,体力状况评分（Performance status,PS）0分。双侧颈部未触及明显肿大淋巴结,右侧腋下可触及一巨大肿块,直径12cm,左侧腹股沟可触及直径约5 cm肿大淋巴结,质韧,固定,边界清,无压痛;     

T-钙黏蛋白与肝癌肝移植术后肿瘤复发转移的相关性研究

目的 探讨T-钙黏蛋白在肝癌肝移植患者癌组织和癌旁组织中的表达及其与肿瘤复发转移的关系。方法 收集解放军联勤保障部队第九〇〇医院2003年1月-2012年12月行原位肝移植手术的183例肝细胞癌（HCC）患者的临床资料,以符合肝癌肝移植米兰标准,且被切除肝脏的癌旁组织无微小血管侵犯（MVI）的76例患者为研究对象。根据术后随访中肿瘤是否复发转移分为复发转移组和无复发转移组,采用免疫组化法检测T-钙黏蛋白基因,观察其在两组患者癌组织和癌旁组织中的表达,比较术后5年内出现复发转移患者与未出现复发转移患者癌组织中T-钙黏蛋白基因的表达差异。结果 纳入的76例患者5年总复发转移率为21.05%,其中无MVI的患者46例（60.52%）,术后肝癌复发转移率为17.39%;肝癌组织中细胞膜T-钙黏蛋白阳性表达率降低,缺失和异常表达明显升高,占47.83%,而癌旁组织缺失及异常表达较少见,仅占6.52%;两组相比差异具有统计学意义（χ²=19.828,P=0.000）;在出现复发转移的肝癌肝移植患者中癌组织T-钙黏蛋白异常表达（100%）与未复发转移的肝癌肝移植患者的（36.84%）相比差异有显著的统计学意义（χ²=10.565,P=0.001）。结论 T-钙黏蛋白基因在HCC中缺失和异常表达率较高,且与肝移植术后肝细胞癌的复发转移密切相关,有可能作为判断HCC肝移植预后的预测指标和药物干预的靶点。     

miR-539增强顺铂对非小细胞肺癌耐药株的杀伤作用研究

目的 研究miR-539对顺铂诱导的非小细胞肺癌(non-small cell lung cancer,NSCLC)耐药细胞株凋亡的影响.方法 通过荧光定量聚合酶链反应(PCR)检测NSCLC细胞系和人正常支气管上皮细胞16HBE中miR-539表达情况;通过转染miR-539建立miR-539过表达细胞系,观察miR-539过表达对顺铂抑制NSCLC细胞生长率的影响,检测细胞凋亡水平及相关基因的表达.结果 NSCLC细胞系中miR-539表达水平低于人正常支气管上皮细胞(P<0.01).阴性对照组培养48、72、96 h NCI-H460细胞增殖率均明显低于空白对照组,转染miR-539组培养不同时间NCI-H460细胞增殖率均明显低于阴性对照组和空白对照组(P<0.01).阴性对照组和转染miR-539组NCI-H460细胞凋亡率高于空白对照组(P<0.05).阴性对照组和转染miR-539组的Bcl-2、Bcl-xL基因表达量均低于空白对照组,且转染miR-539组低于阴性对照组,而Bim、Bax、P53、Myc基因表达量高于空白对照组,且转染miR-539组高于阴性对照组(P<0.05,P<0.01).结论 miR-539能够增强顺铂对NSCLC耐药细胞株的杀伤作用,可作为NSCLC基因治疗的有效靶点.     

奈达铂联合5-氟脲嘧啶治疗顺铂耐药的晚期食管癌临床疗效观察

目的考察奈达铂联合5-氟脲嘧啶（5-Fu）对顺铂耐药的晚期食管癌的临床疗效及毒副反应。方法奈达铂80 mg.m-2静滴2 h,d1;亚叶酸钙（CF）75 mg·m-2静滴2 h后,5-FU 500 mg·m-2持续静滴6 h以上,d1～5,28 d为一周期。完成2～3个周期后,根据WHO标准评价疗效和毒副反应。结果总有效率（CR＋PR）42.4%,1年生存率29.1%;主要毒副作用为骨髓抑制。结论对顺铂耐药的晚期食管癌患者,奈达铂联合5-Fu治疗是安全有效的方案,值得临床推广。     

竹节参皂苷对顺铂耐药肺癌细胞敏感性的影响及机制研究

目的 研究竹节参皂苷对顺铂耐药肺癌细胞敏感性的影响及其潜在作用机制。方法 通过递增顺铂浓度诱导肺癌A549细胞耐药,测定细胞存活率,SRB显色法测定药物敏感性,Annexin V-FITC/PI双染测定细胞凋亡,ELISA测定Caspase 3和Caspase 8水平,Western blotting测定细胞中相关蛋白表达情况。结果 经长期诱导后,A549细胞对顺铂的耐受性增加14.79倍,且较稳定。与顺铂组比较,联合应用竹节参皂苷和顺铂可显著增加A549细胞对顺铂的敏感性,增加细胞内Caspase 3和Caspase 8水平,促进细胞凋亡,同时抑制MDR1、P-gp、Bcl-2和p-Akt等蛋白表达,促进Bax蛋白表达。结论 竹节参皂苷可增加顺铂耐药细胞株对顺铂的敏感性,增强顺铂的作用效果,其作用机制可能与抑制耐药相关蛋白MDR1、P-gp和Akt活性相关。     

T-钙黏蛋白与某些皮肤病的研究进展

钙黏素是Ca依赖性跨膜的细胞间黏附分子，通过在细胞膜表面表达钙粘素配体介导同型细胞间及细胞与基质间的黏附，参与桥粒的构成，维持组织结构的完整，在细胞识别信号转导，细胞的连接和生长分化、组织发育、器官形成等过程中发挥重要的作用。T-钙黏蛋白（即T-cadherin，又称CDH13或H—cadherin）是近来被发现的一种新型非常独特的钙黏附蛋白分子，研究发现T-钙黏蛋白的表达与某些皮肤病的发病机制有关，现综述如下。     

重组人血管内皮抑素联合顺铂治疗鼠B16恶性黑色素瘤移植模型的实验研究

目的 探讨重组人内皮抑素(恩度)联合顺铂化疗对恶性黑色素瘤的作用.方法 建立B16恶性黑色素瘤的裸鼠移植模型20只,随机分为恩度组、顺铂组、恩度联合顺铂组(联合组)和对照组.治疗后处死裸鼠,计算肿瘤体积抑瘤率,用免疫组化法测微血管密度(MVD)及瘤体血管内皮生长因子(VEGF)的表达.结果 恩度组、顺铂组和恩度联合顺铂组(联合组)的肿瘤抑制率分别为17.52%、45.72%和59.40%,联合组的肿瘤生长明显受到抑制(P＜0.05).免疫组化结果显示,联合组MVD低于其他组(P＜0.05);恩度组、顺铂组和联合组的VEGF表达均明显低于对照组(P＜0.05).结论 恩度联合顺铂有协同抗肿瘤作用,能有效抑制恶性黑色素瘤的生长.     

苦参碱介导上皮间质转化逆转肺癌A549顺铂耐药株活性研究

目的 探讨苦参碱(MT)逆转肺癌A549顺铂耐药株活性中的作用及分子机制.方法 对数生长期人肺腺癌耐顺铂细胞株A549/DDP细胞,用不同浓度MT(1,4,16,32μmol·L-1)处理,采用噻唑蓝(MTT)法在24 h检测细胞增殖抑制率.将细胞分为空白对照组及MT组(16μmol·L-1),光学显微镜、Transw...     

Inhibitory activity of Bifidobacterium adolescent combined with cisplatin on melanoma in mice and its mechanism

The aim of this study is to explore inhibitory activity of Bifidobacterium adolescent combined with cisplatin on the growth of melanoma (B16) in mice and the underlying mechanism. C57 mice were inoculated with B16 cancer cells to construct mouse model of melanoma and treated with bifidobacterium adolescent combined with cisplatin. Ratios of inhibitory activity on the growth of melanoma (B16) were calculated. Pathology changes of the tumor were observed by HE staining. B16 cell cycles were examined on a flow cytometer. Lymphocyte proliferation was measured with MTT assay and the T-cell subset was measured by double marked fluorescence. When bifidobacterium of 10¹⁰ cfu/L was injected, the ratio of inhibitory activity on the growth of melanoma (B16) reached 54%, which was similar to that of cisplatin group. The ratio of inhibitory activity reached 74.45% when the mice were treated by bifidobacterium combined with cisplatin. HE staining shows that bifidobacterium inhibited B16 cell proliferation and enhanced the cisplatins killing activity on B16 cells. The results of flow cytometry demonstrated that B16 cell proliferation was arrested at G₁ stage after treatment with bifidobacterium. The B16 cell proliferation was arrested at S stage after treatment with cisplatin. The CD4+ percentage increased and the difference was significant compared with the normal group after treatment with bifidobacterium, indicating that T-cell immune activity was enhanced. Treatment with bifidobacterium combined with cisplatin can enhance the inhibitory activity on the growth of melanoma (B16) of cisplatin. The mechanism of the inhibitory activity on B16 cell proliferation is correlated with the enhanced immune activity in mice. __________ Translated from Journal of the Fouth Military Medical University, 2007, 28(21): 1947–1950 [译自: 第四军医大学学报]     

诺斯卡品逆转人卵巢癌裸鼠移植瘤对顺铂耐药的作用及机制

目的：探讨微管抑制剂诺斯卡品（ NOS ）逆转人卵巢癌细胞株 SKOV3/DDP 裸鼠皮下移植瘤对顺铂（ DDP）耐药的作用及其机制。方法建立人卵巢癌裸鼠移植瘤模型,分为对照组、DDP组、NOS组、NOS与DDP联合组,每组6只。观察各组裸鼠饮食和精神状态,测量裸鼠的体重及移植瘤的体积,并绘制肿瘤生长曲线。用网搓法制备单细胞悬液,采用流式细胞术检测移植瘤细胞周期和细胞凋亡率的变化,细胞中X链锁凋亡抑制蛋白（ XIAP ）、半胱氨酸天冬氨酸蛋白酶（Caspase-3）、B细胞淋巴瘤/白血病-2（Bcl-2）和生存素（Survivin）蛋白表达。结果联合组裸鼠移植瘤体积明显小于对照组和其他用药组（ P ＜0ⅱ．05）。与对照组和DDP组比较,联合组皮下移植瘤细胞凋亡率显著增加（ P ＜0．05）,G2/M期细胞比例增多（ P ＜0．05）,细胞中XIAP、Bcl-2和Survivin蛋白表达降低（ P ＜0．05）, Caspase-3蛋白表达升高（ P ＜0．05）。结论 NOS增加了移植瘤细胞对DDP的敏感性,逆转其对DDP的多药耐药,其机制可能与XIAP、Bcl-2和Survivin蛋白表达的下调,Caspase-3蛋白表达的上调有关。     

Cell context-dependent activities of parthenolide in primary and metastatic melanoma cells

Background and purpose:

Growing evidence implicates NF-κB as an important contributor to metastasis and increased chemoresistance of melanoma. Here, we report the effects of parthenolide on either untreated, cisplatin- or TNFα-treated melanoma cell lines A375, 1205Lu and WM793, exhibiting different levels of constitutive NF-κB activity.

Experimental approach:

Electrophoretic mobility shift assay was used to assess changes in NF-κB activity, and real-time PCR to evaluate expression of NF-κB-regulated genes. Cell cycle arrest and apoptosis were assessed by flow cytometry. Cell death was also visualized by fluorescence microscopy. Migration was determined by scratch assay and invasiveness by Matrigel assay.

Key results:

Parthenolide suppressed both constitutive and induced NF-κB activity in melanoma cells. This was accompanied by down-regulation of cancer-related genes, with NF-κB-binding sites in their promoters, including: Bcl-X_L, survivin, cyclin D1, interleukin 8 and matrix metalloproteinase 9. When the various effects of 6 µM parthenolide were compared, apoptosis associated with loss of mitochondrial membrane potential was most efficiently induced in 1205Lu cells, cell cycle arrest in G₀/G₁ phase was observed in WM793 cells, and high metastatic potential was markedly reduced in A375 cells. These findings not only reflected differences between melanoma cell lines in basal expression of NF-κB-regulated genes, but also suggested other parthenolide targets involved in cell cycle progression, migration, invasiveness and survival.

Conclusions:

Inhibition of constitutive and therapeutically induced NF-κB pathway by parthenolide might be useful in the treatment of melanoma, although the diversity of changes induced in melanoma cells with different genetic backgrounds indicate context-dependent poly-pharmacological properties of this compound.     

顺铂联合吉西他滨与联合多西紫杉醇治疗晚期非小细胞肺癌疗效观察

目的 探讨顺铂联合吉西他滨与联合多西紫杉醇治疗晚期非小细胞肺癌(NSCLC)的疗效、生存率及不良反应.方法 我院60例NSCLC患者随机分为2组,每组30例,1组给予顺铂联合吉西他滨方案(GP组),另1组给予顺铂联合多西紫杉醇方案(TP组),观察2组有效率、生存率及不良反应.结果 GP组的PR为53.3%,TP组的PR...     

丹皮酚协同顺铂抑制人食管癌Eca-109细胞的增殖

目的 探讨丹皮酚单药及联合顺铂应用对人食管癌Eca-109细胞的增殖抑制作用.方法 以不同浓度的丹皮酚、顺铂和两药联用处理人食管癌Eca-109细胞,采用MTT法测定药物对体外培养的人食管癌Eca-109细胞增殖的抑制作用,应用两药相互作用指数(coefficient of drug interaction,CDI)进行联合用药分析.结果 丹皮酚(7.81～250 mg·L-1)和顺铂(0.078～5 mg·L-1)单独应用均对人食管癌Eca-109细胞有抑制作用,呈现剂量效应关系.丹皮酚与顺铂联合应用时,丹皮酚能明显提高顺铂对Eca-109细胞的增殖抑制作用,有显著的协同杀伤作用.结论 丹皮酚与顺铂联合应用具有明显的协同抗食管癌Eca-109细胞增殖的作用.     

Synergistic effect of combining paeonol and cisplatin on apoptotic induction of human hepatoma cell lines

Aim： To investigate whether paeonol （Pae） has synergistic effects with cisplatin （CDDP） on the growth-inhibition and apoptosis-induction of human hepatoma cell lines HepG2 and SMMC-7721. Methods： The cytotoxic effect of drugs was measured by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyl-tetrazolium bromide assay. The coefficient of drug interaction was used to analyze the nature of drug interactions. Morphological changes were observed by acridine orange fluorescence staining. Cell cycle and the apoptosis rate were detected by flow cytometry. Bcl-2 and Bax expression were assayed by immunohistochemical staining. Results： Pae or CDDP had antiproliferative effect on the 2 cell lines in a dose-dependent manner, with different sensitivities to drugs. More interestingly, a synergistic inhibitory effect on the viability of the 2 cell lines was observed after treatment with a combination of Pae （15.63, 31.25, and 62.5 mg/L） with various concentrations of CDDP. Further study showed typical morphological changes of apoptosis if the cells were exposed to the two agents for 24 h. The apoptotic rate of the cells with combination treatment was significantly higher than that of cells treated with Pae or CDDP alone. The expression of Bcl-2 decreased and that of Bax increased in the treated groups, especially in the combination group, with the ratio of Bcl-2/Bax decreasing correspondingly. Additionally, a combination of Pae with CDDP resulted in a stronger S phase arrest, compared with Pae or CDDP alone. Conclusion： Pae, in combination with CDDP, had a significantly synergistic growth-inhibitory and apoptosis-inducing effect on the 2 human hepatoma cell lines, which may be useful in hepatoma treatment.     

多西他赛联合顺铂治疗晚期非小细胞肺癌26例

目的:探讨多西他赛联合顺铂4 wk治疗晚期非小细胞肺癌方案的疗效和安全性。方法:经病理组织学或细胞学确诊的ⅢB期或Ⅳ期非小细胞肺癌病人26例,年龄在35-78 a,ECOG PS评分为0-2分;d 1,8 多西他赛37．5 mg·m-2,d 1-3顺铂75-100 mg·m-2,分3 d,均予静脉滴注。每4 wk重复,2个周期后评价疗效与不良反应,并随访生存期。结果:24例可评价疗效病人中,部分缓解(PR)8例,完全缓解(CR)1例,总有效率为38％。ECOG PS评分0-1分病人有效率明显高于ECOG PS评分2分病人。中位生存期为11 mo,1年生存率为50％。Ⅲ-Ⅳ度白细胞减少的发生率为20％。结论:多西他赛联合顺铂4 wk方案治疗晚期非小细胞肺癌安全且有效。     

Phase II trial of cisplatin and etoposide in patients with metastatic melanoma

Summary Fourteen patients with metastatic melanoma were treated with cisplatin and etoposide by bolus intravenous infusion daily for 5 consecutive days each month. All patients were evaluable for toxicity and twelve for response. Eight patients were treated with cisplatin 20 mg/m² and etoposide 100 mg/m² daily. Because of excessive myelosuppression, the daily dose of etoposide was reduced to 75 mg/m² in the remaining six patients. There were no major responses among 12 evaluable patients (major response rate 24% with 95% confidence). The median time to progression was one month. One patient with a liver metastasis had a minor response lasting 6 + months. The combination of cisplatin and etoposide in these doses and schedule lacked sufficient clinical efficacy in the treatment of metastatic melanoma.     

艾洛替尼耐药肝癌HepG2细胞系的建立及其耐药机制

目的体外建立艾洛替尼(erlotinib)耐药的人肝癌细胞系HepG2/erlotinib,鉴定其生物学特性,并对其耐药机制进行初步探讨。方法逐步递增艾洛替尼并最终给予艾洛替尼50μmol·L-1连续冲击HepG2细胞12个月,建立HepG2/erlotinib。磺酰罗丹明B法检测HepG2/erlotinib耐药倍数和耐药谱;测定细胞生长曲线并计算细胞增殖时间;流式细胞术检测细胞周期;RT-PCR和Western印迹法检测HepG2/erlotinib细胞乳腺癌耐药蛋白(BCRP)基因和蛋白的表达;流式细胞术检测细胞表面BCRP蛋白表达。结果经过12个月艾洛替尼诱导的HepG2耐药细胞,艾洛替尼的IC50值为(72.3±6.1)μmol·L-1,艾洛替尼对正常HepG2细胞的IC50值为(10.6±0.3)μmol·L-1,耐药倍数为6.8±0.7,表明HepG2/erlotinib为艾洛替尼耐药细胞系。HepG2/erlotinib对顺铂、多柔比星、米托蒽醌和拓扑替康的IC50均大于正常HepG2细胞的IC50,表明产生交叉耐药性。HepG2细胞和HepG2/erlotinib细胞的群体倍增时间分别为(20.7±3.3)h和(26.7±1.2)h,耐药细胞倍增时间延长。与HepG2细胞相比,HepG2/erlotinib的S期细胞比例明显增高(P<0.05),G0/G1期细胞比例明显降低(P<0.01);HepG2/erlotinib细胞中BCRP基因和BCRP蛋白表达显著升高(P<0.05)。结论建立的HepG2/erlotinib具有耐药细胞的特征和生物学特性,其耐药机制与BCRP蛋白表达增加有关。     

大蒜素联合顺铂对人宫颈癌细胞增殖的抑制作用

目的探讨大蒜素联合顺铂对人宫颈癌HeLa细胞增殖的抑制作用并探讨其机制。方法取对数生长期HeLa细胞,设对照组、大蒜素（50 μg/mL）组、顺铂（5 μg/mL）组、联合给药组（大蒜素50 μg/mL+顺铂5 μg/mL）。药物干预48 h后,通过MTT法检测细胞增殖抑制率,运用CompuSyn软件计算大蒜素与顺铂联合指数（CI）,流式细胞术检测细胞周期分布和细胞凋亡水平,Western blotting法检测Cyclin D1、CDK2、P27、Cleaved Caspase-3、Bcl-2、Bax蛋白表达。结果与大蒜素组和顺铂组比较,联合给药组HeLa细胞增殖抑制率显著升高（P<0.05、0.01）,CI为0.69（提示大蒜素与顺铂具有中度协同效应）。与对照组比较,大蒜素组和顺铂组G₀/G₁期细胞比例和细胞凋亡率显著升高（P<0.05、0.01）,Cyclin D1、CDK2、Bcl-2表达显著下调且P27、Cleaved Caspase-3、Bax表达显著上调（P<0.05、0.01）,Bax/Bcl-2比值显著升高（P<0.01）。与大蒜素组和顺铂组比较,联合给药组G₀/G₁期细胞比例和细胞凋亡率显著升高（P<0.05、0.01）,Cyclin D1表达显著下调且P27、Cleaved Caspase-3、Bax表达显著上调（P<0.05、0.01）,Bax/Bcl-2值显著升高（P<0.01）。结论大蒜素联合顺铂具有协同抑制人宫颈癌HeLa细胞增殖的作用,可能与调节细胞周期和凋亡相关蛋白表达,进而阻滞细胞周期进程并促进细胞凋亡有关。