rh-Apo2L联合长春瑞滨对诱导肺鳞癌细胞凋亡作用及机制的研究

目的 晚期肺鳞癌治疗方式相对有限.Ⅲ期临床试验提示,rh-Apo2L有望为其提供一种新颖、有效的治疗方法.本研究旨在观察rh-Apo2L、长春瑞滨及两药联合对人肺鳞癌细胞SK-MES-1的抑制增殖和促进凋亡作用,并探讨其机制.方法 将不同质量浓度的rh-Apo2L、长春瑞滨干预人肺鳞癌SK-MES-1细胞,采用CCK8法检测细胞增殖;Annexin Ⅴ-FITC/PI流式细胞术检测细胞凋亡;蛋白质印迹法检测细胞凋亡相关蛋白DR4、DR5和Caspase-3的表达.结果 通过CCK8检测发现,rh-Apo2L对人肺鳞癌细胞SK-MES-1的体外增殖具有抑制作用,24和48 h的IC50分别为28.57和8.97 μg/mL;长春瑞滨对人肺鳞癌细胞SK-MES-1的体外增殖具有抑制作用,24和48 h的IC50分别为27.30和7.87 μg/mL.其中,1μg/mL rh-Apo2L组、1 μg/mL长春瑞滨组和两药联合组(rh-Apo2L和长春瑞滨质量浓度均为1 μg/mL)的24 h细胞增殖抑制率分别为(15.03±1.54)％、(21.88±2.75)％和(65.11±4.09)％;0.1 μg/mL rh-Apo2L组、0.5 μg/mL长春瑞滨组和两药联合组(rh-Apo2L质量浓度为0.1μg/mL,长春瑞滨为0.5 μg/mL)的48 h细胞增殖抑制率分别为(19.01±1.12)％、(19.97±1.23)％和(84.81±3.99)％;24和48 h联合组两药相互作用指数(q值)分别为1.94和2.40.细胞凋亡实验结果显示,1 μg/mL rh-Ap02L组、1μg/mL长春瑞滨组和两药联合组(rh-Ap02L和长春瑞滨质量浓度均为1 μg/mL)干预细胞24 h的凋亡率分别为(21.76±3.13)％、(37.31±2.21)％和(74.88±3.63)％;与单药组相比较,两药联合组对SK-MES-1细胞增殖的抑制作用明显增强(P＜0.001),两药联合干预24 h的凋亡指数显著升高,P＜0.001.蛋白印迹法检测结果发现,rh-Ap02L联合长春瑞滨在上调DR4(P=0.026)和DR5(P=0.001)表达方面具有交互作用,在增加Caspase-3活性片段表达亦具有交互作用,P=0.011.结论 rh-Apo2L与长春瑞滨均能协同抑制人肺鳞癌SK-MES-1细胞增殖并诱导细胞凋亡,其机制可能与上调SK-MES-1细胞表面的DR4、DR5蛋白表达和活化Caspase-3相关.     

活体自发光成像监测长春瑞滨纳米胶束抗乳腺癌转移机制研究

  目的  乳腺肿瘤细胞的转移是乳腺癌致死的主要原因。淋巴转移作为血道和淋巴两大转移途径之一,在乳腺癌转移过程中至关重要。针对乳腺癌转移的特点,本研究组设计了具有淋巴富集特性的纳米载药系统,活体动态监测其对原位乳腺肿瘤生长以及肺脏和淋巴器官转移的抑制效果,旨在为临床用药提供科学依据。  方法  采用活体自发光成像实时动态地监测了聚乙二醇化磷脂（PEG-PE）包载的长春瑞滨纳米胶束抗乳腺癌体内转移的过程。  结果  与未包载的游离长春瑞滨（Free Vin）相比,PEG-PE包载的长春瑞滨胶束（NanoVin）增强了长春瑞滨的抗原位瘤活性,显著抑制了乳腺肿瘤细胞的淋巴及肺脏转移。  结论  通过静脉给药达到了血道转移和淋巴转移两条途径的双重清扫,为靶向药物设计提供了新思路。活体光学成像技术可动态监测肿瘤的转移,为临床应用影像技术检测药物疗效提供必要的技术手段。      

长春瑞滨联合依维莫司对乳腺癌细胞增殖、凋亡的影响及其机制

目的 探讨长春瑞滨联合依维莫司对乳腺癌细胞增殖、凋亡的影响及其机制.方法 以乳腺癌细胞MCF-7为研究对象,采用长春瑞滨、依维莫司干预细胞,检测长春瑞滨、依维莫司作用细胞48 h的半数抑制浓度(IC50),根据IC50设置长春瑞滨组、依维莫司组及联合组的用药浓度,以不加药物处理的细胞为对照组.MTT法检测细胞增殖活性,流式细胞术检测细胞凋亡率,Western blot检测细胞中磷脂酰肌醇-3激酶(PI3K)、磷酸化PI3K(p-PI3K)、丝氨酸/苏氨酸蛋白激酶1(AKT1)、磷酸化AKT1(p-AKT1)蛋白的表达情况.建立乳腺癌裸鼠转移瘤模型,腹腔注射长春瑞滨(2 mg/kg,qd)或依维莫司(2 mg/kg,q3d),联合组注射长春瑞滨(2 mg/kg,qd)+依维莫司(2 mg/kg,q3d),计算肿瘤体积和重量.结果 随着长春瑞滨或依维莫司药物作用浓度的增加,细胞的存活率逐渐下降,且呈一定的浓度依赖性.联合组的细胞存活率低于长春瑞滨组和依维莫司组(P﹤0.05).长春瑞滨组、依维莫司组的细胞凋亡率均高于对照组,联合组的细胞凋亡率高于长春瑞滨组和依维莫司组,差异均有统计学意义(P﹤0.05).长春瑞滨组、依维莫司组、联合组的肿瘤体积和重量均低于对照组,联合组的肿瘤体积和重量均低于长春瑞滨组和依维莫司组,差异均有统计学意义(P﹤0.05).长春瑞滨组和联合组细胞的p-PI3K、p-AKT1蛋白表达水平均低于对照组,差异均有统计学意义(P﹤0.05);长春瑞滨组细胞的p-PI3K、p-AKT1蛋白表达水平均低于依维莫司组,高于联合组,差异均有统计学意义(P﹤0.05).结论 长春瑞滨和依维莫司在抑制细胞增殖、促进细胞凋亡、抑制肿瘤生长时具有协同作用,主要是通过抑制PI3K/AKT1信号通路的激活发挥作用.     

长春瑞滨联合西妥昔单抗抑制血管生成作用的实验研究

目的 探讨长春瑞滨联合西妥昔单抗对体内外血管生成的抑制作用.方法 以人肺腺癌细胞株A549为对照,采用四甲基偶氮唑蓝(MTT)法,观察长春瑞滨联合西妥昔单抗对人脐静脉内皮细胞(HUVEC)增殖能力的影响;应用Transwell小室模型、体外小管形成实验及流式细胞术,分别观察长春瑞滨联合西妥昔单抗对HUVEC迁移、小管形成及细胞凋亡的影响;应用鸡胚绒毛尿囊膜(CAM)模型,观察药物对体内CAM血管生成的抑制作用.结果 0.1、0.4和0.8 ng/ml长春瑞滨对HUVEC增殖的抑制率分别为25.8%、39.2%和54.0%,0.25 μg/ml西妥昔单抗对HUVEC增殖的抑制率为19.7%.0.1 ng/ml长春瑞滨+0.25 μg/ml西妥昔单抗和0.4 ng/ml长春瑞滨+0.25 μg/ml西妥昔单抗对HUVEC增殖的抑制率分别为29.5%和46.4%,联合用药有次加作用;0.8 ng/ml长春瑞滨+0.25 μg/ml西妥昔单抗对HUVEC增殖的抑制率为64.6%,联合用药有协同作用.0.1、0.4和0.8 ng/ml长春瑞滨分别联合0.25 μg/ml西妥昔单抗具有抑制HUVEC迁移的作用,抗HUVEC迁移率分别为51.9%、68.2%和95.0%,联合应用有协同作用.0.1 ng/ml长春瑞滨+0.25 μg/ml西妥昔单抗或0.4 ng/ml长春瑞滨+0.25 μg/ml西妥昔单抗对抑制HUVEC小管形成有次加或协同作用,抗HUVEC小管形成率分别为38.8%和57.7%;0.8 ng/ml长春瑞滨+0.25 μg/ml西妥昔单抗对抑制HUVEC小管形成有协同作用,抗HUVEC小管形成率为78.9%.0.8 ng/ml长春瑞滨+0.25 μg/ml西妥昔单抗诱导HUVEC的凋亡率为59.9%,有协同作用.长春瑞滨联合西妥昔单抗具有协同抑制CAM血管生成的作用.结论 小剂量长春瑞滨和西妥昔单抗在体内外具有抗血管生成作用,联合用药具有次加或协同作用.     

新型汉防己甲素纳米微球的体外抗肿瘤作用

目的:全面考察我们前期工作中采用mPEG-PCL自行合成的汉防己甲素(Tetradrine,Tet)载药纳米微球的体外抗肿瘤活性,并对其机理进行探讨,并评价mPEG-PCL作为药物载体的生物相容性.方法:通过开环聚合法,制备mPEG-PCL两嵌段聚合物作为载体, 采用乳化-挥发法,制备负载Tet的纳米微球.以MTT法,评价该纳米微球在体外对于胃癌细胞株MKN-28、BGC-823的抗肿瘤活性,并与汉防己甲素裸药相比较.在MKN-28、BGC-823及LO2细胞株上验证空白微球的毒性.采用荧光标记粒子细胞摄取实验,探讨Tet纳米微球抗肿瘤作用不同于裸药的机制.结果:Tet载药微球在作用48h后对于体外培养的肿瘤细胞生长的抑制率与裸药相似,空白微球在极高浓度时,对于肿瘤细胞系及人肝细胞株LO2均无明显毒性.荧光标记粒子的细胞摄取实验证实,微球可通过胞吞作用进入细胞,在细胞内释放药物,这是微球不同于裸药的抗肿瘤机制.结论:Tet微球具有良好的抗肿瘤活性,载体无毒,具有良好的生物相容性.其抗肿瘤作用的机制可能和细胞对于纳米粒子的直接摄取有关,因此可能提高汉防己甲素的体内抗肿瘤效果.     

TRAIL联合吉西他滨对肺癌NCI-H460细胞增殖和凋亡的影响

目的:观察吉西他滨(gemcitabine)联合TNF相关凋亡诱导配体(TNF-related apoptosis inducing ligand,TRAIL)对人肺癌细胞NCI-H460增殖和凋亡的影响。方法:MTT法检测NCI-H460细胞的增殖,流式细胞术检测NCI-H460细胞的凋亡。结果:吉西他滨和TRAIL单用或联用均可浓度(0.001~10μg/ml)和时间(24、48、72 h)依赖性抑制NCI-H460细胞的增殖,两药联用比单用时抑制率更高,3组药物作用72 h时对肿瘤的抑制率分别为14.9%~77.5%、23.4%~74.8%、22.3%~80.5%;作用72 h时两药单用和联用对细胞增殖抑制的IC50分别为0.085 3、0.098 2、0.043 5μg/ml。两药联用对NCI-H460细胞增殖的抑制效果还与用药顺序以及两药比例有关,吉西他滨作用24 h后再加TRAIL比TRAIL作用24 h后再加吉西他滨对NCI-H460的抑制效果更好,吉西他滨与TRAIL联用的比例为1∶0.3时对NCI-H460细胞增殖的抑制率最大。吉西他滨和TRAIL联用时NCI-H460细胞的凋亡率显著高于两药单用的凋亡率(49.04%vs29.33%、25.69%,P<0.01)。结论:吉西他滨与TRAIL联用可协同抑制肺癌细胞NCI-H460的增殖、促进细胞凋亡。     

地西他滨联合三氧化二砷对NB4细胞凋亡作用的研究

 目的　研究去甲基化制剂地西他滨（DAC）单用或联合三氧化二砷（As2O3）对NB4细胞凋亡的作用机制。方法　将不同浓度的DAC、As2O3以及两药联合作用于NB4细胞,不加药为对照组,采用四甲基偶氮唑蓝（MTT）法检测细胞增殖抑制作用,流式细胞术检测细胞凋亡。结果　DAC与As2O3单药对NB4细胞的抑制作用呈浓度时间依赖性（DAC 1 μmol／L作用24、48、72 h的抑制率分别为12.18 %、22.72 %、35.54 %;DAC 2 μmol／L作用24、48、72 h的抑制率分别增高为22.14 %、31.18 %、45.21 %;As2O3 0.5 μmol／L作用24、48、72 h的抑制率分别21.09 %、32.43 %、44.93 %;As2O3 1.0 μmol／L作用24、48、72 h的抑制率分别增高为31.69 %、41.12 %及54.27 %）,两药联合抑制作用较单药明显（DAC 1 μmol／L+As2O3 0.5 μmol／L作目的　研究去甲基化制剂地西他滨（DAC）单用或联合三氧化二砷（As2O3）对NB4细胞凋亡的作用机制。方法　将不同浓度的DAC、As2O3以及两药联合作用于NB4细胞,不加药为对照组,采用四甲基偶氮唑蓝（MTT）法检测细胞增殖抑制作用,流式细胞术检测细胞凋亡。结果　DAC与As2O3单药对NB4细胞的抑制作用呈浓度时间依赖性（DAC 1 μmol／L作用24、48、72 h的抑制率分别为12.18 %、22.72 %、35.54 %;DAC 2 μmol／L作用24、48、72 h的抑制率分别增高为22.14 %、31.18 %、45.21 %;As2O3 0.5 μmol／L作用24、48、72 h的抑制率分别21.09 %、32.43 %、44.93 %;As2O3 1.0 μmol／L作用24、48、72 h的抑制率分别增高为31.69 %、41.12 %及54.27 %）,两药联合抑制作用较单药明显（DAC 1 μmol／L+As2O3 0.5 μmol／L作用24、48、72 h抑制率分别为42.10 %、48.75 %、60.78 %）（P＜0.05）,各浓度组与对照组比较差异均有统计学意义（P值均＜0.05）;As2O3 1 μmol／L作用于NB4细胞株48 h可见5.8 %的细胞凋亡,联合组增高为17.3 %。结论　DAC能显著抑制NB4细胞的增殖并诱导其凋亡,DAC联合As2O3对NB4细胞增殖抑制及诱导凋亡有协同作用。用24、48、72 h抑制率分别为42.10 %、48.75 %、60.78 %）（P＜0.05）,各浓度组与对照组比较差异均有统计学意义（P值均＜0.05）;As2O3 1 μmol／L作用于NB4细胞株48 h可见5.8 %的细胞凋亡,联合组增高为17.3 %。结论　DAC能显著抑制NB4细胞的增殖并诱导其凋亡,DAC联合As2O3对NB4细胞增殖抑制及诱导凋亡有协同作用。     

比卡鲁胺联合紫杉醇对雄激素受体阳性三阴性乳腺癌MDA-MB-231细胞的增殖抑制作用

目的探讨比卡鲁胺(BIC)联合化疗药物紫杉醇(PTX)对雄激素受体(AR)阳性的三阴性乳腺癌MDA-MB-231细胞的增殖抑制作用及可能的作用机制。 方法采用CCK-8试剂盒观察不同浓度的BIC (0.1、1.0、10.0 μmol/L)和PTX(0.1、1.0、10.0、100.0、1 000.0、10 000.0 nmol/L)以单药及不同联合给药方式处理后,对MDA-MB-231细胞增殖的抑制作用。细胞增殖抑制率比较采用单因素方差分析。组间两两比较采用LSD法。选取10 nmol/L PTX及10 nmol/L DMSO分别处理MDA-MB-231细胞样品(各3个)72 h,采用生物信息学方法分析样品的相关基因表达芯片数据,采用校正t检验筛选出差异基因。 结果使用不同浓度的BIC分别处理MDA-MB-231细胞24、48、72 h后,各组MDA-MB-231细胞增殖抑制率在不同时间点差异均有统计学意义(F＝4.124、8.189、4.139, P＝0.037、0.004、0.032)。BIC 10.0 μmol/L组MDA-MB-231细胞增殖抑制率在48 h最高,为(12.9 ± 5.5)%。不同浓度的PTX分别处理MDA-MB-231细胞24、48、72 h后,不同浓度组MDA-MB-231细胞增殖抑制率在不同时间点差异均有统计学意义(F＝8.407、47.432、14.907, P均<0.001)。PTX在48 h时对MDA-MB-231细胞的半数抑制浓度(IC₅₀)为5 380.0 nmol/L。5 000.0 nmol/L PTX单药或联合不同浓度(0.1、1.0、10.0 μmol/L)的BIC同时处理MDA-MB-231细胞48 h后,5 000.0 nmol/L PTX单药处理组与3个实验组中细胞增殖抑制率分别为(53.2±2.7)%、(53.2±3.1)%、(51.7±3.4)%、(51.0±2.3)%,组间差异无统计学意义(F＝0.831,P＝0.492)。采用5 000.0 nmol/L PTX和10.0 μmol/L BIC以不同的序贯方式联合给药处理MDA-MB-231细胞(PTX 24 h ＋BIC 24 h组、BIC 24 h ＋PTX 24 h组、PTX 48 h ＋BIC 24 h组、BIC 48 h＋PTX 24 h组),并用5 000.0 nmol/L PTX(PTX 48 h组)和10.0 μmol/L BIC(BIC 48 h组)单药处理及同时联合给药(PTX 48 h＋ BIC 48 h组)分别处理MDA-MB-231细胞后,各组间细胞增殖抑制率差异有统计学意义(F＝241.466,P<0.001)。其中,两两比较结果显示,PTX 24 h ＋BIC 24 h组细胞增殖抑制率为(72.9±1.9)%,高于BIC 24 h ＋PTX 24 h组的(42.9±1.7)%(P<0.001),PTX 48 h组的(60.9±3.7)%(P<0.001)和PTX 48 h ＋BIC 48 h组的(60.3±4.1)%(P<0.001)。PTX处理组中有EGR1、FST、FOS、IL8、IL6、RPL27A及CA2 7个基因的表达量与DMSO处理组比较,差异均有统计学意义(t＝18.647、10.336、10.098、9.683、9.408、9.050、8.001,P均<0.050)。 结论通过先PTX再BIC的序贯联合给药方式较单药及其他联合给药方式能够更有效抑制AR阳性三阴性乳腺癌细胞MDA-MB-231的增殖,两者间可能存在协同作用。     

长春瑞滨化疗所致静脉炎的防治及护理

长春瑞滨(NVB)为新一代长春碱类抗肿瘤药,以长春瑞滨为主的联合化疗方案广泛应用于非小细胞肺癌、转移性乳腺癌、恶性淋巴瘤、卵巢癌等治疗.该药为强刺激、发疱性药物,对血管有很强的刺激性和损伤性,易渗人皮下间隙,引致局部浓度过高、pH值改变、静脉及毛细血管痉挛,组织缺血缺氧,导致静脉炎发生.     

Rh-Apo2L联合长春瑞滨杀伤人肺腺癌A549细胞的作用研究

[目的]探讨重组人凋亡素2配体(Rh-Apo2L)联合长春瑞滨(NVB)对人肺腺癌A549细胞的杀伤作用及机制。[方法]人肺腺癌A549细胞分成4组:对照组、Rh-Apo2L组、NVB组、联合组。用MTT法检测不同浓度Rh-Apo2L和NVB对A549细胞的抑制率,绘制细胞生长曲线,流式细胞仪测定细胞周期分布,RT-PCR方法检测死亡受体DR5基因mRNA的表达。[结果]MTT结果显示不同浓度Rh-Apo2L和NVB对人肺癌A549细胞的体外抑制作用均呈剂量效应关系,Rh-Apo2L处理48h后的IC20为3.48μg/ml、IC50为197.92μg/ml,NVB处理48h后的IC20为0.29μg/ml、IC50为3.44μg/ml。联合组对A549细胞的抑制作用高于单药组(P=0.001)。流式细胞分析显示含NVB药物两组在24hG2/M期比率高于对照组。3个实验组在24h、48h、72h后DR5基因mRNA相对表达量均较对照组多(P<0.05),但各实验组间均无统计学差异(P>0.05)。[结论]Rh-Apo2L与NVB联合应用能协同增强对人肺腺癌细胞株A549的体外抗肿瘤活性,其协同作用机制可能与细胞周期阻滞在G2/M期有关,但与DR5基因mRNA的表达无相关性。     

透明质酸修饰的pH响应型阿霉素纳米递送系统靶向多发性骨髓瘤的体外研究

目的:构建一种特异性靶向多发性骨髓瘤的pH响应型纳米递送系统,实现对多发性骨髓瘤细胞靶向释放化疗药物阿霉素。方法:采用酸敏感的DSPE-PEOz和阳离子类脂DOTAP包封模型药物阿霉素（doxorubicin,DOX）,得到阿霉素纳米递送系统（DOX-NDS）,并经HA-PEG2000-DSPE（HA）靶向长循环修饰,最终制得阿霉素靶向纳米递送系统（DOX-HA-NDS）;纳米粒度电位仪分析DOX-HA-NDS的粒径和Zeta电位;多功能酶标仪检测DOX-HA-NDS的包封率（encapsulation efficiency,EE）和载药量（drug loading,DL）;透析法研究DOX-HA-NDS在pH 5.0和pH 7.4条件下的体外释药行为;流式细胞仪检测其细胞摄入;CCK-8法评价空白靶向纳米递送系统（Blank-HA-NDS）的毒性和DOX-HA-NDS的细胞增殖抑制作用。结果:构建的DOX-HA-NDS粒径为（193.1±5.0）nm,Zeta电位为（-41.1±2.0）mV,离心法和透析法测得包封率分别高达93%和90%,载药量为32.76%和32%,4 ℃存储能稳定6个月以上。在pH 7.4条件下,DOX-HA-NDS的释药缓慢,且6 h累积释放率仅为30%,而在pH 5.0条件下,DOX-HA-NDS的释药明显加快,6 h累积释放率高达97%,表明构建的阿霉素靶向纳米递送系统具有pH响应控释性能。细胞摄入实验结果表明,经HA修饰后的阿霉素纳米递送系统可以更有效靶向肿瘤细胞;体外抗肿瘤活性结果表明,Blank-HA-NDS基本无细胞毒性,DOX-HA-NDS对人多发性骨髓瘤细胞（ARH-77）的增殖抑制作用最强。结论:构建的阿霉素靶向纳米递送系统不仅包封率高,而且兼具主动靶向和pH响应控释性能,提高对DOX的输送效率,从而增强了其对ARH-77的细胞增殖抑制作用。     

Comparison of free and liposome encapsulated doxorubicin tumor drug uptake and antitumor efficacy in the SC115 murine mammary tumor

Tumor drug uptake and antitumor efficacy of free and liposomal doxorubicin (DOX) were determined in the SC115 Shionogi mouse mammary tumor. Liposomal DOX systems were prepared by pH gradient-driven drug encapsulation in 170 nm egg phosphatidylcholine/cholesterol (55:45, mol ratio) vesicles. Intravenous injection of free DOX at 6.5 mg/kg, the maximum tolerated dose for free drug in the multiple dose therapy regimen, resulted in tumor-associated drug levels of 2.0 micrograms/g tissue at 1 h which remained constant over 24 h. Liposomal DOX injected at 6.5 mg/kg led to an accumulation of drug in the tumor from 2.6 micrograms/g tissue to 5.5 micrograms/g tissue between 1 h and 24 h, respectively. Increasing the dose of liposomal DOX to 13.0 mg/kg increased tumor drug uptake levels to 5.7 micrograms/g and 10.2 micrograms/g tissue at 1 h and 24 h, respectively. Administration of free or liposome encapsulated DOX every 7 days for 3 weeks resulted in a dose-dependent decrease in tumor growth rate. However, liposomal DOX injected at 6.5 mg/kg exhibited enhanced tumor growth inhibition compared to an equivalent dose of free drug. Further, the ability to administer increased doses of the less toxic liposomal DOX not only resulted in a greater inhibition of tumor growth but also significantly reduced tumor weight. Tumors weighing as much as 5 g were diminished to less than 0.5 g upon treatment with liposomal DOX at a dose of 13 mg/kg. In addition, groups receiving the highest liposomal DOX dose exhibited 25% complete tumor regression which persisted over the 50-day study period. These results demonstrate the ability of appropriately designed liposomal DOX systems to significantly enhance the delivery and retention of drug at solid tumor sites, resulting in increased therapeutic activity.     

Phase I study of vinorelbine and irinotecan in previously untreated patients with advanced non-small cell lung cancer

Introduction Vinorelbine alone and irinotecan alone have been shown to have efficacy against non-small cell lung cancer (NSCLC); each drug has different mechanisms of action. A phase I study using a combination of vinorelbine and irinotecan as first-line treatment for advanced NSCLC was done to determine the maximum tolerated dose (MTD) and the dose-limiting toxicity (DLT). Methods Previously untreated patients (≤75 years old) with Stage IIIB or IV NSCLC were enrolled. Based on a 4-week cycle, vinorelbine was given on days 1 and 8, and irinotecan was given on days 1, 8, and 15 intravenously. To prevent an injection site reaction to vinorelbine, the site was treated with topical clobetasol ointment, and the patients were given intravenous dexamethasone prior to vinorelbine treatment. DLT was defined as grade 4 neutropenia lasting ≥4 days or febrile neutropenia, grade 4 thrombocytopenia, ≥grade 3 non-hematological toxicities, or the need to cancel drug administration on both days 8 and 15. Results A total of 23 patients were enrolled. DLT was observed in 1 of 6 patients at level 3 (20 mg/m² vinorelbine, 50 mg/m² irinotecan), in 2 of 3 at level 4 (25 mg/m², 50 mg/m²), and in 2 of 5 at modified level 4 (20, 60 mg/m²). Level 4 and modified level 4 were considered to be the MTD; dose level 3 was therefore recommended. DLTs included liver dysfunction, pneumonitis, colitis, and arrhythmia. Injection site reactions were mild. Hematological and non-hematological toxicities were mild and easily controlled. Conclusion Use of 20 mg/m² vinorelbine on days 1 and 8 followed by 50 mg/m² irinotecan on days 1, 8, and 15 every 4 weeks warrants a phase II study.     

Efficacy of transferrin-conjugated paclitaxel-loaded nanoparticles in a murine model of prostate cancer

Chemotherapy remains the preferred choice of treatment for prostate cancer but modest drug response and significant toxicity by conventional methods of administration limit their efficacy. In our study, we determined the efficacy of paclitaxel (Tx)-loaded biodegradable nanoparticles (NPs) on tumor inhibition. We hypothesized that NPs following conjugation to transferrin (Tf) ligand (NPs-Tf) would enhance the therapeutic efficacy of the encapsulated drug. The antiproliferative activity of NPs was determined in human prostate cancer cell line (PC3) and their effect on tumor inhibition in a murine model of prostate cancer. NPs (approximately 220 nm in diameter, 5.4% w/w drug loading) under in vitro conditions exhibited sustained release of the encapsulated drug (60% release in 60 days). The IC50 (concentration of drug for 50% inhibition of cell growth) of the drug with Tf-conjugated NPs (Tx-NPs-Tf) was about 5-fold lower than that with unconjugated NPs (Tx-NPs) or drug in solution. Animals that received a single-dose intratumoral injection of Tx-NPs-Tf (Tx dose= 4 mg/kg) demonstrated complete tumor regression and greater survival rate than those that received either Tx-NPs or Tx-Cremophor EL formulation. In conclusion, sustained release NPs demonstrated greater antitumor activity following their conjugation to Tf ligand.     

奈达铂联合诺维本治疗晚期食管癌的临床疗效观察

目的:观察奈达铂联合诺维本治疗晚期食管癌的疗效和不良反应.方法:晚期食管鳞癌34例,奈达铂80-100mg/m2加入NS 500ml静脉滴注2h以上,第1天;诺维本25mg/m2加入NS 40ml中静脉推注,第1、8天,21天为1周期,连用2个周期后评价疗效.结果:全组 34例均可评价疗效及不良反应,总有效率 58.8%.其中初治21例中,PR 14例,有效率66.7%;复治13例中,PR 6例,有效率46.2%.主要不良反应为骨髓抑制,7例(20.6%)患者出现Ⅲ-Ⅳ度白细胞下降,3例(8.8%)患者出现Ⅲ-Ⅳ度血小板下降.消化道反应轻,未发现肝肾功能损害.结论:奈达铂联合诺维本治疗晚期食管癌疗效肯定,不良反应小,值得临床推广使用.     

Development of a highly active nanoliposomal irinotecan using a novel intraliposomal stabilization strategy

Liposome formulations of camptothecins have been actively pursued because of the potential for significant pharmacologic advantages from successful drug delivery of this important class of anticancer drugs. We describe nanoliposomal CPT-11, a novel nanoparticle/liposome construct containing CPT-11 (irinotecan) with unprecedented drug loading efficiency and in vivo drug retention. Using a modified gradient loading method featuring a sterically hindered amine with highly charged, multivalent anionic trapping agents, either polymeric (polyphosphate) or nonpolymeric (sucrose octasulfate), liposomes were capable of entrapping CPT-11 at extremely high drug-to-lipid ratios (>800 g CPT-11/mol phospholipid) and retaining encapsulated drug in vivo with a half-life of drug release in the circulation of 56.8 hours. CPT-11 was also protected from hydrolysis to the inactive carboxylate form and from metabolic conversion to SN-38 while circulating. The maximum tolerated dose in normal mice was determined to be 80 mg/kg for free CPT-11 and >320 mg/kg for nanoliposomal CPT-11. Nanoliposomal CPT-11 showed markedly superior efficacy when compared with free CPT-11 in human breast (BT474) and colon (HT29) cancer xenograft models. This study shows that intraliposomal stabilization of CPT-11 using a polymeric or highly charged, nonpolymeric polyanionic trapping agent results in a markedly active antitumor agent with low toxicity.     

Therapeutic efficacy of a polymeric micellar doxorubicin infused by convection-enhanced delivery against intracranial 9L brain tumor models

Convection-enhanced delivery (CED) with various drug carrier systems has recently emerged as a novel chemotherapeutic method to overcome the problems of current chemotherapies against brain tumors. Polymeric micelle systems have exhibited dramatically higher in vivo antitumor activity in systemic administration. This study investigated the effectiveness of CED with polymeric micellar doxorubicin (DOX) in a 9L syngeneic rat model. Distribution, toxicity, and efficacy of free, liposomal, and micellar DOX infused by CED were evaluated. Micellar DOX achieved much wider distribution in brain tumor tissue and surrounding normal brain tissue than free DOX. Tissue toxicity increased at higher doses, but rats treated with micellar DOX showed no abnormal neurological symptoms at any dose tested (0.1-1.0 mg/ml). Micellar DOX infused by CED resulted in prolonged median survival (36 days) compared with free DOX (19.6 days; p = 0.0173) and liposomal DOX (16.6 days; p = 0.0007) at the same dose (0.2 mg/ml). This study indicates the potential of CED with the polymeric micelle drug carrier system for the treatment of brain tumors.     

Analysis of the effect of liposome encapsulation on the vesicant properties,acute and cardiac toxicities,and antitumor efficacy of doxorubicin

Summary Numerous studies have demonstrated that liposomal encapsulation decreases the life-threatening chronic and acute toxicities of doxorubicin in the face of unaltered or improved antitumor activity. Minimal attention has been paid to the encapsulation effect on the lesser toxicities of the drug, specifically the vesicant properties. In this report we assess the effect of the encapsulation of doxorubicin in an egg-yolk phosphatidylcholine (EPC) cholesterol liposome on the drug's topical toxicity. In addition, to ensure acceptable activity and reduction in toxicity comparable with those of previously assessed formulations, the cardiac and acute toxicities and antitumor activity of the liposomal doxorubicin complex were also investigated. Antitumor efficacy was assessed using the metastatic murine P815 mastocytoma model. Equivalent doses of free and encapsulated doxorubicin possessed the same antitumor activity in the prolongation of animal survival in 14-day survival studies conducted to assess the effect of liposomal encapsulation on the acute toxicity of this drug. The LD₅₀ of liposomal doxorubicin was found to be 40 mg/kg, 53% higher than that of free doxorubicin (26 mg/kg). Histologic examination of cardiac sections taken from DBA/2J mice 7 days after a single i.v. injection of free or liposomal doxorubicin (25 mg/kg) revealed that the liposomal preparation was much less cardiotoxic. In animals receiving the free drug, edema, monocytic infiltration, and cell necrosis were evident. In contrast, those receiving the liposomal preparation demonstrated slight cellular edema but showed no evidence of cellular necrosis. To assess vesicant properties, DBA/2J mice were given a single s.c. injection (0.2 ml) of free or loposomal doxorubicin (2 mg/ml). Those receiving the free drug immediately developed erythema and edema at the injection site, which progressed to ulceration. Those receiving the liposomal complex developed slight erythema and edema but did not ulcerate at any time. All signs of irritation in this group had subsided 3 weeks postinjection. In summary, the liposomal complex used eliminated the vesicant properties of doxorubicin as well as significantly decreasing its cardiac and acute toxicities in the face of unaltered antitumor activity.