局部麻醉药对抗癌药物的体内外增效作用的研究

用体外培养的Eca-109细胞观察了阿霉素(ADM)和平阳霉素(PYM)单用或与局麻药普鲁卡因和利多卡因合用时的细胞毒作用,结果表明:局麻药在本身无细胞毒性浓度下与ADM或PYM联合应用时,可显著加强ADM和PYM的细胞毒作用,且随局麻药剂量增加增效作用加强,但各联合用药组之间的增效程度没有差异。普鲁卡因和利多卡因在本身无体内抑瘤活性的剂量下与顺铂合用,能显著地增强顺铂对实体型S_(180)肉瘤的抑瘤作用,并能延长荷S_(180)腹水瘤小鼠的生存期。     

CAC和CAP联合化疗方案对小鼠自发乳腺癌可移植模型抗瘤活性比较研究

 本实验应用小鼠可移植性自发乳腺癌瘤株，615近交系和津白Ⅱ近交系杂交F₁作为荷瘤动物。按10种治疗方案进行实验：对照DDP、CBP、ADM、CTX、CTX+CBP、CTX+DDP、CTX+ADM、CTX+ADM+CBP和CTX+ADM+DDP。动物体内试验结果表明：单药的抗瘤活性低，卡铂不如顺铂，三药联合作用明显提高，而CAC方案获得与CAP方案相似的抗瘤活性。故本实验提示可能用CAC方案代替CAP方案，使不能接受DDP的病人得到与用CAP方案相似效果。本实验为临床随机研究提供了科学依据，为乳腺癌治疗提供一个新的有效方案。     

人肺腺癌移植瘤多药耐药模型建立及生物学特性的研究

目的:建立耐药特性稳定的肺腺癌移植瘤裸小鼠动物模型,为进行耐药机制研究及逆转药物筛选提供实验模型和实验基础。方法:A549/DDP细胞接种于小鼠右侧腋下,观察肿瘤成瘤情况及生长特性;MTT法检测耐药倍数;RT-PCR和免疫组织化学染色检测小鼠移植瘤多药耐药1(MDR1)和多药耐药相关蛋白1(MRP1)的基因及蛋白表达。结果:移植瘤细胞和原代细胞对DDP的耐药倍数无差异。移植瘤成瘤率均为100%,于第6～9天成瘤(体积>100mm3)。与A549移植瘤相比,A549/DDP移植瘤成瘤潜伏期短(P=0.002),生长速度快。移植瘤中MDR1和MRP1 mRNA及P-糖蛋白(P-gp)和MRP1蛋白的相对表达量均升高,差异有统计学意义,P=0.000。结论:以A549/DDP细胞建立的肺癌多药耐药裸小鼠移植瘤模型,保持耐药活性和基因表型。     

环氧化酶-2抑制剂联合顺铂治疗肺癌的实验研究

目的 探讨环氧化酶-2(COX-2)抑制剂尼米舒利(NIM)与化疗药顺铂(DDP)联合应用治疗肺癌的效应及可能机制。方法 应用MTT试验及流式细胞术分别检测NIM与DDP联用对人肺腺癌A549细胞增殖和凋亡的影响，裸鼠移植瘤实验检测NIM与DDP联用对A549细胞成瘤性的影响。结果 在一定浓度范围内NIM、DDP均可抑制A549细胞的生长，其抑制作用呈量一效关系。NIM(25umol／L)与DDP合用可增强对A549细胞生长的抑制作用，DDP浓度≥1mg／L时二者有协同或相加作用。NIM及DDP可有效诱导A549细胞凋亡，联用后凋亡率显著增加。裸鼠移植瘤体内实验显示NIM与DDP均可抑制移植瘤生长，联用后抑瘤率显著增高。结论 NIM与DDP联用对A549细胞、肺癌移植瘤有显著的协同抗肿瘤效应，该作用可能是通讨增强对A549细胞增殖的抑制以及诱导其凋亡来实现的。     

rmhTNF α协同顺铂对小鼠Lewis肺癌移植瘤的抑制

目的:观察重组改构人肿瘤坏死因子(rmhTNF-α)协同顺铂(cisplatin ,DDP)对小鼠Lewis肺癌移植瘤的抑制作用,并探讨其可能的作用机制.方法:将Lewis肺癌细胞接种于C57BL/6小鼠右腋皮下,随机分为4组:生理盐水组(对照组)、DDP组(3 mg/kg)、rmhTNF-α组(150×104 U/kg)、rmhTNF-α(150×104 U/kg) DDP(3 mg/kg)组.于细胞接种的第7天,肿瘤直径约0.6 cm时,瘤内注射给药,连续3 d,停药1 d后处死小鼠,剥离并称取瘤重,计算抑瘤率.流式细胞术测定移植瘤细胞凋亡率及细胞周期, RT-PCR检测移植瘤组织Survivin的表达.结果:(1)rmhTNF-α DDP组治疗的抑瘤率(36.61%)明显高于DDP组(17.12%)或rmhTNF-α组(15.83%)单药治疗(P<0.05).两药联合使用的q=1.2,具有良好的协同作用.(2)联合用药治疗后移植瘤细胞的凋亡率[(28.2±1.8)%]显著高于DDP[(21.6±1.0)%]和rmhTNF-α[(19.3±2.0)%]单药治疗(P<0.05);前者的细胞周期明显阻滞于G2期.(3)3个治疗组移植瘤细胞Survivin 基因的表达受到明显抑制(P<0.01),联合用药组的抑制程度较单药组更明显(P<0.05).结论:rmhTNF-α与DDP联合应用具有协同抗Lewis肺癌移植瘤的作用,其机制可能与抑制Survivin基因表达、诱导肿瘤细胞凋亡有关.     

肿瘤坏死因子α和/或化疗药物联合应用对肝癌细胞系的协同细胞毒作用

本文采用体外细胞毒试验方法(MTT),检测了重组人肿瘤坏死因子α(rhTNF-α)和/或6种常用化疗药物包括阿霉素(ADM)、丝裂霉素(MMC)、顺铂(DDP)、卡铂(CBP)、足叶乙甙(VP-16)、异环磷酰胺(IFO).对肝癌细胞株SMMC-7721及BEL-7402的细胞毒效应.结果表明,rhTNF-α对两种肝癌细胞株的细胞毒作用有明显差异;两种肝癌细胞株对六种化疗药的敏感性不同,其中对ADM、DDP、MMC的敏感性较高.rhTNF-α与六种化疗药物联合应用的结果表明,rhTNF-α与ADM、DDP、VP-16有协同增强抗瘤效应,而rhTNF-α与MMC、CBP、IFO仅呈简单的叠加效应.体外细胞毒试验证明,rhTNF-α与某些化疗药物有协同作用,在临床肿瘤治疗中联合应用有可能提高疗效.     

左旋棉酚对小鼠宫颈癌移植瘤细胞凋亡的影响及其机制

目的探讨左旋棉酚［(-)-gossypol,LGP］对顺铂诱导小鼠宫颈癌移植瘤凋亡的影响及可能的作用机制。方法建立小鼠宫颈癌U14细胞皮下移植瘤模型,将60只小鼠按随机原则分为左旋棉酚组、顺铂(Cisplatin,DDP)组、左旋棉酚+顺铂(LGP+DDP)组及模型对照（Model Control）组4组,用药干预8天后采用电子天平称量瘤重并计算抑瘤率;采用TUNEL法检测各组瘤组织的凋亡情况;采用免疫组织化学法检测各组肿瘤组织中Bcl-2、Bax和P-gp蛋白的表达水平。结果各干预组小鼠移植瘤瘤重均低于Model组,其中LGP+DDP组瘤重最低,其次为DDP组(P＜0.01);各组移植瘤组织内AI均高于Model组（P＜0.05）,且LGP+DDP组最高（P＜0.01）;含LGP组移植瘤组织内P-gp、Bcl-2蛋白的表达降低（P＜0.05）, Bax蛋白的表达升高（P＜0.05）,Bax/Bcl-2比值升高,LGP+DDP组的上述作用明显优于单药组（P＜0.01）;相关性分析表明P-gp蛋白的表达量与Bcl-2的表达量呈正相关,与Bax的表达量呈负相关（P＜0.01）。结论 LGP能抑制小鼠宫颈癌移植瘤的生长,且与DDP联合后抑制作用更显著;其可能的机制:LGP可能是通过下调Bcl-2、上调Bax蛋白诱导小鼠宫颈癌细胞凋亡,降低P-gp蛋白的表达,增加肿瘤组织对DDP的化疗敏感度。     

中效原理在体外抗癌药物定量分析中的应用

 目的　在体外利用中效原理定量分析抗癌药物联合应用过程中的协同、相加或拮抗作用。方法　采用MTT法 ,利用中效原理判断联合用药 (阿糖胞苷 ,长春新碱 )对HL 6 0细胞的效应。结果　两种药物单用及联合应用时随药物剂量增加 ,其效应也随之增加。两药大剂量合用时为拮抗效应 ,小剂量合用时为协同效应。两药合用时给药次序不同不会影响合用效应 ,两药合用时药物浓度比例变化会影响合用效应。结论　两种药物合用时大剂量为拮抗 ,小剂量为协同 ,其效应大小与两种药物浓度比例有关 ,而与给药时间次序无关。     

以羟基喜树碱为基础的联合方案对实验性鼻咽癌抗瘤作用的研究

目的：探讨以拓扑异构酶Ⅰ抑制剂基对碱（10－hydroxycampothecin,HCPT)为基础的联合方案对实验性鼻咽癌（nasopharngeal carcinoma,NPC)的抗瘤作用。方法：以人NPC细胞株CNE2的体外和裸鼠移植瘤为模型，研究HCPT与顺铂（cisplatin,DDP)或（和）5－氟尿嘧啶（5－fluorouracil,5-FU)联合作用的抗瘤结果。细胞毒测定用MTT法，体内抗瘤模型为CNE2裸鼠模型。结果：HCPT、DDP和5－FU对CNE2的IC50分别为14．2、17．2及49．2μmol/L,HCPTgn DDP合用具有明显的协同抗NPC作用。HCPT与5－FU合用虽较单药抗瘤作用优，但未见明显的相加或协同作用。HCPT＋DDP＋5－FU三药合用药两药合用效果更优，且毒性可耐受。结论：HCPT＋DDP或HCPT＋DDP＋5－FU具有协同抗NPC肿瘤作用，可试用于临床。     

左旋棉酚对小鼠宫颈癌U14移植瘤的抑制作用和安全性的研究

目的：探讨左旋棉酚[（-）-gossypol,LG]对小鼠宫颈癌移植瘤的抑制作用,并观察其与顺铂（Cis-platin,DDP）联用的效果,同时观察左旋棉酚采用灌胃、腹腔注射两种途径给药对小鼠产生的不良反应。方法：建立小鼠宫颈癌U14细胞皮下移植瘤模型,按随机原则将90只小鼠分为左旋棉酚腹腔注射[（-）-gos-sypol interaperitonea Injection,LGIP]组、左旋棉酚腹腔注射＋顺铂（LGIP＋DDP）组、左旋棉酚口服[（-）-gossypol peros,LGPO]组、左旋棉酚口服＋顺铂（LGPO＋DDP）组、顺铂（DDP）组及模型对照组6组,连续用药干预8天,在用药第1、3、5、7天及停药2日后分别测量各组小鼠移植瘤体积并绘制生长曲线。停药后两日处死小鼠,剥离瘤体,称量瘤重并计算抑瘤率。密切观察用药期间各组小鼠一般情况的变化（包括体重、白细胞、脱毛）,同时记录小鼠死亡情况并分析死亡原因。结果：皮下注射宫颈癌U14细胞建立昆明小鼠皮下移植瘤模型,成瘤率达100%。各干预组小鼠移植瘤的体积及瘤重均低于模型对照组（P〈0.01）,LGPO＋DDP组小鼠移植瘤的体积及瘤重明显低于DDP组、LGPO组、LGIP组（P〈0.01）,DDP组小鼠移植瘤的体积及瘤重低于LGPO组、LGIP组（P〈0.01）,LGPO＋DDP组与LGIP＋DDP组、LGPO组与LGIP组之间的移植瘤的体积及瘤重无统计学差异（P〉0.05）;各干预组在用药开始后小鼠的体重均有不同程度的减轻（P〈0.01）,按减轻程度由大到小排序依次是LGIP＋DDP、LGPO＋DDP、DDP、LGIP、LGPO（P〈0.01）,其中,LGIP＋DDP与LGPO＋DDP、LGIP与DDP组小鼠体重减轻无统计学差异（P〉0.05）。DDP组、LGPO＋DDP组、LGIP＋DDP组小鼠白细胞水平明显低于模型对照组（P〈0.01）,DDP组小鼠白细胞水平低于LGPO＋DDP组（P〈0.05）,LGPO组、LGIP组与模型对照组的小鼠白细胞水平无统计学差异（P〉0.05）;LGIP组与LGIP＋DDP组小鼠的死亡率明显高于其它各干预组（P〈0.01）,且肠梗阻的发生率分别高达93.3%、53.3%,明显高于其它各干预组（P〈0.01）。结论：左旋棉酚对小鼠宫颈癌U14细胞株移植瘤具有明显抑制作用,与顺铂联合用药效果更显著。左旋棉酚经灌胃途径给药较腹腔注射更安全有效。     

Combination cytotoxic chemotherapy with cisplatin or doxorubicin and photodynamic therapy in murine tumors

This study was designed to evaluate the interaction of photodynamic therapy (PDT) and chemotherapy in an animal model. PDT is based on the interaction of hematoporphyrin derivative and red light of the appropriate wavelength (630 nm) and intensity. Two tumor models were utilized: C3H/Km mice bearing the RIF-1 tumor and BALB/c mice bearing the EMT-6 tumor. Tumor-bearing mice were treated with either cisplatin (DDP), doxorubicin (ADM), PDT, or a combination of drug and PDT. It was demonstrated that the RIF-1 tumor was sensitive to DDP and insensitive to both PDT and ADM. There was no additional antitumor effect when either drug was combined with PDT. The EMT-6 tumor was moderately sensitive to PDT and mildly sensitive to both DDP and ADM. Although the addition of DDP did not potentiate tumor destruction, the addition of ADM significantly enhanced the effect of PDT (P = .01). The enhanced activity of the combination of PDT and ADM appeared to be the result of increased activity of ADM alone, when illuminated with red (630 nm) light. This potentiation may be due to a photochemical process or may be secondary to the mild hyperthermia generated by illumination with the laser. This study demonstrates that PDT combined with cytotoxic chemotherapy is well tolerated in these animals and that certain combinations of PDT and chemotherapy may result in an enhanced tumoricidal effect.     

p73基因过量表达对人肺癌细胞A549凋亡及其化疗敏感性的作用

背景与目的：部分非小细胞肺癌（non-small cell lung cancer,NSCLC）表达野生型p53基因（wild-type p53,wt-p53）,因此在基因治疗中克服这些NSCLC对wt-p53的抵抗机制就非常重要。P53基因家族的新成员p73是p53的同源体,本研究旨在探讨对wt-p53基因治疗抵抗的人肺腺癌细胞A549在外源p73基因转染或联用化疗药后凋亡程度和对化疗药物敏感性的变化。方法：将真核表达重组质粒pcDNA3-HA-p53或pcDNA3-HA-p73α转染入A549细胞,G418筛选,Western blot检测P53或P73α的表达。MTT法分析转染细胞对顺铂和阿霉素的敏感性,用流式细胞术、TUNEL法和DNA片段法分析化疗药作用下转染细胞的凋亡变化,克隆形成实验观察细胞生物学性状的改变。结果：转染p53或p73α基因的A549细胞可以稳定高表达p53或p73α蛋白。转染p73α的A549细胞在原本没有明显抑制和杀伤作用的药物浓度（6．25μmol／L的顺铂或0．25μmol／L的阿霉素）作用下生长明显受到抑制。顺铂的IC50值从22．65μmol／L降至3．75μmol／L,阿霉素的IC50值从4．20μmol／L降至0．06μmol／L。p73能使A549细胞受顺铂和阿霉素诱导的细胞凋亡增加。而p53没有明显作用。流式细胞术显示p73α基因转染后,顺铂诱导的细胞凋亡率从10．6％升高到36．8％（P〈0．01）,阿霉素诱导的细胞凋亡率从13．0％升高到41．1％（P〈0．01）。克隆形成实验显示．p73α基因转染能明显降低顺铂和阿霉素作用后A549细胞的克隆形成数（P〈0．01）。对顺铂和阿霉素的化疗增效倍数分别为2．0和2．4倍。结论：外源性p73基因的导入增加了wt-p53型A549细胞对顺铂和阿霉素等化疗药的敏感性。该作用可能与p73基因能不依赖于p53基因诱导细胞凋亡有关。p73基因可用于治疗wt-p53不能发挥作用的恶性肿瘤。     

Effect of combined intrapleural administration of Lactobacillus casei (LC9018) and adriamycin on experimental malignant pleurisy in mice

The combined effect of Lactobacillus casei YIT9018 (LC9018) and adriamycin (ADR) on malignant pleurisy was investigated using an experimental model in BALB/c mice in which Meth A fibrosarcoma cells were intrapleurally implanted. The control mice died from dyspnea due to pleural effusion, before significant growth of tumor nodules could be achieved in the thoracic cavity. Intrapleural (ipl) administration of LC9018 (20-200 micrograms/head) on days 1 and 5 reduced the effusion volume and induced pleural adhesions in a dose-related manner. A statistically significant and reproducible prolongation of survival was observed at a dose of LC9018 200 micrograms/head: increase of lifespan (ILS) values of 15-39% were obtained. An ipl administration of ADR (2-4 mg/kg) on day 1 was also effective in prolonging survival without severe toxicity (ILS values of 100-122%). The combined use of ADR and LC9018 induced a high incidence of pleural adhesions, a delay in effusion accumulation, and an additive prolongation of lifespan (ILS values of 133-178%), compared with ADR monotherapy. In the combination therapy group, a marked and continuous ipl exudation of neutrophils, macrophages, and lymphocytes was observed with a significant decrease in pleural tumor cells. These findings suggest that ipl administration of LC9018 enhances the effect of ADR, probably through both host-mediated tumoricidal activity and sclerosing effects on the pleura.     

Effect of Combined Intrapleural Administration of Lactobacillus casei (LC9018) and Adriamycin on Experimental Malignant Pleurisy in Mice

The combined effect of Lactobacillus casei YIT9018 (LC9018) and adriamycin (ADR) on malignant pleurisy was investigated using an experimental model in BALB/c mice in which Meth A fibrosarcoma cells were intrapleurally implanted. The control mice died from dyspnea due to pleural effusion, before significant growth of tumor nodules could be achieved in the thoracic cavity. Intrapleural (ipl) administration of LC9018 (20–200 μg/head) on days 1 and 5 reduced the effusion volume and induced pleural adhesions in a dose-related manner. A statistically significant and reproducible prolongation of survival was observed at a dose of LC9018 200 μg/head: increase of lifespan (ILS) values of 15–39% were obtained. An ipl administration of ADR (2–4 mg/kg) on day 1 was also effective in prolonging survival without severe toxicity (ILS values of 100–122%). The combined use of ADR and LC9018 induced a high incidence of pleural adhesions, a delay in effusion accumulation, and an additive prolongation of lifespan (ILS values of 133–178%), compared with ADR monotherapy. In the combination therapy group, a marked and continuous ipl exudation of neutrophils, macrophages, and lymphocytes was observed with a significant decrease in pleural tumor cells. These findings suggest that ipl administration of LC9018 enhances the effect of ADR, probably through both host-mediated tumoricidal activity and sclerosing effects on the pleura.     

Antitumor activity of mitoxantrone against murine experimental tumors: Comparative analysis against various antitumor antibiotics

Summary 1,4-Dihydroxy-5,8-bis{{{{2-[(2-hydroxyethyl)amino]ethyl}amino}}-9,10-anthracenedione dihydrochloride (mitoxantrone) was tested for antitumor activity against experimental tumors in mice and the results were compared with those of seven antitumor antibiotics: adriamycin (ADM), daunomycin (DM), aclarubicin, mitomycin C (MMC), bleomycin, neocarzinostatin, and chromomycin A₃. The drugs were given IP or IV, in general on days 1,5, and 9 following tumor inoculation. Mitoxantrone given IP at the optimal dose (1.6 mg/kg/day; as a free base) produced a statistically significant number of 60-day survivors (curative effect) in mice with IP implanted L1210 leukemia. The curative effect was not observed with any of the other antibiotics. In the case of IV implanted L1210 leukemia, there was an increase in lifespan (ILS) by more than 100% in the mice following IV treatment with mitoxantrone or DM. In IP implanted P388 leukemia, the curative effect was elicited by IP treatment with mitoxantrone or MMC. In IP implanted B16 melanoma, both the curative effect and a more than 100% ILS in mice that did die were produced by IP treatment with mitoxantrone or ADM. In SC implanted Lewis lung carcinoma, mitoxantrone and ADM administered IV also showed effective antitumor activities and produced a 60% and a 45% ILS, respectively. In conclusion, mitoxantrone and ADM had a wider spectrum of antitumor activity against mouse tumors, including two leukemias and two solid tumors, than did the other drugs; however, mitoxantrone elicited higher antitumor effects than ADM on mouse leukemias, especially on L1210 leukemias. Moreover, mitoxantrone possessed much higher therapeutic indices than ADM against IP implanted P388 (optimal dose/ILS₄₀; >128 versus 15.2) and L1210 (optimal dose/ILS₂₅; 72.7 versus 4.8) leukemias. In addition, mitoxantrone showed moderate activity against DM-resistant L1210 leukemia.     

Mitomycin C combination therapy against murine tumor systems. Effectiveness with cyclophosphamide and methotrexate

Mitomycin C (MMC) has been evaluated in combination with several antitumor agents. Full dose response curves were established for all drugs and drug combinations. Synergy was shown with MMC plus either cyclophosphamide (CYC) or methotrexate (MTX). In testing MMC and CYC against P388 leukemia, the combined treatment yielded a 75% rate of long-term survivors at the optimal level, compared to no survivors at the optimal level of the best single agent, CYC, alone. There was no increased toxicity among the combination-treated animals. Large increases in lifespan were obtained against L1210 and B16. Maximally tolerated doses of the single agents could be combined without increased toxicity. The combination of MMC and MTX was synergistic against ip L1210 and P388 leukemias. The responses of mice bearing L1210 to treatment on days 1, 5, and 9 respectively, were 42% ILS for 3.0 mg/kg MMC; 96% ILS for 15 mg/kg MTX; 172% ILS with four out of ten survivors for 3.0 mg/kg MMC plus 15 mg/kg MTX. MMC and adriamycin (ADR) were found to be synergistic against B16 melanoma at one schedule but not against another schedule, or against colon carcinoma 26. No improvements over optimal nontoxic single agent therapy were seen for chlorambucil, 5-fluorouracil, dibromodulcitol, cis-diaminedichloroplatinum or 4-'(9-acridinylamino) methansulfon-M-anisidide. On the basis of these data, recommendations were made for clinical trials for MMC plus either CYC or MTX against lung, breast, and colon tumors.     

Schedule-dependent effectiveness of ADM and multiple cycles of DDP on a murine reticulum sarcoma

The therapeutic index of adriamycin (ADM) and cisplatin (DDP) combination versus repeated sequences of the more active drug (DDP) was investigated on a murine reticulum cell sarcoma of ovarian origin (M5) implanted i.m. in C57BL/6 mice. The antitumor efficacy of multiple cycles of DDP according to different regimens at a prefixed time (every 7 days) or at tumor regrowth was also evaluated. Our data demonstrate that the ADM-DDP combination did not improve the antitumoral efficacy of DDP as single agent, while repeated cycles of DDP led to a significant increase in the host life span. Differences in therapeutic effect were elicited by the two schedules: the regimen at a prefixed time showed a major effect on local tumor control, although the regimen at tumor regrowth was better tolerated.     

MK-2206联合顺铂对乳腺癌4 T1／DDP移植瘤耐药性的影响

目的：探讨蛋白激酶B（protein kinase B,Akt）抑制剂MK-2206联合顺铂对三阴性乳腺癌4T1顺铂耐药（4T1／cisplatin,4T1／DDP）移植瘤耐药性的影响。方法采用顺铂（cisplatin,DDP）低剂量诱导及体内外交叉致瘤相结合的方法建立三阴性乳腺癌4T1／DDP耐药小鼠移植瘤模型,给予DDP腹腔注射（0．3 g／L,1次／d）单独或联合MK-2206灌胃（12 g／L,1次／周）治疗。给药28 d后处死小鼠,小动物成像检测观察肿瘤生长情况;Western blot法检测肿瘤组织中磷酸化Akt（phosphorate-Akt,p-Akt）和总Akt（total-Akt,t-Akt）蛋白水平变化;免疫组织化学法分析耐药相关蛋白P-糖蛋白（P-glycoprotein,P-gp）、乳腺癌耐药蛋白（breast cancer resistance protein,BCRP）、基质金属蛋白酶7（matrix metalloproteinase 7,MMP7）表达差异。结果建立的三阴性乳腺癌耐药小鼠模型达中等耐药程度。建立非耐药小鼠与耐药小鼠肿瘤组织生长速度未见明显区别（P＞0．05）。分别给予这两种模型小鼠相同剂量（0．3 g／L）的DDP治疗后,耐药小鼠对DDP的敏感性低于非耐药小鼠（P＜0．01）。将MK-2206（12 g／L）与DDP （0．3 g／L）联合应用后小鼠肿瘤较单独应用DDP缩小,且可降低P-gp、BCRP、MMP7蛋白的表达（均P＜0．05）,并降低Akt蛋白磷酸化水平（P＜0．01）。结论 MK-2206联合顺铂能够逆转4T1／DDP移植瘤耐药性,且与抑制Akt蛋白磷酸化相关。     

靶向NAC1同源二聚体小分子抑制剂NIC19鉴定及其与化疗药物协同抗肿瘤作用研究

目的致癌转录因子的过度激活与肿瘤无限增殖、侵袭转移、耐药等恶性特征密切相关,靶向致癌转录因子为主要策略的肿瘤治疗日益受到关注。本课题基于NAC1蛋白二聚体结构进行了虚拟筛选,找到了另一个小分子先导化合物NIC19,并探究其药理作用及临床潜在意义。方法计算机辅助药物筛选发现靶向NAC1蛋白同源二聚体化合物NIC19;蛋白质印迹法检测NIC19的降解和表达;四甲基偶氮唑盐(methyl thiazolyl tetrazolium,MTT)法和克隆集落形成实验检测NIC19对抗肿瘤药物作用的影响;裸鼠皮下肿瘤模型进一步验证NIC19对抗肿瘤药物作用。结果NIC19的发现及计算机辅助NIC19与NAC1蛋白对接模型;NIC19可下调NAC1蛋白的表达和稳定性,呈现时间和剂量依赖性(时间:与0μmol/L表达量2.62±0.1相比,NAC1蛋白的表达量降至5μmol/L的1.85±0.13,10μmol/L的1.18±0.03,20μmol/L的0.65±0.03,差异有统计学意义,F=63.703,P<0.001;剂量:与0h表达量2.43±0.13相比,NAC1蛋白表达量降至12h的2.30±0.03,24h的2.02±0.04,48h的0.49±0.15,差异有统计学意义,H=10.385,P=0.016);NIC19可增加肿瘤细胞对顺铂(cisplatin,DDP)或多柔比星(doxorubicin,DOX)治疗的敏感性(与DOX组相比t=9.220,P=0.0002;与DDP组相比t=6.844,P=0.001)。同时,克隆集落形成实验表明,联合NIC19可增强在DOX和DDP治疗下抑制SKOV3克隆集落形成能力(FNIC19×DOX=16.472,P<0.001;FNIC19×DDP=15.412,P<0.001),并且DDP或DOX与NIC19联合使用时,SKOV3凋亡水平明增高,表现为凋亡蛋白cleaved-PARP水平升高(FNIC19×DDP=7.041,P=0.029;FNIC19×DOX=6.098,P=0.039)。在体研究进一步证实了NIC19可增强肿瘤细胞对DDP的敏感性,在皮下接种HeLa细胞的裸鼠中,NIC19和DDP联合使用可抑制移植瘤的生长,FNIC19×DDP=9.720,P=0.014。与NIC19或DDP单独使用组相比,联合用药组可增加瘤体凋亡的发生,表现为TUNEL染色阳性细胞数增加(H=10.421,P=0.015),增殖标志物Ki-67阳性率减少,H=9.425,P=0.024。结论通过计算机和高通量筛选得到具有预期药理活性的靶向该二聚体的小分子抑制剂NIC19,其可影响NAC1蛋白的同源二聚体形成,降低NAC1蛋白的表达和稳定性;进一步研究发现,NIC19可增敏DDP和DOX等传统化疗药物的抗肿瘤作用,同时在体研究进一步证实了这种现象,为进一步研究和开发该类小分子抑制剂提供理论基础。     

体外MTT法预测卵巢上皮癌化疗药敏性与体内疗效相关性的评估

目的：预测卵巢上皮癌体外化疗敏感性与体内疗效的关系。方法：运用MTT法检测7种临床常用化疗药物及6种经验组合方案对卵巢上皮癌的体外抑制率进行了检测并与体内疗效结果进行比较。结果：7种药物抑制率范围为6．25％～51．43％其中PTX为51．43％最高，TXT为20．29％，DDP为18．57％，TPT为11．63％，ADM为11．43％．GEM8．82％及VP166．25％敏感性最差；6种联合方案的抑制率范围在10．00％～35．00％．TPT＋PTX＋DDP（35．00％）及PTX＋DDP（30．00％）较好，其余4种TXT＋DDP（20．59％），TPT＋DDP（17．65％），GEM＋DDP（11．76％）及VP16＋ADM（10．00％）均较差。本组药物抑制率与试验前是否用过化疗药无关体内、外敏感及耐药的总符合率为72．86％（51／70），不符合率27．14％（19／70），统计学有显著性差异（P〈0．05）。结论：MTT法对卵巢上皮癌的个体化治疗计划有参考价值．能在一定程度上指导临床用药。