目的 观察serglycin稳定干扰后对鼻咽癌高转移细胞顺铂敏感性的影响,并初步探究其机制。方法 建立serglycin稳定干扰的鼻咽癌高转移细胞株,qRT-PCR 和 Western blot检测serglycin的表达情况。四甲基偶氮唑蓝法测定顺铂对稳定株细胞的IC50;增殖曲线实验观察在顺铂压力下细胞增殖的差异;克隆形成实验观察细胞克隆形成能力的差异; Annexin V/PI 双染检测细胞凋亡。qRT-PCR检测稳定株细胞中干性基因的表达情况。结果 Serglycin稳定干扰的鼻咽癌高转移细胞株成功建立,细胞形态发生改变。四甲基偶氮唑蓝法测得serglycin稳定干扰细胞的顺铂IC50显著低于对照组细胞,增殖抑制率高于对照组,药物作用下克隆形成能力减弱,流式凋亡比例增高。Serglycin稳定干扰后干性基因表达下调。结论 Serglycin稳定干扰可以增加鼻咽癌高转移细胞对顺铂的敏感性。
Abstract
OBJECTIVE To investigate the effect of serglycin knockdown on the sensitivity to cisplatin in nasopharyngeal carcinoma highly metastatic cells. METHODS Stable transfected nasopharyngeal carcinoma highly metastatic cell lines was established. The alteration of serglycin expression was examined by qRT-PCR and Western blot assay established. The IC50 of cisplatin in stable cells were measured by MTT assay. The effect of serglycin knockdown on the proliferation of stable cells treated with cisplatin were measured by MTT assay. Colony formation assay were used to show cloning capacity. Annexin V/PI staining were used to detect apoptosis. qRT-PCR analysis was performed for stem cell-associated genes in stable cells. RESULTS Stable transfected cell lines were successfully established and cellular morphology of stable transfected cells were changed. When treated with cisplatin, the IC50 and the colonies of serglycin knockdown cells were significant lower than controlled cells, however, the inhibition of proliferation and the percentage of apoptosis were significant higher than controlled cells. The stem cell-associated genes were reduced in serglycin knockdown cells. CONCLUSION Stable interference of serglycin makes nasopharyngeal carcinoma highly metastatic cells more sensitive to cisplatin.
关键词
serglycin /
鼻咽癌 /
顺铂
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Key words
serglycin /
nasopharyngeal carcinoma /
cisplatin
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中图分类号：
R965
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参考文献
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脚注
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基金
广东省科技计划项目(2012B020306001)
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