ORIGINAL ARTICLE

Detection of genes expressed in primary colon cancers by in situ hybridisation: overexpression of RACK 1

A Saito¹, G Fujii¹, Y Sato³, M Gotoh¹, M Sakamoto¹, G Toda² and S Hirohashi¹

¹ Pathology Division, National Cancer Center Research Institute, 1–1 Tsukiji, 5-chome, Chuo-ku, Tokyo, 104–0045, Japan
² Gastroenterology and Hepatology Division, Department of Internal Medicine, Jikei University School of Medicine, Tokyo, 105-8461, Japan
³ Department of Pathology, School of Allied Health Sciences, Kitasato University, Kanagawa, 228-8555, Japan

Correspondence to:
Correspondence to:
Dr S Hirohashi, Pathology Division, National Cancer Center Research Institute, 1–1 Tsukiji, 5-chome, Chuo-ku, Tokyo, 104–0045, Japan;
shirohas{at}ncc.go.jp

Aims: The isolation of various genes that are expressed in a region specific manner is considered useful for research in molecular pathology. In situ hybridisation (ISH) was used in a screening procedure to isolate these genes efficiently, using colon cancer as a model.

Methods: Suppression subtractive hybridisation (SSH) between colon cancer tissue samples and corresponding non-cancerous tissues was performed. Genes showing high expression in the cancers were selected using macro-DNA array analysis. As a final screening procedure, conventional ISH was performed to isolate genes expressed specifically in colon cancers.

Results: Sixty nine clones were selected by SSH and macro-DNA array analyses. These clones were then analysed by ISH to examine their expression patterns. ISH screening revealed that all the clones screened showed more intense signals in colon cancers than in non-cancerous tissues. Among them, RACK 1, which is a protein kinase C receptor and a homologue of the G protein ß subunit, was expressed intensely in colon cancer cells. RACK 1 expression was evaluated in multiple samples by ISH, and the results confirmed that RACK 1 was universally overexpressed in cells of all 11 colon cancers examined.

Conclusions: Many genes, including RACK 1, expressed in colon cancer cells can be isolated efficiently by this method, and their precise expression pattern can be evaluated. These results indicate that ISH is an excellent technique for systemic screening of genes expressed in a region specific manner.

Abbreviations: ISH, in situ hybridisation; NCA, non-specific crossreacting antigen; PCR, polymerase chain reaction; RT, reverse transcription; SDS, sodium dodecyl sulphate; SSC, saline sodium citrate; SSH, suppression subtractive hybridisation

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