Combined DNA flow cytometry and sorting with k-ras2 mutation spectrum analysis and the prognosis of human sporadic colorectal cancer

被引:23
作者
Geido, E
Sciutto, A
Rubagotti, A
Oliani, C
Monaco, R
Risio, M
Giaretti, W
机构
[1] Natl Inst Canc Res, Lab Biophys & Cytometry, I-16132 Genoa, Italy
[2] Univ Genoa, Ctr Biostat, Genoa, Italy
[3] Natl Inst Canc Res, Ist Nazl Ric Canc, I-16132 Genoa, Italy
[4] Osped Civile, Div Med Oncol, I-37126 Verona, Italy
[5] Osped Antonio Cardarelli, Serv Anat & Istol Patol, Naples, Italy
[6] Ist Ric & Cura Cancro, Serv Anat & Istol Patol, Candiolo, Italy
来源
CYTOMETRY | 2002年 / 50卷 / 04期
关键词
chromosome instability; aneuploidy; oncogenes; tumor progression;
D O I
10.1002/cyto.10109
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Background: Activation of the k-ras2 pathways and chromosomal instability leading to aneuploidy in human sporadic colorectal cancer (sCRC) is essential to the tumor cell ability to survive, grow, and metastatize. Methods: The study included 135 patients with sCRC who were followed up for a median of 72 months. Multiple fresh-frozen fragments obtained from superficial and invasive areas of the tumors were mixed and used to detect the degree of DNA aneuploidy (DNA index [DI]) and S-phase fraction by two scatter signals and 4,6-diamidino-2-phenylindole-2-hydrocioride (DAPI) fluorescence flow cytometry (FCM). PCR amplification and k-ras2 mutation spectrum analysis were performed using enriched epithelial nuclei after sorting DNA aneuploid nuclei and DNA diploid nuclei from which tissue-infiltrating lymphocytes were absent. Results: DNA aneuploidy was detected in 98 (73%) and k-ras2 mutations in 54 cases (40%). Univariate analyses of overall survival with both Dukes' A to D or B to C series of cases showed that DNA multiple aneuploidy, k-ras2 mutations, older age, and distal site, but not increased S-phase fraction, were predictive of worse outcome. Multivariate Cox models strongly indicated that k-ras2 mutations, but neither single nor multiple DNA aneuploidy, were an independent prognostic factor in both series of patients. In particular, with B and C Dukes' stage patients (n = 110), the relative risk (RR) of death was above 2.5 with k-ras2 mutations and above 3 with the G-->C/T transversions. Conclusion: Combined FCM and k-ras2 analysis may be used to predict long-term increased risk of death in sCRC patients. (C) 2002 Wiley-Liss, Inc.
引用
收藏
页码:216 / 224
页数:9
相关论文
共 39 条
[1]  
Ahnen DJ, 1998, CANCER RES, V58, P1149
[2]   Ras links growth factor signaling to the cell cycle machinery via regulation of cyclin D1 and the Cdk inhibitor p27(KIP1) [J].
Aktas, H ;
Cai, H ;
Cooper, GM .
MOLECULAR AND CELLULAR BIOLOGY, 1997, 17 (07) :3850-3857
[3]   Kirsten ras mutations in patients with colorectal cancer: the multicenter "RASCAL" study [J].
Andreyev, HJN ;
Norman, AR ;
Cunningham, D ;
Oates, JR ;
Clarke, PA .
JOURNAL OF THE NATIONAL CANCER INSTITUTE, 1998, 90 (09) :675-684
[4]  
[Anonymous], 1989, SYNTHETIC OLIGONUCLE
[5]  
BAKER SJ, 1990, CANCER RES, V50, P7717
[6]  
BARDI G, 1993, CANCER RES, V53, P1895
[7]  
Bast RC, 1996, J CLIN ONCOL, V14, P2843
[8]   CONSENSUS REVIEW OF THE CLINICAL UTILITY OF DNA FLOW-CYTOMETRY IN COLORECTAL-CANCER [J].
BAUER, KD ;
BAGWELL, CB ;
GIARETTI, W ;
MELAMED, M ;
ZARBO, RJ ;
WITZIG, TE ;
RABINOVITCH, PS .
CYTOMETRY, 1993, 14 (05) :486-491
[9]   PREVALENCE OF RAS GENE-MUTATIONS IN HUMAN COLORECTAL CANCERS [J].
BOS, JL ;
FEARON, ER ;
HAMILTON, SR ;
VERLAANDEVRIES, M ;
VANBOOM, JH ;
VANDEREB, AJ ;
VOGELSTEIN, B .
NATURE, 1987, 327 (6120) :293-297
[10]   Carcinogenesis and natural selection: A new perspective to the genetics and epigenetics of colorectal cancer [J].
Breivik, J ;
Gaudernack, G .
ADVANCES IN CANCER RESEARCH, VOL 76, 1999, 76 :187-+