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Molecular Analysis of Single Tumor Glands Using the Crypt Isolation Method in Endometrial Carcinomas
  1. Takayuki Nagasawa, MD, PhD,
  2. Tamotsu Sugai, MD, PhD,
  3. Tadahiro Shoji, MD, PhD,
  4. Wataru Habano, PhD and
  5. Toru Sugiyama, MD, PhD
  1. * Departments of Obstetrics and Gynecology,
  2. Molecular Diagnostic Pathology, and
  3. Pharmacodynamics and Molecular Genetics, School of Medicine, Iwate Medical University, Morioka, Japan.
  1. Address correspondence and reprint requests to Tamotsu Sugai, MD, PhD, Division of Molecular Diagnostic Pathology, Iwate Medical University, 19-1 Uchimaru, Morioka City, Iwate, 020-8505, Japan. E-mail: tsugai{at}


Objective Endometrial adenocarcinomas are characterized by the presence of many single tumor glands in which multiple genetic changes have accumulated. To elucidate the differences in molecular abnormalities among single tumor glands, individual tumor glands were analyzed and microsatellite alterations (loss of heterozygosity (LOH) and microsatellite instability [MSI]) were examined using the crypt isolation method in glands from each tumor from patients with endometrial carcinoma.

Methods Twenty-five patients with endometrial adenocarcinoma who underwent surgery were included in this study. We obtained cancerous individual isolated tumor glands from each patient using the crypt isolation method. For LOH and MSI analyses, we used 15 microsatellite markers (3p, 5q, 10q, 13q, 17p, 18q, BAT25, and BAT26) and the promoter regions of 6 genes (transforming growth factor beta receptor II, BAX, insulin-like growth factor II receptor, E2F4, MutS homolog 3, and MSH6).

Results Loss of heterozygosity was detected in 8 (32%) of 25 patients, and MSI was detected in 9 (36%) of 25 patients. Some MSI-positive carcinomas had LOH in single tumor gland samples, and the coexistence of LOH and MSI was confirmed. In 16 (64%) of 25 cases, intratumoral genetic heterogeneity among single tumor gland samples was detected.

Conclusions By analyzing multiple single tumor glands within the same tumor, we found that endometrial adenocarcinoma was composed of various tumor glands with different molecular abnormalities, even in a limited region within the same tumor.

  • Endometrial adenocarcinoma
  • Crypt isolation method
  • Cost of heterozygosity
  • Microsatellite instability

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  • The authors declare no conflicts of interest.