Pituitary adenomas（PAs） are well known as a common intracranial benign tumor, and a portion of PAs are refractory to current therapeutic methods. Erb B receptors family signaling pathway regulates the expression of ...Pituitary adenomas（PAs） are well known as a common intracranial benign tumor, and a portion of PAs are refractory to current therapeutic methods. Erb B receptors family signaling pathway regulates the expression of PAs activation associated gene. Inhibition of epidermal growth factor receptor（EGFR） can inhibit proliferation of PAs. Leucine-rich repeats and immunoglobulin-like domains protein 1（ LRIG1）, a negative mediated gene of Erb B receptors family, plays a role in many tumors. However, there are seldom researches about the functional role of LRIG1 in PAs. The aim of this study is to explore the potential effect of LRIG1 and its regulating mechanism in PAs. First, we investigated the role of LRIG1 in cell migration, invasion of PAs with transfected LRIG1 or control. Then, we explored its impact on cell proliferation and apoptosis of PAs in vivo. To study the regulating mechanism of LRIG1, we examined the expression of molecular factor of PI3K/AKT and Ras/Raf/ERK pathway using Western blotting in vitro and RT-PCR in vitro and in vivo. It was found that LRIG1 over-expression inhibited cell migration, invasion and proliferation, and promoted apoptosis of PAs in vivo and in vitro. Furthermore, LRIG1 suppressed the expression of signaling of PI3K/AKT and Ras/Raf/ERK pathways in PAs. LRIG1, as a negative mediated gene of tumor, can inhibit biological function of PAs via inhibiting PI3K/AKT and Ras/Raf/ERK pathways, and it might be a new target for gene therapy of PAs.展开更多
目的探讨过表达多亮氨酸重复区免疫球蛋白样蛋白1(LRIG1)通过细胞毒性T淋巴细胞相关蛋白4(CTLA-4)调控磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(Akt)信号通路影响神经胶质瘤细胞增殖、迁徙和侵袭及其分子机制。方法选取南昌大学第二附属医院201...目的探讨过表达多亮氨酸重复区免疫球蛋白样蛋白1(LRIG1)通过细胞毒性T淋巴细胞相关蛋白4(CTLA-4)调控磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(Akt)信号通路影响神经胶质瘤细胞增殖、迁徙和侵袭及其分子机制。方法选取南昌大学第二附属医院2018年9月至2020年6月手术治疗的103例患者的神经胶质瘤组织和癌旁组织为研究对象,癌旁组织为对照组。培养胶质瘤U251细胞,采用荧光定量聚合酶链反应(PT-qPCR)、蛋白质印迹法(Western blot)检测LRIG1、CTLA-4基因及蛋白的表达水平。利用Flag-LRIG1质粒构建LRIG1过表达细胞模型,命名为对照组和LRIG1组。采用细胞计数试剂盒(CCK-8)检测两组细胞增殖能力,细胞划痕及Transwell实验检测两组细胞迁徙和侵袭能力。Western blot检测PI3K、Akt蛋白表达水平。组间数据比较采用t检验。结果胶质瘤组LRIG1表达水平(1.54±0.11)明显低于癌旁对照组LRIG1表达水平(2.11±0.12),差异有统计学意义(t=6.070,P<0.01)。胶质瘤组CTLA-4表达水平(3.82±0.09)明显高于癌旁对照组CTLA-4表达水平(0.51±0.48),差异有统计学意义(t=8.190,P<0.01)。Flag-LRIG1质粒转染后LRIG1组LRIG1蛋白表达水平(2.83±0.17)高于对照组LRIG1表达水平(1.41±0.08),差异有统计学意义(t=13.090,P<0.01)。LRIG1组细胞吸光度值(0.51±0.04)低于对照组细胞吸光度值(1.11±0.09),差异有统计学意义(t=10.550,P<0.01);划痕实验24 h LRIG1组细胞迁移面积[(9.52±0.12)%]低于对照组细胞迁移面积[(12.05±0.54)%],差异有统计学意义(t=7.920,P<0.05);Transwell实验LRIG1组细胞迁移数量[(43.15±5.17)个]低于对照组细胞迁移数量[(70.13±6.13)个],差异有统计学意义(t=5.830,P<0.01)。LRIG1组CTLA-4蛋白表达水平(0.50±0.08)低于对照组CTLA-4蛋白表达水平(0.83±0.14),差异有统计学意义(t=3.550,P<0.05)。结论LRIG1通过下调CTLA-4表达来调控PI3K/AKT通路从而抑制脑质瘤细胞的增殖和迁移。展开更多
基金supported by grants from the National Natural Science Foundation of China(No.81560412)Jiangxi Provincial Health Development Planning Commission Project(No.20141065)Jiangxi Provincial Natural Science Foundation of China(No.20152BCB24009 and No.20151BDH80009)
文摘Pituitary adenomas（PAs） are well known as a common intracranial benign tumor, and a portion of PAs are refractory to current therapeutic methods. Erb B receptors family signaling pathway regulates the expression of PAs activation associated gene. Inhibition of epidermal growth factor receptor（EGFR） can inhibit proliferation of PAs. Leucine-rich repeats and immunoglobulin-like domains protein 1（ LRIG1）, a negative mediated gene of Erb B receptors family, plays a role in many tumors. However, there are seldom researches about the functional role of LRIG1 in PAs. The aim of this study is to explore the potential effect of LRIG1 and its regulating mechanism in PAs. First, we investigated the role of LRIG1 in cell migration, invasion of PAs with transfected LRIG1 or control. Then, we explored its impact on cell proliferation and apoptosis of PAs in vivo. To study the regulating mechanism of LRIG1, we examined the expression of molecular factor of PI3K/AKT and Ras/Raf/ERK pathway using Western blotting in vitro and RT-PCR in vitro and in vivo. It was found that LRIG1 over-expression inhibited cell migration, invasion and proliferation, and promoted apoptosis of PAs in vivo and in vitro. Furthermore, LRIG1 suppressed the expression of signaling of PI3K/AKT and Ras/Raf/ERK pathways in PAs. LRIG1, as a negative mediated gene of tumor, can inhibit biological function of PAs via inhibiting PI3K/AKT and Ras/Raf/ERK pathways, and it might be a new target for gene therapy of PAs.
文摘目的探讨过表达多亮氨酸重复区免疫球蛋白样蛋白1(LRIG1)通过细胞毒性T淋巴细胞相关蛋白4(CTLA-4)调控磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(Akt)信号通路影响神经胶质瘤细胞增殖、迁徙和侵袭及其分子机制。方法选取南昌大学第二附属医院2018年9月至2020年6月手术治疗的103例患者的神经胶质瘤组织和癌旁组织为研究对象,癌旁组织为对照组。培养胶质瘤U251细胞,采用荧光定量聚合酶链反应(PT-qPCR)、蛋白质印迹法(Western blot)检测LRIG1、CTLA-4基因及蛋白的表达水平。利用Flag-LRIG1质粒构建LRIG1过表达细胞模型,命名为对照组和LRIG1组。采用细胞计数试剂盒(CCK-8)检测两组细胞增殖能力,细胞划痕及Transwell实验检测两组细胞迁徙和侵袭能力。Western blot检测PI3K、Akt蛋白表达水平。组间数据比较采用t检验。结果胶质瘤组LRIG1表达水平(1.54±0.11)明显低于癌旁对照组LRIG1表达水平(2.11±0.12),差异有统计学意义(t=6.070,P<0.01)。胶质瘤组CTLA-4表达水平(3.82±0.09)明显高于癌旁对照组CTLA-4表达水平(0.51±0.48),差异有统计学意义(t=8.190,P<0.01)。Flag-LRIG1质粒转染后LRIG1组LRIG1蛋白表达水平(2.83±0.17)高于对照组LRIG1表达水平(1.41±0.08),差异有统计学意义(t=13.090,P<0.01)。LRIG1组细胞吸光度值(0.51±0.04)低于对照组细胞吸光度值(1.11±0.09),差异有统计学意义(t=10.550,P<0.01);划痕实验24 h LRIG1组细胞迁移面积[(9.52±0.12)%]低于对照组细胞迁移面积[(12.05±0.54)%],差异有统计学意义(t=7.920,P<0.05);Transwell实验LRIG1组细胞迁移数量[(43.15±5.17)个]低于对照组细胞迁移数量[(70.13±6.13)个],差异有统计学意义(t=5.830,P<0.01)。LRIG1组CTLA-4蛋白表达水平(0.50±0.08)低于对照组CTLA-4蛋白表达水平(0.83±0.14),差异有统计学意义(t=3.550,P<0.05)。结论LRIG1通过下调CTLA-4表达来调控PI3K/AKT通路从而抑制脑质瘤细胞的增殖和迁移。
