乙酰辅酶A合成酶2低表达人胃癌细胞株MGC80-3的顺铂敏感性观察  

Cisplatin sensitivity of human gastric carcinoma with low expression of ACSS2

作　　者：糜磊 武丹 陶清 周月鹏[1] 陈德玉[1] LEI Mi;DAN Wu;QING Tao;ZHOU Yuepeng;CHEN Deyu(The Affiliated Hospital of Jiangsu University,Zhenjiang 212001,China)

机构地区：[1]江苏大学附属医院

出　　处：《山东医药》2019年第18期32-35,共4页Shandong Medical Journal

基　　金：江苏省医学创新团队项目(CXTDC2016009);江苏省科技项目(BE2017696)

摘　　要：目的观察乙酰辅酶A合成酶2(ACSS2)低表达人胃癌细胞株MGC80-3的顺铂敏感性,并进一步探讨其可能作用机制。方法 采用Western blotting法检测人胃癌细胞株MGC80-3、AGS、SGC-7901、NCI-N87以及正常胃黏膜上皮细胞系RGM-1中ACSS2蛋白。对数生长期MGC80-3细胞分为A、B、C、D组,B及D组分别加入10 μL脂质体Lipofectamine 2000+5 μL siRNA-ACSS2转染, C组加入及A组分别加入50 μL无血清纯RPMI1640培养基+5 μL 阴性对照(NC);A组、B组均培养72 h,C组培养48 h时加入5 μg/mL的顺铂(DDP),D组转染48 h时加入5 μg/mL的DDP。培养24 h时,CCK8法检测各组细胞增殖活力,培养48、72 h时流式细胞分析术测算各组细胞早晚期凋亡率,Western blotting法检测各组细胞ACSS2、p65、p-p65、B淋巴细胞瘤-2蛋白(Bcl-2)、B细胞淋巴瘤-特大型蛋白(Bcl-xL)、凋亡蛋白Bcl-2相关的X蛋白(Bax)。结果 MGC80-3、AGS、SGC-7901、NCI-N87及RGM-1细胞ACSS2蛋白相对表达量分别为0.80± 0.03 、0.62±0.04、0.60±0.04、0.72±0.05、0.41±0.04。与RGM-1细胞比较,MGC80-3、AGS、SGC-7901、NCI-N87细胞ACSS2相对表达量升高( P 均<0.01)。A、B、C、D组细胞增殖活力分别为98.67%±4.89%、92.67%±7.86%、62.67%± 4.05%、34.67%±7.61%,A组与B组相比无明显差异( P >0.05);与A组比较,C组细胞增殖活力下降( P < 0.001 );与B、C组比较,D组细胞增殖活力均下降( P 均< 0.001 )。A、B、C、D组早期细胞凋亡率分别为1.06%±0.02%、1.13%±0.08%、7.62%±0.55%、14.33%±0.21%,晚期细胞凋亡率分别为8.01%± 0.05%、8.13%±0.06%、10.81%±0.43%、15.98±0.18%。与A组比较,C、D组细胞早、晚期凋亡率均升高( P 均<0.05);与B组比较,D组早、晚期凋亡率均升高( P 均<0.05);与C组比较,D组细胞早、晚期凋亡率均升高( P 均< 0.05 )。与A组比较,C组p-p65、Bax蛋白相对表达量升高,Bcl-2、Bcl-xL相对表达量降低( P 均<0.05);与B组比较,D组细胞p65、p-p65、Bcl-2、Bcl-xL相对表达量均降低、Bax相�Objective To observe the cisplatin sensitivity of human gastric cancer cell lines with low expression of acetyl-CoA synthetase 2 (ACSS2) and to explore its possible mechanism. Methods The expression of ACSS2 in the gastric cancer cells MGC80-3, AGS, SGC-7901, NCI-N87, and the normal gastric mucosal epithelial cells RGM-1 was detected by Western blotting. MGC80-3 cells in the logarithmic growth phase were divided into groups A, B, C, and D;cells in groups B and D were added with 10 μL Lipofectamine 2000+5 μL siRNA-ACSS2 for transfection;cells in groups C and A were added with 50 μL serum-free pure RPMI1640 medium + 5 μL negative control (NC);cells in the group A and group C were cultured for 72 h;5 μg/mL cisplatin (DDP) was added into the group C after 48 h, and 5 μg/mL DDP was added into the group D after 48 h. When cultured for 24 h, the proliferation of each group was detected by CCK8. The early and late apoptosis rates of cells was measured by flow cytometry at 48 and 72 h. Western blotting was used to detect the expression levels of ACSS2, p65/p-p65, B lymphocyte tumor-2 protein (Bcl-2), B-cell lymphoma-extra large protein (Bcl-xL), and apoptosis protein Bcl-2 related X protein (Bax). Results The relative expression levels of ACSS2 in the MGC80-3, AGS, SGC-7901, NCI-N87 and RGM-1 cells were 0.80±0.03, 0.62±0.04, 0.60±0.04, 0.72 ±0.05, and 0.41± 0.04 , respectively. The relative expression of ACCS2 in the MGC80-3, AGS, SGC-7901 and NCI-N87 cells increased as compared with that of the RGM-1 cells (all P <0.01);The cell proliferation viability of the groups A, B, C, and D were 98.67%±4.89%, 92.67%±7.86%, 62.67%±4.05%, and 34.67%±7.61%, respectively. There was no significant difference between the groups A and B ( P >0.05). Compared with the group A, the cell proliferation of group C decreased ( P <0.001). Compared with the groups B and, the cell proliferation of group D decreased ( P <0.001). The early apoptosis rates of the groups A, B, C, and D were 1.06%±0.02%, 1.13%±0.08%, 7.62%± 0.55%, and

关 键 词：乙酰辅酶A合成酶2 核因子κB B淋巴细胞瘤-2蛋白 B细胞淋巴瘤-特大型蛋白 Bcl-2相关的X蛋白 胃癌 顺铂 化疗敏感性 

分 类 号：R735.2[医药卫生—肿瘤]