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Basic study of in vitro BrdU labeling method as a chemosensitivity test.

体外 BrdU 标记方法作为化学敏感性试验的基础研究。

基本信息

批准号：
02670709
负责人：
SHIMABUKURO Tomoyuki
金额：
$ 0.83万
依托单位：
Yamaguchi University, School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1992
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02670709/
关键词：
In vitro BrdU labeling method Chemosensitivity test PCNA Ki-67 AgNOR flow cytometry Cell kinetics Intratumoral Pt concentration 抗BrDU抗体抗PCNA抗体 In vitro BrDU標識細胞動態

项目摘要

It is very important to determine the anticancer drug sensitivity of individual cancers in order to increase the effects and decrease the side effects. To this end, we have studied a new chemosensitivity test using in vitro and in vivo BrdU labeling method.1.In vitro brdU labeling methodWe investigated a new chemosensitivity test by analyzing percent inhibition rate of BrdU labeled S-phase cells using immunohistochemical staining technique. Three anticancer drugs, adriamycin(ADM), cis-platin(CDDP), and vincristin(VCR) were tested using the MBT-2 solid tumor originated from FANFT-induced mouse bladder cell line. The anticancer effects evaluated by this method were compared with corresponding percent colony survival rate by colony assay.The labeling indices of BrdU were stable within four hours after tumor excision. The inhibition rate of BrdU-uptake of ADM was influenced by drug concentration, but that of VCR did not correspond drug concentration. Whether that of CDDP depended on drug c … More oncentration and exposure time.This method may be able to apply to a new chemosensitivity test, because the curves of percent colony survivals were similar to that of the inhibition rate of BrdU-uptake.2.In vivo BrdU labeling method.The CoH mice with subcutaneous MBT-2 tumor were injected intraperitoneally of 1,1/2,1/5,1/10 of LD_<10> dose, respectively. The anticancer effects of CDDP were evaluated by measuring the tumor volume every day for 3 weeks. The obtained results were compared to the % inhibition rate of BrdU labeled cells or PC NA positive cells. Our preliminary data show that with further research in this fields we will be able to determine the effectiveness of chemotherapy in clinical pratice also.3.Clinical usefulness of PCNA positive rateOur preliminary clinical data in 12 patients with bladder carcinoma show that PCNA index is able to determine the effectiveness of chemotherapy. PCNA index was calculated in biopsy materials before and after 3 courses of intra-arterial chemotherapy. PCNA positive rate was 41,4*4,4%(mean*SE) before and 20.4*4.3% after the chemotherapy, respectively. Four out of 12 patients had recurrence of the tumor and in them repeat biopsy material showed higher PCNA positive rate than at the beginning of the intra-arterial chemotherapy. Our data suggested that this method may be applied for a new chemosensitivity test. Less

确定个体癌症的抗癌药物敏感性对于增加疗效和减少副作用是非常重要的。为此，我们研究了一种新的体外和体内BrdU标记的药物敏感性试验方法。1.体外BrdU标记方法我们研究了一种新的药物敏感性试验，通过免疫组织化学染色技术分析BrdU标记的S期细胞的抑制率。三种抗癌药物，阿霉素（ADM），顺铂（CDDP），长春新碱（VCR）进行了测试使用的MBT-2实体瘤来源于FANFT诱导的小鼠膀胱细胞系。将该方法评价的抗癌效果与集落测定相应的集落存活率进行比较，肿瘤切除后4小时内BrdU标记指数稳定。ADM对BrdU摄取的抑制率受药物浓度的影响，而VCR对BrdU摄取的抑制率与药物浓度无关。CDDP的疗效是否依赖于药物c ...更多信息 2.体内BrdU标记法：将CoH小鼠皮下接种MBT-2肿瘤，分别以LD_（1）、LD_（1/2）、LD_（1/5）、LD_（1/10）剂量腹腔注射<10>。通过每天测量肿瘤体积来评估CDDP的抗癌效果，持续3周。将获得的结果与BrdU标记的细胞或PCNA阳性细胞的%抑制率进行比较。我们的初步资料表明，随着这一领域的进一步研究，我们将能够在临床实践中判断化疗的有效性。3. PCNA阳性率的临床应用我们对12例膀胱癌患者的初步临床资料表明，PCNA指数能够判断化疗的有效性。分别于化疗前、化疗3个疗程后取活检标本，计算PCNA指数。PCNA阳性率化疗前为41.4 * 4.4%（均数），化疗后为20.4*4.3%（均数）。12例患者中有4例肿瘤复发，其中重复活检材料显示PCNA阳性率高于动脉化疗开始时。本研究结果提示，该方法可作为一种新的药物敏感性试验方法。少