Abstract
In order to identify the possible role of the DCC gene in neoplasms of the human female reproductive tract, messenger RNA expression of the DCC gene was examined by reverse transcriptase-polymerase chain reaction, and expression of the DCC gene product was detected immunohistochemically. While histologically normal endometrium, cervical epithelium and ovary expressed detectable mRNA of the DCC gene, three of eight (37%) endometrial carcinomas, one of two (50%) cervical carcinomas and 9 of 22 (41%) ovarian malignant tumours had significantly reduced or negligible DCC expression, and another endometrial carcinoma and two other ovarian tumors underexpressed DCC when compared with histologically normal endometrial or ovarian tissues. Impaired DCC mRNA expression was detected more frequently in grade 3 ovarian epithelial tumours than in grade 1 tumours (P = 0.002). Loss of expression of the DCC gene product detected by immunohistochemistry significantly correlated with the loss of mRNA expression in ovarian carcinomas (P = 0.01 by chi-square test) or in both endometrial and ovarian carcinomas combined (P = 0.001). Loss of heterozygosity of the DCC gene was also evaluated by restriction fragment polymorphism analysis of the polymerase chain reaction-amplified DNA fragment. Loss of heterozygosity of the DCC gene was detected in one of seven (14%) informative cases of endometrial carcinomas, 1 of 11 (9%) informative cases of cervical carcinomas and two of six (33%) informative cases of ovarian tumours. These results demonstrate that inactivation of the DCC gene, especially by the loss of expression, plays a significant role in the aetiology of neoplasms of the human reproductive tract.
Highlights
We report here that the inactivation of DCC occurs rather frequently in these neoplasms
A monohistopathological diagnosis and the remainder were quickly clonal antibody to the DCC gene product (Ab-1) (Oncogene frozen for extraction of RNA or DNA or for immunohisto- Science, Uniondale, NY, USA) was used as primary antibody chemical analysis
A total of 32 neoplasms, which normal mouse serum were incubated instead of the including eight endometrial adenocarcinomas, 22 ovarian corresponding primary antibody were used as negative conmalignant and low-grade malignant tumours
Summary
DCC gene product was detected immunohistochemically in a PCR amplification was performed to generate 367 bp DNA total of 18 neoplasms, including in five endometrial adeno- fragments around the polymorphic MspI sites of the DCC carcinomas and 13 malignant ovarian tumours PCR amplification was performed from the deoxynucleotide triphosphate, 0.5 U of Taq polymerase complementary DNA to generate the 233 bp fragment of the (Perkin-Elmer Cetus, Norwalk, CT, USA), 1.5 mM magne- DCC gene and the 319 bp fragment of A-actin simultaneoussium chloride, 50 mM potassium chloride, 1O mM Tris-HCI ly. Both fragments were successfully amplified in the his-. Lanes [1,2,3,4,5], endometrial carcinomas, lanes 6 and 7. cervical carcinomas: lanes 8 -15. ovarian carcinomas; lanes 16- 18. normal uterine endometrium. normal ovary, and normal colon mucosa
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