AKT在Ghrelin抑制结肠癌细胞凋亡中作用.docVIP

AKT在Ghrelin抑制结肠癌细胞凋亡中作用 　　摘要：目的观察生长激素释放肽(growth hormone releasing peptide, Ghrelin)对人结肠癌RKO细胞凋亡的影响及可能机制。方法5-氟尿嘧啶（5-FU）联合Ghrelin作用人结肠癌RKO细胞及预处理AKT抑制剂wortmannin后再作用RKO细胞，Cell Counting Kit-8(CCK-8)法检测细胞增殖；荧光显微镜下观察细胞凋亡；Western blotting检测结肠癌细胞中p-AKT蛋白表达。结果Ghrelin可削弱5-FU对RKO细胞的增殖抑制和凋亡诱导作用，而渥曼青霉素（wortmannin）可显著抑制Ghrelin对体外结肠癌RKO细胞的保护作用；Ghrelin可促进p-AKT蛋白在RKO细胞中的表达，该作用可被wortmannin阻断。结论Ghrelin可显著抑制体外结肠癌RKO细胞凋亡，该作用可能通过促进AKT磷酸化的表达而实现，Ghrelin可能作为结肠癌治疗中新的抗癌靶点。 　　关键词：结肠癌；Ghrelin；AKT 　　Ghrelin Inhibit Cell Apoptosis in Human Colorectal Cancer Cell Line RKO Via AKT Pathway 　　TANG Zhen, LIU An, TANG Min 　　(Institute of Orthpaedics, Yueyang Hospital, The Medical College of Xiangya, The Central South University, Yueyang 414000,Hunan,China) 　　Abstract：ObjectiveTo investigate the anti-apoptotic effect and the mechanism of Ghrelin in human colorectal cancer cell line RKO in vitro. MethodsAfter human colorectal cancer cell line RKO was 5-FU treatment to induce apoptosis and simultaneously treated with Ghrelin in the presence or absence of wortmannin, a highly specific inhibitor of AKT. The cellular proliferation was detected by Cell Counting Kit-8 (CCK-8) assay. The morphological changes of RKO cells were observed under the fluorescence microscopy after DAPI staining. Western blotting was used to detect the expression of p-AKT in RKO cells. ResultsGhrelin prevented apoptosis in RKO cells induced by 5-FU. However, wortmannin abolished Ghrelin s cytoprotective activity after 5-FU treatment. Ghrelin treatment markedly increased AKT activation in RKO cells, and this was completely suppressed by coincubation with wortmannin. ConclusionGhrelin may act as a survival factor in RKO cells through the activation of AKT, and Ghrelin might become a potential therapeutic target of colorectal cancer. 　　Key words： Colorectal cancer; Ghrelin; AKT 　　Ghrelin作为一种生长激素促泌剂受体的内源性配体，其大部分由胃肠道上皮细胞分泌，自从Ghrelin被发现以来，就被证实在包括控制进食和新陈代谢等多种生理及病理活动中也发挥了关键作用。近来来研究表明，Ghrelin在前列腺癌[1]、胃癌[2]和肝癌[3]等多种恶性肿瘤细胞中表达升高，同时对乳腺癌及前列腺癌细胞凋亡有