Development of a novel strategy for mutation-specific cloning using DNA repair enzyme

• Project/Area Number: 16510149
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied genomics
• Research Institution: Kitasato University
• Principal Investigator: TSUKAMOTO Toshihiko Kitasato University, School of Pharmaceutical Sciences, Associate Professor, 薬学部, 助教授 (10236862)
• Project Period (FY): 2004 – 2005
• Project Status: Completed (Fiscal Year 2005)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,900,000); Fiscal Year 2005: ¥1,100,000 (Direct Cost: ¥1,100,000); Fiscal Year 2004: ¥2,800,000 (Direct Cost: ¥2,800,000)
• Keywords: Mutation / クローニング / 大腸癌

Research Abstract

In this study, we pulled down heteroduplexes formed between cDNA libraries constructed from a colorectal cancer with high microsatellie instability (MSI-H) and paired normal mucosa using a recombinant MutS protein and isolated paires of mutant and wild type sequences involving ; CEA (carcinoembryonic antigen-related cell adhesion molecule 5 ; Hs.220529) 3'UTR, 2780 A to G ; 2792-2798 (T) 7 to (T) 8 (GI : 178676 ; CDS 97..2205) ; CCNI (Cyclin I ; Hs.486360) 3' UTR, 1800 (A) 10 to (A) 9 (GI : 13436391 ; CDS 544..1677) ; PTMA (prothymosin, alpha ; Hs.264317) 5'UTR, 142 T to C ; CDS, 228 T to C (GI : 47938172 ; CDS 161..493) ; SRI (sorcin ; Hs.489040) CDS 488 A->G, Y159C (GI : 4507206 ; CDS 13..609) ; LGALS3 (lectin, galactoside-binding, soluble, 3 ; galectin 3 ; Hs.531081) 774 A to G, 1250M (GI : 37589086 ; CDS 25..777) ; RACK1 (GNB2L1, guanine nucleotide binding protein (G protein), beta polypeptide 2-like 1 ; Hs.5662) 288t->g, F65V, 304t->c, V70A (GI : 187701 ; CDS 96..1049) ; UQCRQ (ubiquinol-cytochrome c reductase, complex III subunit VII, 9.5kDa ; Hs.146602) CDS 227 t->C, F67S, (GI:34784784 ; CDS 28..276) ; and also isolated a novel NAT highly mutated in MSI-H tumors for thymidylate synthase.

Research Products

• [Journal Article] cDNA array analysis for diagnosis of hepatic metastasis of colorectal carcinoma using tissue samples obtained from primary tumors.
  • Author(s): Takata, O., Kawamura, Y., Konishi, F., Sasaki, J., Kai, T., Miyakura, Y., Nagai, H., Tsukamoto, T.
  • Journal Title: Surgery Today In press
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Overexpression in colorectal cancer of two lysosomal enzymes, CLN2 and CLN1, involved in neuronal ceroid lipofuscinosis.
  • Author(s): Tsukamoto, T., Iida, J., Dobashi, Y., Furukawa, T., Konishi, F.
  • Journal Title: Cancer In press
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Clinicopathological characteristics of colorectal cancers with loss of imprinting of insulin-like growth factor 2.
  • Author(s): Sasaki, J., Konishi, F., Kawamura, Y, Kai., T., Takata, O., Tsukamoto, T.
  • Journal Title: Int J Cancer In press
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] The majority of human aberrant crypt foci in the lower rectum are precursors of hyperplastic polyps.
  • Author(s): Togashi, K., Konishi, F., Kojima, M., Nagai, H., Tsukamoto, T.
  • Journal Title: Dis Colon Rec In press
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] cDNA array analysis for diagnosis of hepatic metastasis of colorectal carcinoma using tissue samples obtained from primary tumors.
  • Author(s): Takata, O., Kawamura, Y., Konishi, F., Sasaki, J., Kai, T., Miyakura, Y., Nagai, H., Tsukamoto, T.
  • Journal Title: Surgery Today (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Overexpression in colorectal cancer of two lysosomal enzymes, CLN2 and CLN1, involved in neuronal ceroid lipofuscinosis.
  • Author(s): Tsukamoto, T., Iida, J., Dobashi, Y., Furukawa, T., Konishi, F.
  • Journal Title: Cancer (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Clinicopathological characteristics of colorectal cancers with loss of imprinting of insulin-like growth factor 2.
  • Author(s): Sasaki, J., Konishi, F., Kawamura, Y, Kai., T., Takata, O., Tsukamoto, T.
  • Journal Title: Int J Cancer. (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The majority of human aberrant crypt foci in the lower rectum are precursors of hyperplastic polyps.
  • Author(s): Togashi, K., Konishi, F., Kojima, M., Nagai, H., Tsukamoto, T.
  • Journal Title: Dis Colon Rectum (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] cDNA array analysis for diagnosis of hepatic metastasis of colorectal carcinoma using tissue samples obtained from primary tumors.
  • Author(s): Takata, O., Kawamura, Y., Konishi, F., Sasaki, J., Kai, T., Miyakura, Y., Nagai, H., Tsukamoto, T.
  • Journal Title: Surgery Today (In press)
• [Journal Article] Overexpression in colorectal cancer of two lysosomal enzymes, CLN2 and CLN1, involved in neuronal ceroid lipofuscinosis.
  • Author(s): Tsukamoto, T., Iida, J., Dobashi, Y., Furukawa, T., Konishi, F.
  • Journal Title: Cancer (In press)
• [Journal Article] Clinicopathological characteristics of colorectal cancers with loss of imprinting of insulin-like growth factor 2.
  • Author(s): Sasaki, J., Konishi, F., Kawamura, Y., Kai, T., Takata, O., Tsukamoto, T.
  • Journal Title: Int J Cancer (In press)
• [Journal Article] The majority of human aberrant crypt foci in the lower rectum are precursors of hyperplastic polyps.
  • Author(s): Togashi, K., Konishi, F., Kojima, M., Nagai, H., Tsukamoto, T.
  • Journal Title: Dis Colon Rec (In press)