CD3AK细胞的体外诱导及免疫生物学特征的实验研究

目的：研究ＣＤ３ＡＫ细胞的体外诱导方法及其免疫生物学特征。方法：采用抗单克隆抗体（ａｎｔｉ－ＣＤ３ＭｃＡｂ）和基因重组人白细胞介素２（ｒＩＬ－２）、植物血凝素（ＰＨＡ０共同诱导人外周血单核细胞（ＰＢＭＣｓ）制备ＣＤ３ＡＫ细胞,应用ＡＰＡＡＰ法、吉姆萨染色法分析ＣＤ３ＡＫ细胞的表型、核型等免疫生物学特征。结果：微量的ａｎ－ｔｉ－ＣＤ３ＭｃＡｂ（终浓度３０～５０００ｎｇ／ｍｌ）辅以少量的ｒＩＬ－２（１     

细胞色素P4501A1,2D6和谷胱甘肽硫转移酶基因多态性与肺癌易感性的研究

目的探讨细胞色素Ｐ４５０１Ａ１（ＣＹＰ１Ａ１），２Ｄ６（ＣＹＰ２Ｄ６）和谷胱甘肽硫转移酶（ＧＳＴＭ１）基因多态性与肺癌易感性的关系。方法用病例对照研究方法及ＰＣＲ┐ＲＦＬＰ等技术检测原发性肺癌和住院对照各５９例，分析ＣＹＰ１Ａ１基因ＭｓｐＩＣ型、ＣＹＰ２Ｄ６Ｃｈ型（Ｔ／Ｔ型）和ＧＳＴＭ１缺陷型〔ＧＳＴＭ１（－）〕三种纯合突变型频率分布及其交互作用。结果突变型在病例和对照组的频率分别为（ＣＹＰ１Ａ１ＭｓｐＩＣ型２５．４％、１５．３％（Ｐ＝０．１７），ＣＹＰ２Ｄ６ＣｈＴ／Ｔ型３５．６％，４７．５％（Ｐ＝０．２６），ＧＳＴＭ１（－）型５７．６％、４９．２％（Ｐ＝０．４６），无显著性差异。协同分析发现在男性中，１１．６％（５／４３）肺癌兼有ＭｓｐＩＣ型和ＧＳＴＭ１（－）型，对照组无１例（０／４３），Ｐ＝０．０３。结论结果提示在男性中ＣＹＰ１Ａ１ＭｓｐＩＣ型和ＧＳＴＭ１（－）型可能协同增加患肺癌的危险性。     

GENOTOXIC EFFECT OF CDOP ON GERM CELL OF MALEMOUSE

无机抗癌剂氯氨顺铂（ＣＤＤＰ）可诱发小鼠初级精母细胞染色体畸变（ＰＳＣＡ）和精子畸形（ＳＡＭ），其诱发率分别在２．５－１５．０ｍｇ／ｋｇ和５．０－２０．０ｍｇ／ｋｇ体重剂量范围内具有剂量依赖关系。ＣＤＤＰ锈发ＰＳＣＡ和ＳＡＭ的最低检出剂量（ＭＤＤ）分别５．３８和５．４０ｍｇ／ｋｇ体重，两者在统计学上无明显差异，但ｌｍＭ／ｋｇ体重的ＣＤＤＰ诱发的ＰＳＣＡ和ＳＡＭ的细胞个数差异很大，前者为３．９６×１０￣１７个，后者为４１５．４２个，这可能是由于ＳＡＭ试验从给药至取样的时间间隔较ＰＳＣＡ试验者长，大量的受损生殖细胞丧失的关系。     

5-氟尿嘧啶、长春新碱与去甲二氢愈创木酸配伍对人恶性胶质瘤细胞株的作用

 目的 观察体外应用去甲二氢愈创木酸（ＮＤＧＡ）、予氟尿嘧啶（５－Ｆｕ）、长春新碱（ＶＣＲ）三种药物单用或合用对人恶性胶质瘤细胞系 ＳＨＧ－４４细胞的作用，并探讨其作用的可能机制。方法用 ＭＴＴ法检测药物作用效应，用免疫组化染色法检测细胞 ｃｙｃｌｉｎ Ｄ１基因的蛋白表达情况。结果 （１）三种药物单用及两药合用时随着药物浓度的增加其抗肿瘤效应也增加，且两药合用时药物的效应增强；（２）先给ＮＤＧＡ ２４ｈ后再给 ５－Ｆｕ或 ＶＣＲ，与同时给药对该细胞的抗肿瘤效应无显著差异（Ｐ＞０．０５），而先给 ５－Ｆｕ或 ＶＣＲ ２４ｈ后再给 ＮＤＧＡ，与同时给药对细胞的抗肿瘤效应有显著差异（Ｐ＜０．０５）；（３）免疫组化染色结果表明，与对照组相比，ＮＤＧＡ处理后该细胞 ｃｙｃｌｉｎ Ｄ１基因的蛋白表达明显降低。结论 ＮＤＧＡ与 ５Ｆｕ或 ＶＣＲ间有协同作用，且这种作用可能与 ＮＤＧＡ降低细胞 ｃｙｃｌｉｎ Ｄ１基因的蛋白表达有关。     

半边旗抗肿瘤有效成分6F对HL-60细胞周期的影响及体外增效作用

目的：研究半边旗抗肿瘤有效成分６Ｆ对ＨＬ－６０细胞周期的影响及对常用抗肿瘤药的体外增效作用。方法：应用流式细胞光度术（ＦＣＭ）测定细胞周期，应用噻唑蓝（ＭＴＴ）法测定药物对细胞的抑制率。结果：不同浓度６Ｆ作用６小时即可使ＨＬ－６０细胞Ｓ期及Ｇ２／Ｍ期比例升高，Ｇ１期比例下降，并呈一定的剂量效应关系，当作用到２４小时后，Ｓ期比例进一步升高，但Ｇ２／Ｍ期比例稍有回落。低浓度６Ｆ分别与２－氯代脱氧腺苷（２－ＣＬｄＡｄｏ），顺铂（ＣＤＤＰ），长春新碱（ＶＣＲ），氟尿嘧啶（５ＦＵ）合用可增强它们对ＨＬ－６０细胞的杀伤作用，ｑ值大于０．８５，与各药有相加或协同作用，６Ｆ对２－ＣｌｄＡｄｏ，ＣＤＤＰ，ＶＣＲ，５ＦＵ的增效倍数分别为１．５８，１．５３，１．５５，１．３８。结论：６Ｆ可明显阻断ＨＬ－６０细胞在Ｓ期及Ｇ２／Ｍ期；６Ｆ可增强上述药物对ＨＬ－６０细胞的杀伤作用。已知，２－ＣｌｄＡｄｏ阻断细胞在Ｓ期，ＣＤＤＰ和ＶＣＲ阻断Ｇ２／Ｍ期，５ＦＵ阻断Ｇ１期。鉴于所试药物对细胞周期的影响不同，提示６Ｆ的体外增效作用可能与此有关。     

耐顺铂人肺腺癌细胞系A549DDP的建立及耐药机制

采用递增顺铂（ＤＤＰ）浓度的方法，体外连续培养建成一株耐ＤＤＰ的人肺腺癌细胞系Ａ（５４９）ＤＤＰ，耐ＤＤＰ为亲代Ａ（５４９）的２４．４倍。在无ＤＤＰ的培养基中培养５月余，其耐药性仍稳定。Ａ（５４９）ＤＤＰ细胞内谷胱甘肽（ＧＳＨ）水平显著高于亲代细胞（Ｐ＜０．０１）。ＢＳＯ耗竭细胞内ＧＳＨ后，Ａ（５４９）ＤＤＰ细胞对ＤＤＰ敏感性增加５倍，ＢＳＯ对亲代Ａ（５４９）细胞无增敏作用，Ａ（５４９）ＤＤＰ细胞ＧＳＴ酶同功酶ＧＳＴ—π含量较Ａ（５４９）高１．６倍，却无ＧＳＴ基因扩增，表明ＧＳＨ／ＧＳＴ解毒系统参与Ａ（５４９）ＤＤＰ耐药性的产生。实验结果亦示Ａ（５４９）ＤＤＰ与卡铂及氨甲喋呤间存在交叉耐药，而与易产生ＭＤＲ或ａｔＭＤＲ之ＡＤＭ、ＶＣＲ、ＶＰ－１６、ＶＭ－２６无交叉耐药，Ａ（５４９）细胞无Ｐ－糖蛋白（Ｐ－ｇｐ）表达，Ｓｏｕｔｈｅｒｎｂｌｏｔ研究Ａ（５４９）ＤＤＰ无ｍｄｒｌＴｏｐｏⅡ基因扩增，提示Ａ（４５９）ＤＤＰ细胞系与ＭＤＲ及ａｔ－ＭＤＲ无交叉耐药。     

重组人肿瘤坏死因子抗卵巢癌的体内外实验研究

以ＭＴＴ法测试发现重组人肿瘤坏死因子（ｒＨＴＮＦ－α）对体外培养的人卵巢癌细胞系ＯＶＣＡＲ３和ＣＡＯＶ３有较强的细胞毒效应。同时还测试了五种化疗药物５－Ｆｕ、ＣＴＸ、ＶＣＲ、ＤＤＰ、ＫＳＭ对这两种细胞的细胞毒性。在化疗药物浓度参照临床用药剂量制定的条件下，五种化疗药物对这两种细胞的细胞毒性均明显低于ｒＨＴＮＦ－α。因此，ｒＨＴＮＦ－α抗卵巢癌的潜力值得重视。形态学研究表明，ｒＨＴＮＦ－α首先引起卵巢癌细胞的线粒体和内质网的破坏，继之才出现细胞膜损伤和细胞裂解。实验还证实，ｒＨＴＮＦ－α与ＤＤＰ或ＫＳＭ联合均有一定的协同抗癌效应。在体外实验的基础上，作者对人卵巢癌细胞ＯＶＣＡＲ３的裸鼠移植瘤模型进行了实验性治疗。结果显示，ｒＨＴＮＦ－α单独应用有显著的抗移植瘤作用，与ＫＳＭ合用有较强的协同效应。     

自体可溶性肿瘤抗原协同IL-2诱导外周血PBMC增殖及杀瘤

探讨自体可溶性肿瘤抗原（ＴＳＡ）与ＩＬ┐２共同诱导ＰＢＭＣ增殖及杀瘤作用。方法分三组用ＡＰＡＡＰ法观察细胞表型，ＭＴＴ法测定细胞增殖及杀瘤活性（靶细胞为ＭＮＫ４５系人胃低分化腺癌细胞）。结果培养第７天ＴＳＡ（１０μｇ／ｍｌ）协同ＩＬ┐２诱导ＰＢＭＣ增殖显著高于单独ＩＬ┐２诱导（Ｐ＜０．０１）。且ＣＤ＋８及ＣＤ＋２５表达率增高，差异有显著性（Ｐ＜０．０１０，当效／靶比为８０∶１时ＩＬ┐２诱导的ＰＢＭＣ杀瘤活性为４８．６％，而１０μｇ／ｍｌＴＳＡ协同诱导ＰＢＭＣ杀瘤活性为８６．３％。结论为研究ＴＡＫ细胞提供了实验依据，提示该类细胞具有临床过继免疫治疗应用的潜能     

5—氟脲嘧啶对耐顺铂人肺腺癌细胞A549^DDP的程序依赖性逆转作用

用耐顺铂（ＣＤＤＰ）人肺腺癌细胞系Ａ５４９ＤＤＰ作模型，研究了５－ＦＵ对ＣＤＤＰ耐药的程序依赖性逆转作用。５－氟脲嘧啶（５－ＦＵ）预处理Ａ５４９ＤＤＰ２４ｈ后立即给予ＣＤＤＰ，ＣＤＤＰ细胞毒性增加３．９倍；５－ＦＵ预处理Ａ５４９ＤＤＰ后间隔２４或４８ｈ再给予ＣＤＤＰ，其细胞毒性分别增加２０倍和２５０倍，甚至较亲代细胞Ａ５４９更为敏感；如先给ＣＤＤＰ后给５－ＦＵ则细胞毒性仅增加１．８倍。５－ＦＵ对亲代细胞亦有类似效应但细胞毒性增加程度明显低于耐药细胞。５－ＦＵ预处理后间隔２４，４８ｈ，细胞内ＧＳＨ含量逐渐降低，与细胞毒性逐渐增加相一致。如用ＢＳＯ耗竭Ａ５４９ＤＤＰ细胞内ＧＳＨ，ＣＤＤＰ细胞毒性增加６．４倍，仅能部分逆转ＣＤＤＰ耐药。５－ＦＵ明显抑制ＭＲＰ的表达，但对ＧＳＴπ的表达无影响。在５－ＦＵ预处理后的无药间隔时间内，给予无毒浓度的三苯氧胺则有明显的协同效应。结论：程序性给予５－ＦＵ通过降低细胞内ＧＳＨ含量和抑制ＭＲＰ的表达能完全逆转ＣＤＤＰ耐药     

CD3AK细胞和LAK细胞治疗晚期恶性肿瘤的临床和实验研究

将５１例晚期恶性肿瘤患者（男性２３例，女性２８例）分成两组，其中一组（３１例）以ＣＤ３ＭｃＡｂ（ＣＤ３单克隆抗体）和小剂量ＩＬ－２（５００ｕ／ｍｌ）共同诱导的ＣＤ３ＡＫ细胞治疗，另一组（２０例）输注大剂量ＩＬ－２（１０００ｕ／ｍｌ）诱导的常规ＬＡＫ细胞治疗，以探讨降低ＩＬ－２用量、提高杀伤细胞细胞毒活性的可能性。结果显示ＣＤ３ＡＫ组患者生活质量改善、症状缓解均优于ＬＡＫ组。ＣＤ３ＡＫ组ＰＲ＋ＭＲ率较ＬＡＫ组高２９．０％，Ｓ＋Ｐ率和死亡率分别较ＬＡＫ组低１２．４％和９．６％。同时比较了ＣＤ３ＡＫ细胞与ＬＡＫ细胞的体外增殖和细胞毒活性，结果表明ＣＤ３ＡＫ细胞增殖率高于ＬＡＫ细胞（Ｐ＜０．０１），靶细胞抑制率二者在０．０５水平无显著差异。提示ＣＤ３ＭｃＡｂ在刺激杀伤细胞活性，尤其在提高其增殖能力方面，具有显著的作用，ＣＤ３ＡＫ／ＩＬ－２能更有效地治疗晚期恶性肿瘤。     

A case of unresectable advanced gall bladder cancer successfully treated by hepatic arterial chemotherapy with reservoir (HACR) using CDDP and 5-FU

A 64-year-old female was admitted for treatment of a huge tumor (10 cm in diameter) in segment S4-S5 of the liver. The lymph node (2 cm in diameter) was located posterior to the pancreas head. The patient was diagnosed with an unresectable advanced gall bladder cancer with direct invasion of the liver bed and lymph node metastasis. At first, hepatic arterial infusion of CDDP, MMC and ADM through the hepatic artery was performed. Then, hepatic arterial chemotherapy with reservoir (HACR) using CDDP and 5-FU (CDDP 20 mg/body/day, 5-FU 750 mg/body/day) was started. As a result, the primary tumor and enlarged lymph node almost disappeared in two years. Mild bone marrow suppression, nausea and vomiting were encountered, but no severe side effects were noted. We conclude that this strategy is effective for unresectable advanced cancer of the gall bladder with lymph node metastasis.     

Two routes chemotherapy by CDDP and STS against liver tumor]

Nine (eight males, one female) patients with unresectable liver tumor (seven HCC and two metastasis) were treated by two-routes chemotherapy using cis-diamminedichloroplatinum (CDDP) and sodium thiosulfate (STS). In these patients, 50-100 mg/body of CDDP was administered through the proper hepatic artery or right hepatic artery by one shot infusion or the balloon-occluded arterial infusion (BOAI) at 10 mg/min, and during administration, intra-inferior vena cava injection of STS (4 g/body) was given. None of 9 patients suffered nausea and vomiting during the treatment, 3 of 9 patients suffered nausea and vomiting to a mild degree after the treatment, none of 9 patients showed significant side effects, such as bone marrow suppression and/or renal disfunction. In conclusion, this study demonstrated the protection effect of STS injected in inferior vena cava against the toxicity of CDDP were well indicated.     

Intra-arterial infusion chemotherapy with [Sar1, Ile8] angiotensin II in bladder cancer

Twenty patients with bladder cancer were treated with intra-arterial infusion chemotherapy using CDDP and ADM in combination with [Sar1, Ile8] angiotensin II. A catheter was introduced into internal iliac artery by Seldinger's technique, and 100 mg of CDDP, 50 mg of ADM and 1 mg of [Sar1, Ile8] angiotensin II were infused through the catheter for 40 minutes. CR was observed in 8 of 20 patients. PR in 11 and NC in 1. Therefore, the response rate (CR + PR) was 95% (19/20). Side effects were generally mild and consisted of leukopenia, nausea, vomiting, diarrhea, alopecia, skin pigmentation and headache. Catheter-related complications were not observed. This study demonstrated that intra-arterial infusion chemotherapy with CDDP and ADM in combination with [Sar1, Ile8] angiotensin II was extremely effective in treating patients with bladder cancer.     

Clinical study of the effect of cisplatin therapy in unresectable hepatocellular carcinoma

Twenty-one cases of unresectable hepatocellular carcinoma (HCC), including 15 cases receiving intravenous infusion of CDDP in addition to transcatheter arterial embolization (TAE), and 6 cases receiving intraarterial infusion of CDDP in combination with sodium thiosulfate rescue (STS rescue) were studied. In the 15 cases given intravenous infusion therapy with TAE, favorable effects were observed in 33.3% of patients, and the 50% survival period was 22.5 months. In the 6 cases given intraarterial infusion, favorable effects were obtained in 66.6% of patients, but the 50% survival period was 2 months. The side effects observed most frequently were nausea and vomiting. All the other side effects observed were not so severe. These results suggest that intravenous CDDP infusion in addition to TAE is favorable, producing a life-prolongation effect.     

人体肝癌细胞周期FCM测定与化疗药物敏感性关系的研究

应用细胞周期流式细胞仪(FCM)分析与~3H—TdR掺入法药敏测定技术,对8种化疗药物(MMC,MTX,5—Fu,ADM,CDDP,VP_(16),VCR,DTIC)在29例肝癌体外培养细胞的抑制作用与细胞周期比例的关系进行了对比研究。结果显示,化疗药物抑制率与细胞周期比例有关。以抑制率>60％作为有效标准,则对8种化疗药物有效的22例肝癌(75.9％)的细胞周期比例明显不同于抑制率<60％的肝癌。我们认为,FCM细胞周期测定具有简便、快速、客观、准确的特点,在临床个体化疗上具有实际应用价值。     

Apoptotic threshold of adriamycin and cisplatin in hepatocellular carcinoma

Chemotherapyisoneofmajorstrategiesagainsthepatocellularcarcinoma,andfailureofchemotherapyisusuallycausedbytoxicitiesofanticancerdrugsandmultidrugresistance.Recently,ithasbeenshownthatmanyanticancerdrugs,suchasADM,VCR,CDDP,MTX,etc.killtumorcellsbypromotingapoptosis.l]1Apoptoticthresholdisdefinedastheminimumdoseofanticancerdrugtocause,apoptosis.ThisstudywillinvestigatetheapoptoticthresholdofanticancerdrugsADMandCDDPinhepatocellularcarcinomainprimarycellculture.MATERIALSANDMETHODS…     

Combination chemotherapy of 5-fluorouracil (5-FU), adriamycin (ADM), cis-diamminedichloroplatinum (II) (CDDP) and mitomycin C (MMC) (FAP.MMC) in advanced gastric cancer

Twenty patients with advanced gastric cancer were treated with FAP.MMC (5-FU 350 mg/m2 i.v. on days 1-3, ADM 40 mg/m2 i.v. on day 1, CDDP 20 mg/m2 i.v. on days 1-3, MMC 6 mg/m2 i.v. on day 1), administering 5-FU, ADM and CDDP every 4 weeks and MMC every 8 weeks. Fourteen patients were evaluable for responses. Four (29%) partial responses and two minor responses were observed. The median duration of partial response was 3.8 months (range 2.5-7 months). The median overall survival time was 5 months (range 1.5-15 months). Leukopenia was relatively severe, with a median WBC nadir of 1,300/mm3. Nausea and vomiting were frequent but moderate. However, these toxicities were clinically manageable. FAP.MMC was thus considered effective for advanced gastric cancer.     

Non-surgical (medical) treatment of hepatocellular carcinoma (HCC)

Choice of treatment for HCC depends mainly on the size of tumor and patient's liver function because more than 80% of HCC patients are associated with liver cirrhosis. Percutaneous ethanol injection therapy (PEIT), transcatheter arterial embolization (TAE) and intraarterial infusion chemotherapy are, at present, commonly used treatments for HCC in Japan. PEIT is a safe and reliable treatment, in which absolute ethanol is injected to the tumor through a fine needle under US guide. PEIT is indicated for tumors of small size, which can not be removed surgically. The survival rate of PEIT for small liver cancer, less than 2 cm in diameter, is similar with the one of surgically removed cases. TAE is indicated for advanced HCC. Chemoembolization with Lipiodol is commonly used with good result. After TAE has been often performed, the survival rate of HCC patients was dramatically increased. In future, TAE combined with percutaneous transhepatic portal embolization or PEIT would be applied more often to obtain complete destruction of the lesion for advanced HCC. Intraarterial infusion chemotherapy is indicated for advanced HCC, in which TAE can not be performed. MMC, ADM and CDDP are commonly used anti-cancer drugs. Recently frequent infusion of these drugs has become possible by using implantable reservoir with good result. We have performed chemosensitivity test by SRCA for HCC specimens obtained by biopsy using a fine needle.     

Pilot combination phase II study of mitomycin C plus cisplatin for non-small cell lung cancer

Fifteen patients (six patients with adenocarcinoma, seven patients with squamous cell carcinoma, and two patients with large cell carcinoma) with advanced non-small cell lung cancer (NSCLC) were evaluable for mitomycin C (MMC; 8 mg/m2 day 1, 8, every 3-4 weeks) plus cisplatin (CDDP; 80 mg/m2 day 1, every 3-4 weeks). Ten patients had had prior chemotherapy. Among 15 evaluable patients, no patient achieved complete response, and two patients showed partial response. The response rate of MMC plus CDDP against NSCLC was 13.3%. Toxic effects included anorexia (80%), nausea and vomiting (67%), leukopenia (53%), anemia (47%), nephrotoxicity (47%), thrombopenia (27%), liver injury (27%), and fever (7%). These toxic effects were reversible and manageable. The combination of MMC and CDDP appears to be valuable regimen against advanced NSCLC.     

Antiemetic efficacy of granisetron: a randomized crossover study in patients receiving cisplatin-containing intraarterial chemotherapy

BACKGROUND: Cisplatin (CDDP) is one of the most active chemotherapeutic agents but is among the most emetogenic drugs. The emetic side-effects of CDDP-containing intraarterial chemotherapy have not been evaluated in a prospective randomized trial and the efficacy of serotonin antagonists in preventing the emesis associated with this method of CDDP administration has not been assessed. METHODS: CDDP 50 mg/m2 and methotrexate 30 mg/m2 were administered every 3 weeks through intraarterial catheters placed in the bilateral internal iliac arteries. Patients were classified into two groups: granisetron treatment group (group G) and no treatment group (group NG) with the first course of chemotherapy, crossing over with the second course. The patients in group G received granisetron 40 micrograms/kg by intravenous infusion. RESULTS: Although intraarterial CDDP administration produced less emesis than intravenous CDDP administration, at the same concentration, gastrointestinal toxicity is still the most unpleasant side-effect for patients. Granisetron administration significantly reduced nausea and vomiting during the acute emetic phase (an evaluation of treatment as very effective and effective was made in 89% in group G and 33% in group NG (P < 0.001). Complete control of emesis was achieved in 68 and 18% of patients in groups G and NG, respectively (P < 0.0001). CONCLUSION: A single prophylactic infusion of granisetron was effective in preventing the nausea and vomiting associated with intraarterial CDDP-containing therapy.