抗人IgM抗体对鼻咽癌HNE-1细胞生物学特性的影响

Effect of anti-human IgM antibody on biological characteristics of human nasopharyngeal carcinoma HNE-1 cell line in vitro and in vivo

目的研究抗人免疫球蛋白M(IgM)抗体对鼻咽癌HNE-1细胞增殖、凋亡、细胞周期和成瘤的影响。方法经抗人IgM抗体处理后,采用细胞增殖抑制实验观察HNE-1细胞增殖,流式细胞术检测细胞凋亡及周期,脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(TUNEL)染色检测细胞凋亡;构建裸鼠动物模型,腹腔注射抗人IgM抗体,观察移植瘤的生长,免疫组织化学SP法检测移植瘤中IgM和糖蛋白96(gp96)的蛋白表达。结果抗人IgM抗体可呈浓度和时间依赖性抑制HNE-1细胞的增殖(P<0.05);流式细胞术检测表明抗人IgM抗体可促进HNE-1细胞G1期细胞百分比明显降低,S期细胞百分比明显增加,细胞凋亡率明显增高(P<0.05);TUNEL检测提示抗人IgM抗体可促进HNE-1细胞凋亡(P<0.01);移植瘤实验显示抗人IgM抗体可明显抑制移植瘤的体积和质量(P<0.05);移植瘤免疫组织化学显示裸鼠腹腔注射抗人IgM抗体后,移植瘤内IgM和gp96蛋白的表达水平明显低于对照组(P<0.05)。结论抗人IgM抗体可有效抑制HNE-1细胞的增殖、促进细胞凋亡、阻滞细胞周期,并且能抑制裸鼠移植瘤的生长,其机制可能与抑制IgM和gp96蛋白表达有关。

Objective To investigate the effect of anti-human immunoglobulin M（IgM）on proliferation,apoptosis,cell cycle and tumor formation in human nasopharyngeal carcinoma HNE-1 cell line in vitro and in vivo.Methods After treatment with anti-human IgM antibody,proliferation of HNE-1 cells was observed by cell proliferation inhibition assay,apoptosis and cell cycle of HNE-1 cells were detected by flow cytometry,and apoptotic cells were detected by TUNEL staining.Nude mouse models were constructed,and were injected intraperitoneally with anti-human IgM antibodies（once every 3 days）.The growth of transplanted tumor was observed once every 4 days.After the fifth injection,the expression levels of IgM and gp96 protein in transplanted tumor were observed by immunohistochemical method（streptavidin-peroxidase conjugated method,SP）.Results MTS assay showed that anti-human IgM antibody can significantly inhibit the proliferation of HNE-1 cells in concentration-and time-dependent manner（P〈0.05）.Flow cytometry showed that the antihuman IgM antibody promoted a significant decrease in percentage of cells in G1 phase,a significant increase in percentage of cells in S phase,and a significant increase in apoptotic rate of HNE-1 cells（P〈0.05）.TUNEL staining showed that the anti-human IgM antibody promoted apoptosis of HNE-1 cells（P〈0.01）.Transplantation tumor experiment showed that anti-human IgM antibody can significantly inhibit the volume and weight of transplanted tumor（P〈0.05）.The immunohistochemistry showed that the expression levels of IgM and gp96 proteins in mouse transplanted tumors after intraperitoneal injection with anti-human IgM antibodies were significantly lower than those of the control group（P〈0.05）.Conclusion The anti-human IgM antibody could effectively inhibit the proliferation of HNE-1 cells,promote apoptosis,and arrest cell cycle.Antihuman IgM antibody could also inhibit the growth of transplanted tumor in nude mouse,which might be related to inhibition of the expressions of IgM and gp96 proteins.