Then analysis of lung cancer protein using two-dimensional polyacrylamide gel electrophoresis Application for adjuvant chemotherapy

1999 Fiscal Year Final Research Report Summary

• Project/Area Number: 10671271
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Thoracic surgery
• Research Institution: Tokyo Medical University
• Principal Investigator: HIRANO Takashi Tokyo Medical University, Department of Surgery, Assistant Professor, 医学部, 講師 (30238381)
• Co-Investigator(Kenkyū-buntansha): KONAKA Chimori Tokyo Medical University, Department of Surgery, Assistant Professor, 医学部, 助教授 (70147180) ; KAWATE Norihiko Tokyo Medical University, Department of Surgery, Assistant Professor, 医学部, 講師 (70204740) ; IKEDA Norihiko Tokyo Medical University, Department of Surgery, Assistant Professor, 医学部, 助手 (70246205) ; KATO Harubumi Tokyo Medical University, Department of Surgery, Professor and Chairman, 医学部, 教授 (20074768)
• Project Period (FY): 1998 – 1999
• Keywords: lung cancer protein / antibody / prognosis / TA02 / NapsinA / reticulocalbin / GST-π / Two-dimensional polyacrylamide gel electrophoresis

Research Abstract

1. Sixteen polypeptides associated with histopathological differentiation were detected using two-dimensional polyacrylamide gel electrophoresis of primary lung carcinoma.
2. Based on the expression of the polypeptides associated with either histopathological differentiation or cell proliferation, we classified primar lung adenocarcinoma into typical case and atypical case. 65.4% of the typical cases were diagnosed as well differentiated adenocarcinoma histopathologically. Also, 65.4% of the typical cases showed negative in lymph node metastasis. On the other hand, 20.5% and 34.9% of the atypical cases were judged as well differentiated case and cases without lymph node metastasis, respectively. Furthermore, we recognized the statistically significant difference in three year-disease free survival(62.9% in typical cases, 27.1% in atypical cases ; p=0.0011).
3. We produced monoclonal antibodies against N-terminal of TA02 molecule. The histological distribution of TA02 was clarified immuno histochemically using this antibody and human tissues. Also, we obtained the evidence that TA02 molecule was homologos with NapsinA which was a novel aspartic proteinase published at the end of 1998, because the adenocarcinoma cases with high expression of TA02 showed the expression of napsin A in RNA level. Furthermore, GST fusion protein of Napsin A showed positive reaction with anti-TA02 antibody.
4. TS02 associated with neuroendocrine cells was homologous with protein gene product 9.5(PGP 9.5). In the TS02 positive cases two-years prognosis become worse compared to TS02 negative cases.
5. Compred to the 2DE patterns between CDDP-resistant cell line and the parents cell line, the positions of reticulocarbin and glutathione-S-transferase π were identified on 2DE gel. The expession level of reticulocalbin, which was Ca-binding protein at endoplasmic reticulum, became low in the CDDP-resistant cell line. On the other hand, the expression level of GST-πbecame high, remarkably.
6. The analysis based on lung cancer protein using 2DE is valuable for the evaluation of biological tumor malignancy.