LY294002抑制PI3K/AKT信号通路对胃腺癌SGC-7901细胞的影响

韩静; 张庆瑜; 康春生; 谢甲贝; 付彦超; 张靖; 马菲菲; 王涛

doi:10.3969/j.issn.1000-8179.2011.05.004

摘要: 目的：观察应用PI3K抑制剂2- （4-吗啉基） -8-苯基-4氢-1-苯并吡喃-4酮（LY294002）对胃腺癌细胞株SGC-7901中磷脂酰肌酸3-激酶（PI3K） /丝氨酸苏氨酸激酶（AKT）信号通路的变化以及由此产生的细胞生长、侵袭及凋亡等方面的影响。方法：应用LY294002作用于胃腺癌细胞株SGC-7901，通过Western blot检测PI3K/AKT信号通路中相关蛋白质P-AKT、基质金属蛋白酶-2 （MMP-2）、基质金属蛋白酶-9 （MMP-9）、 B细胞淋巴瘤/白血病-2 （Bcl-2）、细胞周期素（CyclinD1）的表达情况。噻唑蓝比色法（MTT）和流式细胞法及Transwell等实验评价胃腺癌细胞增殖、侵袭、凋亡等变化。结果：与空白对照组和DMSO组相比，LY294002组能有效抑制P-AKT、 MMP-2、 MMP-9、 Bcl-2及CyclinD1蛋白的表达。LY294002组自第2天起生存率明显下降（P<0.05）。Transwell穿过细胞数结果分别为空白对照组（81.0±2.65）、 DMSO组（81.3±1.52）、 LY294002组（44.6±2.52）， 3组相比较差异具有统计学意义（F=255.1， P=0.000）。LY294002组细胞周期被阻滞在G0/G1期， S期减（P<0.05），早期凋亡的细胞数明显增多（F=149.66， P=0.000）。结论： LY294002是一种对PI3K/AKT信号传导通路具有抑制作用的抑制剂，靶向PI3K的LY294002可以有效抑制胃腺癌SGC-7901细胞的PI3K/AKT信号通路中相关蛋白质的表达，在体外显著抑制细胞的增殖、侵袭，并促进细胞的凋亡，因此为临床肿瘤防治提供了新思路，针对信号传导通路的靶向治疗有望成为肿瘤治疗的新途径。

Abstract: Effect of Inhibitory Action of LY294002 in the PI3K/AKT Signaling Pathway on GastricAdenocarcinoma SGC-7901 CellsJing HAN1, Qingyu ZHANG1, Chunsheng KANG2, Jiabei XIE1, Yanchao FU1, Jing ZHANG1, Feifei MA2, TaoWANG1Correspondence to: Qingyu ZHANG, E-mail: zhangqy@tijmu.edu.cn; Chunsheng KANG, email: kangchunsheng@sohu.com1Department of Gastroenterology, Tianjin Medical University General Hospital, Tianjin 300052, China2Department of Neurosurgery and Tumor Section of Neurology Institute, Tianjin Medical University General Hospital, Tianjin 300052, China3Tianjin Medical College, Tianjin 300222, ChinaAbstract Objective: To observe the effects of phosphatidyl creatine 3-kinase (PI3K) inhibitor LY294002 on the PI3K/SerineThreonine Kinases ( AKT ) signal pathway, cell growth, cell invasion and apoptosis in human gastric adenocarcinoma SGC-7901 cells.Methods: Human gastric carcinoma SGC-7901 cells were treated with LY294002, and Western blot was used to detect the expression ofproteins in the PI3K pathway including P-AKT, matrix metalloproteinase-2 ( MMP-2 ), matrix metalloproteinase-9 ( MMP-9 ), B-celllymphoma/leukemia ( Bcl-2 ) and Cyclin D1. Proliferation, invasion and apoptosis were detected by MTT, Transwell assay and flow cy-tometry. Results: Compared with SGC-7901 cells in the control group and DMSO group, LY294002 significantly downregulated the ex-pression of P-AKT, MMP-2, MMP-9, Bcl-2 and CylinD1. Proliferative activity was suppressed in the cells treated with LY294002 startingon the second day, with an obvious decrease in viability ( P < 0.05 ). The results of the Transwell assay showed that the number of cellstravelling through the matrigel was 81.0±2.65 in the control group, 81.3 ±1.52 in the DMSO group, and 44.6±2.52 in the LY294002 group.There were statistical differences among the three groups ( F = 255.1, P = 0.000 ). The flow cytometry results showed that there was G0/G1arrest in the LY294002 group ( χ2= 18.729, P = 0.016 ). The percentage of apoptotic cells was increased when SGC-7901 cells were treatedwith LY294002 ( F = 149.66, P = 0.000 ). Conclusion: LY294002 inhibits the PI3K/AKT signal transduction pathway. LY294002 caneffectively downregulate downstream protein expression, inhibit proliferation and invasion of SGC-7901 cells and increase apoptosis.Keywords LY294002; SGC-7901 cell; PI3K/AKT signal pathway