摘要
目的:探讨S100结合域的缺失对钙周期素结合蛋白(CacyBP/SIP)生物学功能的影响。方法:构建野生型CacyBP/SIP及S100结合域截短突变的真核表达载体(分别为pFLAG-CacyBP和pFLAG-△S100),脂质体法转染至人胃癌细胞系MKN45,四唑盐(MTT)比色法观察细胞生长,平板克隆实验及裸鼠成瘤实验检测转染细胞的体外和体内成瘤性。结果:成功构建了野生型CacyBP/SIP及△S100截短突变的真核表达载体并转染MKN45细胞。野生型CacyBP/SIP及△S100截短突变的CacyBP/SIP均能抑制胃癌细胞MKN45的增殖,使MKN45细胞的体外及体内成瘤性降低,但是截短突变CacyBP/SIP抑制胃癌细胞增殖及成瘤性的作用更强。结论:CacyBP/SIP能够抑制胃癌细胞的恶性表型,而阻断S100分子与CacyBP/SIP的结合能增强CacyBP/SIP这种抑制作用。
OBJECTIVE:To explore the effect of S100 binding domain on the biological behavior of CacyBP/SIP.METHODS:The eukaryotic expression vectors of the wild-type CacyBP/SIP and truncated mutant lacking S100 binding domain required for association with S100 proteins were constructed and transfected into gastric cancer cell line MKN45 by using liposome.The cell growth was determined by MTT,and plate clonogenic assay and tumor xenograft in nude mice were performed respectively to test tumorigenecity of the cells in v...
出处
《中华肿瘤防治杂志》
CAS
2008年第13期968-971,共4页
Chinese Journal of Cancer Prevention and Treatment
基金
国家自然科学基金(30572113)
西京医院科技创新基金(XJCX05M21)