Gene Therapy of p53-inactivating tumor by Caspase3

2003 Fiscal Year Final Research Report Summary

• Project/Area Number: 14571217
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Digestive surgery
• Research Institution: Nagoya City University
• Principal Investigator: SHINODA Noriyuki Nagoya City University, Graduate School of Medical Sciences, Research Associate, 大学院・医学研究科, 助手 (80305549)
• Co-Investigator(Kenkyū-buntansha): FUJII Yoshitaka Nagoya City University, Graduate School of Medical Sciences, Professor and Chair, 大学院・医学研究科, 教授 (40156831) ; KUWABARA Yoshiyuki Nagoya City University, Graduate School of Medical Sciences, Assistant Professor, 大学院・医学研究科, 講師 (90225326)
• Project Period (FY): 2002 – 2003
• Keywords: Caspase3 / apoptosis / XIAP / anti-cancer therapy / VP16

Research Abstract

Only in the neoplasm cell which p53 was mutated and inactivated, this research devised the system by which Caspase3 is activated, and aimed at improvement in the anti-cancer effect of neoplasm. First, we carried out the cloning of the expression vector of CaspaseS. Moreover, on the cancer cell lines which has p53 mutation, we checked that Caspase3 by this vector was not expressed. Moreover, we checked that included XIAP gene in the down-stream of biding site of p53,expression of XIAP went up on a normal cell line, and expression did not go up on the colon cancer cell stock SW480 with mutant type p53. Next, we checked whether a XIAP vector and a Caspase3 expression vector would be simultaneously introduced into a cancer cell line (wild type p53), and a normal cell line under VP16 existence using these vectors, and apoptosis could be avoided. The analysis of apoptosis was carried out by the increase in subG1 population using FACS. When the XIAP vector and Caspase3 vector were co-transfec ted into the human skin normal cell line NHDF, apoptosis was not induced to NHDF. However, when both vectors were transfected into the colon cancer cell line SW480 which has the mutant type p53 under VP16 existence, apoptosis was not induced for four days. Moreover, apoptosis was not induced although transfection was tried to other cell lines, DLD1,HCT116,etc. Although we thought these results that each vector was expressed well, apoptosis has not been induced to cancer-and normal tissues selection system only with these two genes. Although the cause cannot be solved, the relation between that intervention of another factor(s) besides XIAP is mentioned as a candidate and control of XIAP of Caspase3 may have regulation of the delicate amount of expression. In this system, apoptosis was not able to be induced only into the cell which has mutation in p53. If the mechanism of the apoptosis by Caspase3 will become still clearer from now on, we want to carry out cloning to the vector which can fine-adjust change or the amount of expression, or change the vector XIAP to another gene using the same system, and to find the way to the therapy.