改良染料排斥法体外药物敏感试验预测急性白血病化疗的反应

使用改良染料排斥法测定１７例２０人次的白血病患者对常用的９种化疗药物体外的敏感性，其中急性白血病１４人，ＦＡＢ分型ＡＮＬＬ１０人，ＡＬＬ４人，ＭＤＳ－ＲＡＥＢＴ１人ＣＶＭＬ急变２人，体外药敏结果显示：体外药敏实验与体内疗效总符合率为５９．２％，阳性符合率６５．８２％，阴性符合率４８．９１％。初步表明法对指导临床白血病个体化治疗，选用有效的化疗药物有一定的应用价值。     

急性髓系白血病细胞四项耐药指标的评价

目的　为研究急性髓系白血病（ＡＭＬ）细胞耐药指标的敏感性和耐药方式。方法　ＭＴＴ药物敏感试验，白血病祖细胞（ＣＦＵＬ）体外生长类型，Ｂｃｌ２ 抗原表达和Ｂｃｌ２／Ｂａｘ 抗原比值，流式细胞仪测定细胞内柔红霉素（ＤＮＲ）荧光强度四项指标被综合评价。结果　６２ 例ＡＭＬ中，ＭＴＴ药敏试验阳性符合率为７３％ ，阴性符合率为７０％ ，三种临床常用药物中一种药物敏感预示缓解的符合率达７１％ 。５１例ＡＭＬ中，３１例完全缓解（ＣＲ）组中ＣＦＵＬ自主分泌生长２９例，不生长２例，２０ 例未缓解（ＮＲ）组中，自主分泌生长型１４ 例，不生长型６例，统计学有显著差异性（Ｐ＜ ０．０５）。３２ 例ＡＭＬ中，药敏组Ｂｃｌ２ 表达率为５９．５５％ ±１９．５６％ ，耐药组为７７．３６％ ±１１．９１％ （Ｐ＜ ０．０５），Ｂｃｌ２／Ｂａｘ 比值药敏组为７．５０±５．０４，耐药组为１４．３２±８．９９（Ｐ＜ ０．０５）。１５例临床耐药的ＡＭＬ，ＤＮＲ荧光直方图１２ 例呈现主峰左移，诊断为经典耐药，３ 例呈现主峰右移，诊断为再生耐药。结论　ＭＴＴ，ＣＦＵＬ检测可预示临床治疗是否耐药，Ｂｃｌ２，Ｂｃｌ２／Ｂａｘ 检测与患者预后有关，有ＤＮＲ直方     

急性白血病体外药敏试验MTT法的初步探讨

为了探讨肿瘤细胞体外药敏试验ＭＴＴ法的实验条件，观察体外药敏结果和体内疗效的相关性，作者采用微量液体培养和微量比色法测定了２３例急性白血病患者对常用的６种抗癌药物的体外敏感性，确定了６种化疗药物的体外作用浓度。结果显示，体外药敏试验的结果和体内疗效总符合率为８２．６％，阳性符合率为８８．９％，阴性符合率为７１．４％。提示ＭＴＴ法是一种简便、快速、客观的肿瘤细胞药敏试验方法，对指导急性白血病个体化治疗有较高的实用价值。     

卵巢恶性肿瘤体外药物敏感试验及其临床意义

目的：探讨卵巢恶性肿瘤体外药物敏感试验的临床意义。方法：采用四氮唑蓝显色反应药敏分析法（ＭＴＴ）检测６０例卵巢恶性肿瘤组织（Ⅰ期８例、Ⅱ期２例、Ⅲ期２２例、Ⅳ期１例、复发２７例）对化疗药物的敏感性。结果：体外试验单药对肿瘤的中位抑制率为：泰素６１．５％、表阿霉素５２．４％、卡铂４２．１％、平阳霉素３３．５％、长春新碱１０．７％。联合用药的中位抑制率为：ＶＢＰ（长春新碱＋平阳霉素＋卡铂）７９．９％、ＶＡＰ（长春新碱＋表阿霉素＋卡铂）７５．３％。ＶＢＰ、ＶＡＰ方案对上皮必卵巢产中位抑制率分别为７７．３％和７１．０％。初治术后残留灶≤２ｃｍ者中２６例可评价,体外试验与临床疗效的符合率为８４．６％。复发术后残留灶≤２ｃｍ者中１５例可评价,１０例使用体外敏感药物进行化疗,术后１年生存率８０．０％,５例患者体外试验不敏感     

健康人和骨肉瘤病人来源的LAK细胞体外抗瘤作用

采用５１Ｃｒ释放试验对健康人和骨肉瘤病人外周血单个核细胞（ＰＢＭ）在重组白细胞介素－２（ｒＩＬ－２）条件下，ＬＡＫ细胞的诱导形成及对４种传代瘤细胞的体外杀伤活性进行探讨。实验结果表明：２种来源ＬＡＫ细胞对Ｋ５６２（人慢性髓样红白血病细胞系）、ＳＭＭＣ７７２１（人肝癌细胞系）、ＬＡＸ（人肺腺癌细胞系）的杀伤活性均在５５％以上（按效靶比例５０：１），而对ＯＳ细胞（人骨肉瘤细胞系）的活性普遍低下，杀伤率低于３５％。结果证实：１）健康人和骨肉瘤病人的ＰＢＭ均能在ｒＩＬ－２条件下诱导形成具有广谱抗瘤活性的ＬＡＫ细胞群，二者的杀伤格局和效力相近。２）骨肉瘤细胞本身对ＬＡＫ细胞的杀伤作用存在强烈抗性。     

白血病细胞体外药敏试验的临床意义及其与m dr1表达关系的探讨

目的　探讨白血病细胞体外药敏试验的临床意义及从敏感性和耐药性两个不同侧面，研究两者内在联系。方法　采用ＭＴＴ法对２７例急性白血病患者的白血病细胞进行了Ａｒａ－Ｃ、ＤＮＲ、Ｈａｒ、ＭＩＴ、ＶＰ１６、ＶＭ２６、ＶＣＲ及ＨＣ等８ 种常用化疗药物的体外敏感试验以及用反转录－聚合酶链反应（ＲＴ－ＰＣＲ）方法检测了２２ 例患者多药耐药基因ｍ ｄｒ１ 的表达情况。结果　回顾性研究显示ＭＴＴ法体外药敏试验结果与临床疗效的总符合率为７７．３％ ，阳性符合率６１．５％ ，阴性符合率１００％ ，灵敏度１００％ ，特异度６４．３％ 。结论　ＭＴＴ法有一定的临床预测价值，是辅助临床用药选择的实验检测。ＭＤＲ相关药物ＤＮＲ、Ｈａｒ、ＭＩＴ、ＶＰ１６ 及ＶＭ２６ 体外药敏结果与ｍ ｄｒ１ 表达水平无相关性，提示可能存在其它的耐药机制。     

CD4抗原在急性髓系白血病细胞上的表达及其意义

目的：探讨ＣＤ４抗原在急性髓系白血病细胞上的表达及意义。方法：对８２例急性髓系白血病患者进行免疫表型、细胞遗传学分析。结果：ＣＥ４在ＡＭＬ患者中表达率为４０．２％，Ｍ５中阳性率最高９２．９％，Ｍ４次之为５５％，ＣＤ４＾＋ＡＭＬ高表达ＨＬＡ－ＤＲ，ＣＤ３８、ＣＤ３３、ＣＤ１５、ＣＤ１４、ＴＢ系列抗原阴性。ＣＤ４＾＋ＡＭＬ中可见１１ｑ２３、ｉｎｖ（１６）、ｔ（９；２２），未见特异性染色体异常。但伴１１ｑ２３和ｉｎｖ（１６）异常的ＡＭＬ频繁表达ＣＤ４，阳性率分别为８６．３％、６０．２％。ＣＤ４＾＋ＡＭＬ在年龄、性别、肝脾肿大、ＣＮＳ－Ｌ、ＤＩＣ、１疗程ＣＲ率等临床特征方面无明显不同。结论：ＣＤ４＾＋ＡＭＬ是一种起源较高的具有单核细胞特征的髓系白血病，ＣＤ４的表达对ＡＭＬ－Ｍ４、Ｍ５尤其是Ｍ５亚型的鉴别诊断有重要价值。     

分化诱导剂联合雄激素或小剂量化疗药物治疗骨髓增生异常综合征

报道３６例骨髓增生异常综合征（ＭＤＳ）的治疗效果。原始细胞不增多的ＭＤＳ（ＲＡ和ＲＡＳ）１７例，以雄激素丙酸睾丸素或康力龙联合分化诱导剂全反式维甲酸和１，２５（ＯＨ）２Ｄ３治疗，部分病例尚加用小剂量强的松；原始细胞增多的ＭＤＳ（ＲＡＥＢ、ＣＭＭＬ及ＲＡＥＢＴ）１９例，以小剂量阿糖胞苷或／和小剂量阿克拉霉素化疗并联用分化诱导剂。前组的有效率为７０．６％，基本缓解率为２９．４％，后组的有效率为５７．９％，完全缓解（ＣＲ）率为３６．８％，两组的总有效率为６３．９％。５例取得基本缓解的ＲＡ和ＲＡＳ目前都处于缓解期，已６个月～１６个月；７例ＣＲ的ＲＡＥＢ和ＲＡＥＢＴ３例于ＣＲ后６个月～１２个月转为急性白血病，另４例仍处于ＣＲ中，已３个月～５２个月。认为根据ＭＤＳ患者的类型予不同的药物联合治疗可取得较好的疗效。     

100例急性白血病患者多药耐药性检测分析

作者应用抗Ｐ－糖蛋白（Ｐ－１７０）单抗ＪＳＢ－１对１００例急性白血病（ＡＬ）患者进行了检测，结果表明：（１）包括Ｍ３在内的几乎所有ＡＬ亚型均可有ＭＤＲ阳性表达，初诊组ＭＤＲ表达率以Ｍ５（６２．５％）为最高，ＡＮＬＬ（２０％）虽高于ＡＬＬ（８．３％），但差异无显著性意义。（２）初诊组部分患者（１８．２％）呈ＭＤＲ阳性表达，提示ＭＤＲ的内在性。（３）Ｐ－１７０阳性率与ＡＬ临床状况密切相关，难治组Ｐ－１     

白血病化疗药物体外敏感性检测与临床疗效的分析

 19例急性白血病，用核仁组成区相关嗜银蛋白（AgNOR）检测技术，测定白血病细胞对化疗药物体外药敏。参考药敏度选择化疗药物行体内化疗。结果显示：体外药敏与体内疗效总符合率为：81.8%，阳性符合率为79.2%，阴性符合率为85.7%，敏感度为95.0%，特异性为61.5%。临床治疗效果，ANLL明显优于ALL，完全缓解率分别为84.6%，33.3%。该法简便、快速、敏感，对急性白血病的化疗有重要参考价值。     

急性白血病化疗药物敏感性ATP法的临床应用

 用ATP法对26例白血病病人进行体外药敏检查，观察其中18例急性白血病病人体外药敏与体内疗效关系。急性淋巴细胞白血病4例、急性非淋巴细胞白血病11例、慢性粒细胞白血病急性变3例。总符合率为83.3%，阳性符合率91.7%，阴性符合率66.7%.结果提示ATP法具有定量、短期、客观的临床应用价值。     

Chemosensitivity in vitro to 2-chlorodeoxyadenosine and 9-beta-D-arabinofuranosyl-2-fluoroadenine in previously unexposed cases of chronic lymphocytic leukemia

The chemosensitivity of leukemia cells, obtained from the peripheral blood of 35 patients with B-cell chronic lymphocytic leukemia, was determined by a leucine-incorporation assay in vitro. There was good correlation between the sensitivities to two purine analogs, 2-chlorodeoxyadenosine and 9-beta-D-arabinofuranosyl-2-fluoroadenine among previously untreated patients when tested at the 80% inhibition level. Previous exposure to chlorambucil did not affect the sensitivity to these compounds.     

High in vitro-in vivo correlation of drug response using sponge-gel-supported three-dimensional histoculture and the MTT end point.

The in vitro sponge-gel-supported three-dimensional histoculture chemosensitivity assay (Hoffman assay) allows the in vivo-like culture of human tumors. In this study, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H tetrazolium bromide (MTT) end point was applied to the Hoffman assay in an attempt to increase in vitro-in vivo correlation. The chemosensitivities of 16 human tumor lines were determined in vitro by the histoculture assay, and retrospectively correlated to their in vivo chemosensitivity as xenografts in nude mice. The in vitro test was considered to be positive if tumor-cell MTT reduction activity was lowered by more than 50%. The cutoff drug concentrations to determine sensitivity in vitro were determined for mitomycin C, doxorubicin, 5-fluorouracil and cisplatin. Using these cutoff drug concentrations in vitro we found, as a function of time of exposure, a strong correlation between serum drug concentrations found in nude mice given maximum tolerated doses and drug concentrations found in the histoculture media in vitro, thereby establishing a relationship between the amounts of drugs to which tumors were exposed in vivo and in vitro. The overall correlation rate of the efficacy results of the drug-response assay to in vivo chemosensitivities was 89.8%, with 90.0% true-positive and 89.7% true-negative rates, 81.7% sensitivity and 94.6% specificity, thereby indicating potential clinical use for tumor histoculture with the MTT end point.     

Clinical correlations with chemosensitivities measured in a simplified tritiated thymidine incorporation assay in patients with malignant effusion

Utilizing a simplified tritiated thymidine incorporation assay, in vitro chemosensitivity of tumor cells obtained from malignant effusions was assessed and, in these patients, chemotherapeutic drugs were administered directly into the peritoneal or pleural cavity. Then, correlations between in vitro sensitivity and clinical response were investigated. Fifteen (88%) of 17 patients with various carcinomas gave evaluable chemosensitivity results. All 15 patients were evaluable for in vitro-in vivo correlations. This assay had an accuracy of 75% for prediction of response (3 of 4) and an accuracy of 82% for prediction of resistance (9 of 11), when the peak achievable plasma concentrations were selected as the concentrations used for in vitro sensitivity testing, adopting 80% inhibition of thymidine incorporation as cutoff level.     

In vitro sensitivity of human melanoma xenografts to cytotoxic drugs. Correlation with in vivo chemosensitivity

Single-cell suspensions prepared from five human melanomas, grown serially as xenografts in athymic nude mice, were exposed in vitro to increasing concentrations of DTIC (Dacarbazine), CCNU (Lomustine), procarbazine, vinblastine, and the cancerostatic lectins abrin and ricin. The in vitro chemosensitivity of the cells, as measured by the drug concentrations required to inhibit colony formation in soft agar by 50%, was correlated with the growth delay of the xenografts in vivo, previously observed after treatment of the animals with maximum tolerable doses of the same drugs. It was found that for each drug the in vitro sensitivity of the different xenografts was strongly correlated with their response in vivo. The results provide evidence that the soft agar test, as carried out here, adequately reflects the relative sensitivity of the xenografts in vivo. The data indicate that human xenografts may be used to develop quantitative in vitro chemosensitivity tests.     

Prediction of Chemotherapeutic Response in Unresectable Non-small-cell Lung Cancer (NSCLC) Patients by 3-(4,5-Dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) Assay

Background: Selecting chemotherapy regimens guided by chemosensitivity tests can provide individualizedtherapies for cancer patients. The 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2Htetrazolium,inner salt (MTS) assay is one in vitro assay which has become widely used to evaluate the sensitivityto anticancer agents. The aim of this study was to evaluate the clinical applicability and accuracy of MTS assayfor predicting chemotherapeutic response in unresectable NSCLC patients. Methods: Cancer cells were isolatedfrom malignant pleural effusions of patients by density gradient centrifugation, and their sensitivity to eightchemotherapeutic agents was examined by MTS assay and compared with clinical response. Results: A totalof 37 patients participated in this study, and MTS assay produced results successfully in 34 patients (91.9%).The sensitivity rates ranged from 8.8% to 88.2%. Twenty-four of 34 patients who received chemotherapy wereevaluated for in vitro-in vivo response analysis. The correlation between in vitro chemosensitivity result and invivo response was highly significant (P=0.003), and the total predictive accuracy, sensitivity, specificity, positivepredictive value, and negative predictive value for MTS assay were 87.5%, 94.1%, 71.4%, 88.9%, and 83.3%,respectively. The in vitro sensitivity for CDDP also showed a significant correlation with in vivo response (P=0.018,r=0.522). Conclusion: MTS assay is a preferable in vitro chemosensitivity assay that could be use to predict theresponse to chemotherapy and select the appropriate chemotherapy regimens for unresectable NSCLC patients,which could greatly improve therapeutic efficacy and reduce unnecessary adverse effects.     

The clinical application of an ATP assay to predict the chemosensitivity of gastric cancer

The chemosensitivity of human xenografts in nude mice and fresh surgical specimens of gastric cancer was evaluated in vitro using the ATP assay and the MTT assay. The in vitro sensitivity of 6 human xenografts was detected by the ATP assay and compared with the in vivo sensitivity of the xenografts in nude mouse. The assay showed a 56.3% true-positive rate and a 85.7% true-negative rate, with 90.0% sensitivity and 46.2% specificity. When 10 surgical specimens obtained from gastric cancer patients were divided into two groups and sensitivities assessed by the ATP and the MTT assays, the overall correlation of both assays was 81.3%. The ATP assay might be useful in evaluating the chemosensitivity of human gastric carcinomas.     

In vitro chemosensitivity test using collagen gel matrix for human gastric carcinomas]

We evaluated the usefulness of in vitro human tumor culture system using a specialized collagen gel matrix derived from pigskin as a chemosensitivity test for human gastric cancer including scirrhous type. Seven xenograft tumors derived from human gastric cancers were examined with this system and compared with the data obtained by the nude mouse chemosensitivity assay. Xenograft tumors exhibited an in vivo like three-dimensional growth on the collagen gel matrix. There were very close associations between the results obtained by this assay and those obtained from the same drug by the nude mouse assay. Moreover, this system seemed to be useful for prediction of drug sensitivity for scirrhous type gastric cancers. This in vitro assay system has an advantage for chemosensitivity test, because of its convenience, rapidity, and in vivo like three-dimensional tumor growth.     

Changes in chemosensitivity of K 562 leukemia cells after induction of erythroid differentiation by hemin

The human leukemia cell line K 562, when treated with subcytotoxic doses of hemin, undergoes reversible erythroid commitment, as shown by the increased synthesis of hemoglobin. Hemin-treated cells maintain replicative capabilities, although perturbations in cell cycle kinetics are induced. K 562 cells were used to investigate changes in antitumor drug sensitivity as a consequence of cell differentiation induced by hemin treatment. K 562 leukemia cells, cultured in the presence of 20 microM hemin for 12 days, were treated with non-phase-specific (adriamycin, 4-OOH-cyclophosphamide, mitomycin C, bleomycin, cis-diamminedichloro platinum) and phase-specific (vincristine, methotrexate and 5-fluorouracil) antitumor drugs. The results obtained by chemosensitivity tests showed a generalized decrease in chemosensitivity of the K 562 cells to all the drugs tested as a consequence of the hemin-induced differentiation.     

Colorimetric chemosensitivity testing using sulforhodamine B

A colorimetric chemosensitivity test was investigated using sulforhodamine B (SRB), which stains protein synthesized by cells, as an end-point marker. Four cultured cell lines, 9 human tumor xenografts serially transplanted into nude mice, and 14 fresh surgical specimens were subjected to this assay. The optimal conditions for the assay were 3–5 × 10⁴ cells per well in a 96-microplate, an SRB concentration of 4%, and an incubation time of more than 10 minutes. When mitomycin C, doxorubicin, cisplatin, and 5-fluorouracil were assessed by the SRB assay, the concentration-effect curves revealed a sharp slope between plateaux at low and high concentrations, suggesting that this assay has an excellent sensitivity which can assess the effect of drugs as “all or none.” Although this high sensitivity resulted in good reproducibility of the assay for cultured cell lines, the predictive rate of the SRB assay for the chemosensitivity of human tumor xenografts in vivo was limited to 63.9%. As a result, this SRB assay is thought to be useful for evaluating the chemosensitivity of cultured cells as all or none, since it can assess directly cellular protein synthesis, which is one of the most important parameters of cell renewal, with excellent sensitivity. © 1993 Wiley-Liss, Inc.