Basic study of in vitro BrdU labeling method as a chemosensitivity test.
Grant-in-Aid for General Scientific Research (C)
|Allocation Type||Single-year Grants|
|Research Institution||Yamaguchi University, School of Medicine|
SHIMABUKURO Tomoyuki(1992) Yamaguchi University, Research School of Med.Hospital Associate, 医学部附属病院, 助手 (60226222)
山本 憲男(1990-1991) 山口大学, 医学部, 助教授 (10034985)
NAITO Katsusuke Yamaguchi University, Professor School of Medicine, 医学部, 教授 (60115251)
MURAKAMI Tomoyuki Yamaguchi University, Assistant School of Medicine Professor, 医学部, 講師 (20200272)
MATSUYAMA Hideyasu Yamaguchi University, Research School of Med. Hospital Associate, 医学部附属病院, 助手 (70209667)
島袋 智之 山口大学, 医学部・附属病院, 助手 (60226222)
|Project Period (FY)
1990 – 1992
Completed(Fiscal Year 1992)
|Budget Amount *help
¥1,300,000 (Direct Cost : ¥1,300,000)
Fiscal Year 1992 : ¥600,000 (Direct Cost : ¥600,000)
Fiscal Year 1991 : ¥700,000 (Direct Cost : ¥700,000)
|Keywords||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|
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 method
We 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
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 rate
Our 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
Research Output (18results)