谷氨酰胺酶的活性检测和肿瘤个性化治疗的探索

来源 :2015中国转化医学大会暨中国转化医学联盟成立大会 | 被引量 : 0次 | 上传用户:Vince6666
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  肿瘤细胞基因突变,特别是糖酵解中的丙酮酸激酶肌肉同工酶剪切变构体2(PKM2)突变,可导致葡萄糖有氧糖代谢途径的明显改变,即Warburg效应:葡萄糖代谢增加200倍,但产物乳酸被排出体外,不能进入线粒体三羧酸循环,则谷氨酰胺成为肿瘤细胞线粒体产生能量所依赖的原料,而谷氨酰胺酶是谷氨酰胺进入线粒体三羧酸循环必不可少的酶.谷氨酰胺缺乏或谷氨酰胺酶的抑制均能抑制肿瘤细胞的生长.,但肿瘤细胞的差异性使其对谷氨酰胺酶抑制剂的敏感性亦不相同,因此对谷氨酰胺酶抑制剂的肿瘤治疗进行个性化治疗研究是非常有必要的.限于目前谷氨酰胺或谷氨酰胺酶的定量方法仅有酰肼340nm波长吸光度检测法、谷氨酸在氧化酶氧化荧光波长检测法、固定化谷氨酰胺酶膜氨水含量检测法和放射性物质标记法,而这些方法均有灵敏度不高、干扰大等局限因素.我们合成了多种异硫氰酸荧光素标记的荧光化合物,建立一种新颖快速的定量检测方法,并将其应用于探索肿瘤细胞中谷氨酰胺线粒体代谢与谷氨酰胺酶抑制剂敏感性的关系.为肿瘤精准的个性化治疗提供了一种干扰少、快速、准确、灵敏的分析方法.
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