LHPP的缺失促进胶质母细胞瘤细胞的增殖迁移并导致患者预后不良

The deficiency of LHPP promoted the growth and migration of glioblastoma and lead to poor prognosis in patients

目的分析磷酸赖氨酸磷酸组氨酸无机焦磷酸盐磷酸酶(LHPP)在胶质母细胞瘤中的表达以及与患者总体生存率的关系,并探究其对胶质母细胞瘤U87MG细胞增殖、凋亡和迁移能力的影响。方法通过GEIPA、CGGA网站分析LHPP在肿瘤组织的表达情况及对患者总体生存率的影响;选择U87MG细胞,设置阴性对照组(NC组)和实验组(siRNA-LHPP组),通过实时荧光定量PCR及蛋白印迹法验证siRNA(siRNA-LHPP)的干扰效果;采用CCK8法和流式细胞术检测干扰LHPP对U87MG细胞增殖和凋亡的影响;Transwell法比较实验组U87MG细胞迁移能力的变化;蛋白印迹法检测干扰LHPP对U87MG细胞内转化生长因子β/Smad2(TGF-β/Smad2)通路相关蛋白的影响。结果生物信息学结果显示,LHPP在高级别胶质瘤中转录和表达下降,导致患者生存期缩短,差异均有统计学意义(P均<0.001)。与阴性对照组比较,实验组细胞的LHPP mRNA转录和蛋白表达水平下调60%,差异均有统计学意义(t=9.090、9.568,P均<0.001)。与阴性对照组比较,实验组细胞的增殖能力增强约50%,迁移能力增强约100%,Ki-67、TGF-β、磷酸化Smad2(p-Smad2)蛋白表达显著增加,差异均有统计学意义(P均<0.05)。加入氧化苦参碱后,TGF-β、p-Smad2的表达下调,LHPP沉默后引起的细胞增殖和迁移能力的增强被显著逆转。结论LHPP的缺失激活了TGF-β/Smad2信号途径,促进胶质母细胞瘤增殖和转移,导致预后不良。

Objective To analyze the expression of phospholysine phosphohistidine inorganic pyrophosphate phosphatase(LHPP)in glioblastoma and its relationship with overall survival rate of patients,and explore its impact on the proliferation,apoptosis,and migration ability of U87MG glioblastoma cells.Methods The expression of LHPP in tumor tissue and its impact on overall survival rate of patients were analyzed through GEIPA and CGGA websites.U87MG cells were used to set up a negative control group(NC group)and an experimental group(siRNA LHPP group),and real-time fluorescence quantitative PCR and Western blotting were used to verify the interference effect of small interfering RNA(siRNA LHPP).The effects of LHPP interference on U87MG cell proliferation and apoptosis were detected using CCK8 method and flow cytometry.Transwell method was used to compare the changes in the migration ability of U87MG cells in the experimental group.Western blotting was used to detect the effect of LHPP interference on proteins related to the transforming growth factorβ/Smad 2(TGF-β/Smad2)pathway in U87MG cells.Results Bioinformatics results showed that LHPP transcription and expression decreased in high-grade gliomas,leading to shortened patient survival(all P<0.001).Compared to the negative control group,the LHPP mRNA transcription(t=9.090,P<0.001)and protein expression levels(t=9.568,P<0.001)in the experimental group cells were down regulated by 60%;the proliferation ability of the experimental group cells increased by about 50%,and the migration ability increased by about 100%,with the significant increase in the expression of Ki-67,TGF-βand p-Smad2.Conclusion The absence of LHPP activated TGF-β/Smad2 signaling pathway,and promoted the proliferation and metastasis of glioblastoma,leading to poor prognosis.