Molecular diagnosis of exocrine pancreatic cancer using a percutaneous technique

被引:23
作者
Evans, DB
Frazier, ML
Charnsangavej, C
Katz, RL
Larry, L
Abbruzzese, JL
机构
[1] UNIV TEXAS, MD ANDERSON CANC CTR, DEPT GASTROINTESTINAL MED ONCOL & DIGEST DIS, HOUSTON, TX 77030 USA
[2] UNIV TEXAS, MD ANDERSON CANC CTR, DEPT DIAGNOST RADIOL, HOUSTON, TX 77030 USA
[3] UNIV TEXAS, MD ANDERSON CANC CTR, DEPT PATHOL, HOUSTON, TX 77030 USA
关键词
pancreatic cancer; K-ras; FNA; diagnosis;
D O I
10.1007/BF02306278
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
Background: The K-ras oncogene is activated by point mutations at codon 12 in most patients with exocrine pancreatic cancer, Mutant-enriched polymerase chain reaction (PCR) amplification can enhance the detection of mutated K-ras, This technique was applied to patients undergoing percutaneous fine-needle aspiration (FNA) biopsy of suspect pancreatic lesions. Methods: Twenty-five patients underwent percutaneous FNA of the pancreas for cytologic and molecular analysis. After preparing cytologic smears, the 22-gauge needle and syringe used for FNA were rinsed in RPMI-1640, The specimen was centrifuged, and DNA was extracted from the supernatant and subjected to mutant-enriched PCR using appropriate mismatched primers that introduce a BstNI restriction endonuclease clevage site at codon 12 of wild-type, but not mutant, K-ras. After digestion with BstNI, the DNA was reamplified. To increase assay sensitivity, the final five PCR cycles were completed incorporating 5 mu Ci of (alpha-32P)dCTP, The DNA was then redigested and subjected to gel electrophoresis and autoradiography. Results: The median amount of DNA retrieved per specimen was 3.33 mu g. Mutant Kras was detected as a band of 143 bps; residual wild-type DNA was seen as a 114-bp fragment, Twenty-one of 25 specimens demonstrated mutated K-ras DNA. Two patients with nondiagnostic cytology results had mutated K-ras DNA; adenocarcinoma of pancreatic origin was confirmed in both cases after pancreatectomy. Conclusion: The molecular diagnosis of pancreatic cancer through identification of mutations in K-ras can be readily performed on specimens obtained by percutaneous FNA. As aggressive multimodality management of this disease becomes more common, pretreatment analysis of molecular determinants may have greater clinical significance.
引用
收藏
页码:241 / 246
页数:6
相关论文
共 51 条
[1]  
Abbruzzese James L., 1994, Cancer Bulletin (Houston), V46, P525
[2]  
ABBRUZZESE JL, 1994, INT J PANCREATOL, V16, P331
[3]   PANCREATIC-CANCER CELL-DNA CONTENT CORRELATES WITH LONG-TERM SURVIVAL AFTER PANCREATICODUODENECTOMY [J].
ALLISON, DC ;
BOSE, KK ;
HRUBAN, RH ;
PIANTADOSI, S ;
DOOLEY, WC ;
BOITNOTT, JK ;
CAMERON, JL .
ANNALS OF SURGERY, 1991, 214 (06) :648-656
[4]   MOST HUMAN CARCINOMAS OF THE EXOCRINE PANCREAS CONTAIN MUTANT C-K-RAS GENES [J].
ALMOGUERA, C ;
SHIBATA, D ;
FORRESTER, K ;
MARTIN, J ;
ARNHEIM, N ;
PERUCHO, M .
CELL, 1988, 53 (04) :549-554
[5]   RANDOMIZED TRIAL OF ENDOSCOPIC ENDOPROSTHESIS VERSUS OPERATIVE BYPASS IN MALIGNANT OBSTRUCTIVE-JAUNDICE [J].
ANDERSEN, JR ;
SORENSEN, SM ;
KRUSE, A ;
ROKKJAER, M ;
MATZEN, P .
GUT, 1989, 30 (08) :1132-1135
[6]  
[Anonymous], 1979, Ann Surg, V189, P205
[7]   COMPARATIVE-ANALYSIS OF MUTATIONS IN THE P53 AND K-RAS GENES IN PANCREATIC-CANCER [J].
BERROZPE, G ;
SCHAEFFER, J ;
PEINADO, MA ;
REAL, FX ;
PERUCHO, M .
INTERNATIONAL JOURNAL OF CANCER, 1994, 58 (02) :185-191
[8]  
CALDAS C, 1994, CANCER RES, V54, P3568
[9]  
CAMERON JL, AM J SURG, V191, P161
[10]  
EVANS DB, 1992, ARCH SURG-CHICAGO, V127, P1335