Individual mRNA species from normal and neoplastic endometrium obtained from the same patient were comparatively studied by exploiting the "differential display" methodology. The mRNAs were extracted from tissues, reverse transcribed, and amplified by PCR using appropriate primers. The cDNA electrophoretic bands which were frankly different on the basis of their quantitative expression were excised from the gels and reamplified. Each of these sequences was subsequently screened for the capacity to hybridize to RNA preparations from normal endometrium and endometrial adenocarcinoma samples, first from the original patient and then from a group of selected patients. Of many, only two sequences, thereafter named N5.5 and T16.2, respectively, successfully passed the two sieving tests and were chosen for further analysis. It appears that N5.5 recognizes an mRNA which is expressed more abundantly in normal than in neoplastic (adenocarcinoma) endometrium; in contrast, T16.2 seems to preferentially recognize an mRNA which is expressed in tumoral tissues. These two sequences have been cloned and sequenced; they do not show any identity or significant similarity to any other known sequence.
Identification of differentially expressed mRNAs in normal and neoplastic (adenocarcinoma) human endometrium.
TECCE, Mario Felice;
1996-01-01
Abstract
Individual mRNA species from normal and neoplastic endometrium obtained from the same patient were comparatively studied by exploiting the "differential display" methodology. The mRNAs were extracted from tissues, reverse transcribed, and amplified by PCR using appropriate primers. The cDNA electrophoretic bands which were frankly different on the basis of their quantitative expression were excised from the gels and reamplified. Each of these sequences was subsequently screened for the capacity to hybridize to RNA preparations from normal endometrium and endometrial adenocarcinoma samples, first from the original patient and then from a group of selected patients. Of many, only two sequences, thereafter named N5.5 and T16.2, respectively, successfully passed the two sieving tests and were chosen for further analysis. It appears that N5.5 recognizes an mRNA which is expressed more abundantly in normal than in neoplastic (adenocarcinoma) endometrium; in contrast, T16.2 seems to preferentially recognize an mRNA which is expressed in tumoral tissues. These two sequences have been cloned and sequenced; they do not show any identity or significant similarity to any other known sequence.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
