摘要
目的 p55γ是磷脂酰肌醇3 激酶(PI3K)的调节亚单位之一,在调节PI3K活性上具有重要的作用,本研究旨在观察p55γN末端24个氨基酸对胃癌细胞增殖的影响。方法 用脂质体介导将表达质粒pEGFPC1和pEGFPC24转染MGC 803细胞,通过荧光显微镜鉴定转染基因的蛋白表达;MTT法观察转染细胞的生长情况;软琼脂集落形成实验确定单个细胞的增殖力;最后用流式细胞仪分析细胞周期时相的变化。结果 (1)建立了稳定表达GFP和GFP N24的MGC 803细胞系,分别命名为C1/803和C24/803。(2)与对照组C1/803相比,C24/803细胞生长速度减慢,其在软琼脂中集落形成率下降了86.8%。(3)C1/803和C24/803细胞周期G0 /G1期百分率分别为52.5%和59.5%,两者相比差异有显著性(P<0.05)。结论 p55γN末端24个氨基酸能够抑制胃癌细胞的生长和增殖,引起细胞周期G1 /S期延长,提示N24p55γ可能成为治疗胃癌的潜在的分子药物。
Objective p55 gamma is one of the regulatory subunits of phosphoinositide-3 kinase(PI3K), which plays an important role in regulation of PI3K activity. This study was designed to elucidate the effects of the N-terminal 24 amino acids of p55 gamma on the growth and proliferation of gastric carcinoma. Methods Using the plasmids of pEGFPC1and pEGFPC24 to transfect MGC-803 cell line by Lipofectamine. The expression of transfected genes were identified by fluorescence microscope; the curve of cells growth was determined by MTT assay; the ability of proliferation of MGC-803 cells was measured by the rate of cloning formation; cell cycle of MGC-803 cells was assayed by flow cytometry. Results Transfected MGC-803 cell lines were established, which can stably expressed GFP and GFP-N24. We designated them as C1/803and C24/803 respectively. ②Compared with the control cells C1/803, the growth rate of C24/803 was decreased. The rate of cloning formation in soft agar of C24/803 descended 86.8%. ③The percentage of C1/803 and C24/803 cells in G 0/G 1 phase were 52.5% and 59.5%, respectively. There was significant difference between them ( P < 0.05). Conclusion The results demonstrated that the N-terminal 24 amino acids of the p55(N24 p55) gamma inhibit the growth and proliferation of gastric carcinoma through arresting the cell cycle at G1/S checkpoint. It suggests that N24p55 gamma may be potential molecular drugs for the treatment of gastric carcinoma.
出处
《肿瘤》
CAS
CSCD
北大核心
2005年第1期24-27,共4页
Tumor
基金
国家自然科学基金资助项目(编号: 30171089)
关键词
胃肿瘤
PI3K
p55γ
转染
MGC-803细胞
细胞周期
细胞增殖
Stomach neoplasms
Phosphoinositide-3 kinase
P55 gamma
Transfection
MGC-803 cells
Cell cycle
Cell proliferation
