Comparison Of TK Gene Mutation Induced By Vinblastin And Colcemid And Study On The Mechanism For Two Kinds Of Mutants

Since aneuploidy is a serious genetic problem in human health, detection of potential chemicals to induce aneuploidy is very important for human health.As so far there is no related reports about assessment genetoxicity of spindle poisons with TK gene mutation assay using human lymphoblastoid cells. The object of the research is to study the detection ability of aneuploidy with TK gene mutation assay using human lymphoblastoid cellsTK6 and TK6-E6, and to assess the affection of P53 protein on the assay system, to explore the mechanism resulting in the formations of normal growing colony and slowly growing colony by cytological and molecular-biological methods.This study includes three parts.Part 1.Comparison of TK Gene Mutation Assay Induced by Vinblastin with 3h treatment and 24h treatment. The object of the part is to compare TK gene mutation assay induced by vinblastin with 3h treatment and 24h treatment in human lymphoblastoid cells TK6. The results of the study showed that the mutegenecity of vinblastin in TK6 cells was negative with 3h treatment, while positive with 24h treatment, which indicated that longer exposure may be needed for spindle poisons and other cell cycle dependent chemicals.Part 2. Comparison of TK Gene Mutation Assay in TK6 and TK6-E6 Cell Induced by Two Spindle Poison. The object of this study is to compare the detection ability of aneuploidy with TK gene mutation assay between human lymphoblastoid cells TK6 and TK6-E6 and the affection of P53 protein for the detection system. The results showed that the cytotoxicity RS, RV, RSG and RTG decreased with the concentration of vinblastin and colcemid increasing, indicating that TK6 cells was more sensitive to vinblastin and colcemid than TK6-E6 cells. Except TK6-E6 group treated with colcemid, mutation frequency increased significantly in TK6 and TK6-E6 cell induced by Vinblastin and Colcemid, and except CCM5.0 ng/ml group, the mutation frequency of TK6 were higher than that of TK6-E6 at same dose (p<0.05), indicating that Both TK6 and TK6-E6 cell lines can be used in TK gene mutation assay to access the spindle poison. The RS, RSG, RTG and MF of TK6-E6 were higher than that of TK6, and TK6-E6 cell line was recommended to be used in the assay for its relative shorter doubling time and less plating cell number.Part 3 Study on the Mechanism for Normal Growing Colony and Slowly...