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 anti-human 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 anti-body could effectively inhibit the proliferation of HNE-1 cells,promote apoptosis,and arrest cell cycle.Anti-human 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.%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.
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