热CO_2气腹处理后结肠癌细胞株COLO 205的蛋白组学变化

Proteomics Changes of Colonic Cancer Cell COLO 205 after Treatment with Hyperthermic CO_2 Pneumoperitoneum

目的:研究热CO_2气腹处理后结肠癌细胞株COLO 205的蛋白组学变化,为阐明热CO_2气腹对结肠癌细胞的杀伤作用机制提供依据。方法:用热CO_2气腹处理结肠癌细胞株COLO 205,按有无处理分为处理组与对照组。分别抽提两组细胞的总蛋白质,采用同位素标记的相对和绝对定量(isobaric tags for relative absolute quantitation,iTRAQ)技术标记,液相色谱(LC)分离蛋白质,质谱仪(MS)进行蛋白质鉴定及Western b1ot检测。结果:共筛选得到18个差异表达的蛋白,其中8个蛋白表达上调,10个蛋白表达下调。Western blot显示热休克蛋白HSP70在细胞表达明显高于对照组(P<0.01);蛋白Myosin-9的表达量在处理后显著下降。结论:热CO_2气腹处理后结肠癌细胞株COLO 205的蛋白组学发生差异性变化。

Objective： To study the proteomics changes of colonic cancer cell COLO 205 after treatment with hyperthermic COs pneumoperitoneum, and to define the mechanism of the cell-killing effect of it on the cells. Methods： The total protein of colonic cancer cell COLO 205 was extracted and labeled by isobaric tags for relative and absolute quantitation OTRAQ）. The labeled proteins were isolated by liquid chromatography tandem mass spectrometry（LC-MS）on linear ion trap mass spectrometer in integrated mode comprising pulsed-Q-dissociation （PQD） to identify the differentially expressed proteins. The differentially expressed proteins were analyzed by Western blot. Results： Eighteen proteins were identified,among which 8 were highly expressed and 10 were lowly expressed after treatment by HT-CO2. Western blot showed that expression of HSP70 was increased and expression of Myosin-9 was decreased compared to the control group. Conclusion： The proteome profiles of colonic cancer cell COLO205 before and after treatment with HT-CO2 were differentially changed.