Rapid and reliable detection of CD44 variants in gastric carcinoma using a nested reverse transcription-polymerase chain reaction.

Abstract

The present study aimed to establish a rapid and reliable method for detecting the expression of cluster of differentiation 44 variant (CD44v) in gastric carcinoma, and to investigate the significance of CD44v in gastric carcinoma. Using a nested reverse transcription-polymerase chain reaction (RT-PCR) technique, the expression of CD44v and CD44v8-10 was analyzed in gastric cancer tissues (128 cases), precancerous lesions (19 cases of atypical hyperplasia and 6 cases of intestinal metaplasia) and corresponding adjacent non-cancerous tissues (153 cases). The tumor and non-cancerous biopsy samples of 153 patients were analyzed using nested RT-PCR. All the PCR products included bands at 482 bp, demonstrating positive CD44 expression. By contrast, the CD44v band (>600 bp) was observed in 132/153 total tumor samples (86.3%), including 114/128 gastric cancer samples (89.1%), 16/19 atypical hyperplasia samples (84.2%) and 2/6 intestinal metaplasia samples (33.3%). However, 18/153 non-cancerous tissues samples (11.8%) exhibited a CD44v band. Thus, CD44v expression was significantly higher in gastric cancer tissues and precancerous lesions compared with that of adjacent non-cancerous tissues (P<0.05). Furthermore, there was a significant difference in CD44v8-10 expression detected between gastric cancer and adjacent non-cancerous tissue samples (P<0.05). Among the 25 patients with precancerous lesions, 8/19 atypical hyperplasia cases and 1/6 intestinal metaplasia cases were positive for CD44v8-10 expression. The difference in the CD44v8-10 expression rate among the various pathological types of gastric cancer (n=128) cases was not significant (P>0.05). Additionally, immunohistochemical analysis identified CD44v positivity (++) in 59/76 (77.6%) cases of gastric cancer and 5/12 (41.1%) cases of atypical hyperplasia. The CD44v and CD44v8-10 PCR products were confirmed by sequencing analysis. The results of the present study indicated that nested RT-PCR technology may be exploited as a method for gastric carcinoma diagnosis.