去甲硫氨酸环境加化疗药物对人胃癌原代细胞生长的阻抑

The experimental studies on the effect of methionine deprivation with chemotherapy to human primary gastric cancer cells

目的 :探索人胃癌原代细胞在体外去除甲硫氨酸 (Met)但含同型半胱氨酸 (Hcy)的培养基 (Met-Hcy+)中能否正常生长和去Met状态是否增加化疗药物对胃癌细胞的杀伤作用。 方法 :将 40例新鲜人胃癌组织制成单细胞悬液 ,分别置于Met-Hcy+和Met+Hcy-(含Met不含Hcy)的培养基中培养 48h ,利用MTT等方法检测各组细胞增殖情况以及在不同培养基中分别加入ADM、DDP、5 FU、MMC和MTX化疗药物后对各组细胞增殖的抑制程度。 结果 :①人胃癌原代细胞在Met-Hcy+培养基中表现为细胞总数减少 ;②Met-Hcy+培养基分别与ADM、DDP、5 FU、MMC和MTX联合时 ,可显著提高各化疗药物对胃癌原代细胞的杀伤作用。 结论 :①人胃癌原代细胞在体外培养中表现出对Met的依赖性 ;②Met-Hcy+培养环境联合应用各化疗药物 ,可提高对人胃癌原代细胞的杀伤作用 ;③MTT法是检测“Met饥饿法”联合个体化化疗药物敏感性的有效手段。

Objectives:To study the methionine dependence(Met dependence) of human primary gastric cancer cells in vitro when Met in the culture medium was replaced by its precursor homocysteine(Hcy),and the effect of methionine starvation in combination with chemotherapeutical agents on gastric cancer cells. Methods:Fresh and sterile gastric cancer samples were managed to single cell suspensions and then were cultured in Met -Hcy + and Met +Hcy - medium separately,the proliferation of tumor cells in different culture media was examined by microcytotoxicity(MTT) assay.Meanwhile,the inhibition rate of tumor cells by ADM、DDP、5 FU、MMC and MTX in Met -Hcy +medium was separately tested. Results:①In Met -Hcy + medium,the human primary gastric cancer cell decreased;②Methionine deprivation in combination with chemotherapy enhanced obviously the killing capacity of each chemotherapeutical agent. Conclusions:①Human primary gastric cancer cells in vitro appears Met dependent.②The combined application of Met -Hcy + medium and different chemotherapeutical agents could enhance the antitumor effect of chemotherapy on primary gastric cancer cells.③MTT assay was an efficient way to examine the sensititivity of methionine starvation therapy combined with individualized chemotherapeutical agents.