Identification of differentially expressed genes in esophageal cancer through SSH in combination with high throughput reverse Northern screening |
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Authors: | Jin Zhou Zhihua Liu Xiuqin Wang Fang Ding Chuannong Zhou Min Wu |
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Institution: | (1) National Laboratory of Molecular Oncology, Department of Cell Biology, Cancer Institute, Peking Union Medical College and Chinese Academy of Medical Sciences, 100021 Beijing, China |
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Abstract: | To understand the molecular mechanisms of carcinogenesis of esophagus and to isolate genes with different expression levels
in esophageal cancer, suppression subtractive hybridization (SSH) was combined with PCR-based cDNA synthesis and reverse Northern
on the cancer tissues and matched almost normal mucosa using 5 microgram of total RNA as starting marterial. Eight genes were
found expressed differentially in esophageal cancer, in which 5 were known genes and 3 were novel ones; and 6 were down-regulated
in cancer tissues, while 2 were up-regulated; 6 were of mid-high abundance and 2 were of low abundance in esophagus. The results
revealed that alteration in expression level of multiple genes underlied the initiation and development of esophageal cancer.
The differentially expressed genes identified in this study such asliporcotin I,cystalin A, cystatin B, cytokeratin 13 may play roles in dedifferentiation, transformation and malignant proliferation of esophageal cancer. The combination of
SSH with PCR-based doublestrand cDNA synthesis and high throughput reverse Northern screening is an efficient way to isolate
differentially expressed genes from microgram of total RNA. |
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Keywords: | esophageal cancer suppression subtractive hybridization (SSH) differential expression lipocortin cystatin |
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