下调上皮内膜蛋白1抑制胶质母细胞瘤恶性生物学行为的实验研究

Experimental research on inhibiting malignant biological behavior of glioblastoma by down-regulating epithelial membrane protein-1

目的研究下调上皮内膜蛋白1(EMP1)对胶质母细胞瘤U251与U87细胞系增殖、侵袭、迁移、凋亡及U87细胞系荷瘤小鼠肿瘤体积及免疫组化指标的影响。方法采用siRNA干扰技术下调胶质母细胞瘤细胞系U87与U251中EMP1的表达。RT-qPCR和Western blot检测EMP1mRNA和蛋白的表达;CCK8和Transwell实验检测EMP1下调后肿瘤细胞增殖迁移及侵袭能力;流式细胞术检测EMP1下调后肿瘤细胞凋亡表达水平;构建U87细胞系裸鼠荷瘤模型,免疫组化法检测Ki-67和MMP2的表达。结果RT-qPCR和Western blot结果显示siRNA-EMP1导致EMP1的表达降低(P<0.05),CCK8和Transewll实验显示下调EMP1后肿瘤细胞的增殖、迁移及侵袭能力被显著抑制(P<0.05);流式细胞术结果显示下调EMP1后肿瘤细胞的凋亡能力明显增强(P<0.05);U87细胞系裸鼠荷瘤模型中下调EMP1后肿瘤细胞生长延缓,Ki-67及MMP2的表达降低。结论EMP1的表达下调可以抑制胶质母细胞瘤的增殖、迁移及侵袭,促进凋亡,延缓荷瘤小鼠肿瘤的生长与侵袭,从而抑制胶质母细胞瘤的恶性生物学行为。

Objective To investigate the impacts of down-regulating epithelial membrane protein-1(EMP1)on proliferation,invasion,migration and apoptosis of U251 and U87 cell lines,and tumor volume and immunohistochemical indices of U87 cell line tumor-bearing mice.Methods The expression of EMP1 in glioblastoma cell lines U87 and U251 was down-regulated by siRNA interference technique.RT-qPCR and Western blot were used to detect the expression of EMP1 mRNA and protein.CCK8 and Transwell experiments were used to detect the proliferation,migration and invasion ability of tumor cells after EMP1 was down-regulated.Flow cytometry was used to detect the apoptosis expression level of tumor cells after EMP1 down-regulation;A nude mouse model of U87 cell line was established,and the expression of Ki-67 and MMP2 was detected by immunohistochemistry.Results RT-qPCR and Western blot results showed that siRNA-EMP1 reduced the expression of EMP1(P<0.05),and CCK8 and Transwell experiments showed that the proliferation,migration and invasion of tumor cells were significantly inhibited after down-regulating EMP1(P<0.05).The results of flow cytometry showed that the apoptosis ability of tumor cells was significantly enhanced after downregulation of EMP1(P<0.05).Downregulation of EMP1 in the U87 cell line nude mouse tumor-bearing model resulted in delayed tumor cell growth.The expression of Ki-67 and MMP2 was reduced.Conclusion Down-regulation of EMP1 expression can inhibit the proliferation,migration and invasion of glioblastoma,promote apoptosis,delay the growth and invasion of tumor-bearing mice,and thus inhibit the malignant biological behavior of glioblastoma.