TMEM106B通过Her2信号通路影响脑膜瘤细胞增殖和血管生成的研究

发布时间：2018-06-29 13:47

本文选题：恶性脑膜瘤 + TMEM106B　；参考：《南昌大学》2017年硕士论文

【摘要】：目的:观察TMEM106B基因是否影响恶性脑膜瘤细胞的增殖和血管生成能力,并讨论TMEM106B是否通过Her2信号通路发挥作用。方法:1、通过病毒感染沉默TMEM106B基因质粒至恶性脑膜瘤IOMM-Lee和CH157细胞株中。实验分组:第一组:IOMM-Lee空白组(IOMM-Lee-blank);第二组:IOMM-Lee沉默阴性对照组(IOMM-Lee-NC-sh);第三组:IOMM-Lee沉默TMEM106B基因组(IOMM-Lee-TMEM106B-sh);第四组:CH157空白组(CH157-blank);第五组:CH157沉默阴性对照组(CH157-NC-sh);第六组CH157沉默TMEM106B基因组(CH157-TMEM106B-sh)。使用q-PCR、Western blot检测各组细胞TMEM106B mRNA和蛋白表达水平。2、MTT、EdU法检测各组细胞增殖情况;流式细胞术检测各组细胞周期变化情况;小管形成实验检测各组细胞血管生成情况。3、q-PCR和Western blot检测各组细胞Her2、JUN和VEGF mRNA和蛋白表达水平。4、应用Her2抑制剂Afatinib(BIBW2992)干扰恶性脑膜瘤细胞IOMM-Lee和CH157后,q-PCR和Western blot检测Her2、JUN和VEGF mRNA和蛋白表达水平。结果:1、慢病毒转染成功,TMEM106B在mRNA和蛋白水平达到分组要求。2、脑膜瘤细胞株IOMM-Lee和CH157沉默TMEM106B组细胞增殖和血管生成能力均减弱;TMEM106B基因通过影响细胞周期中的G0/G1-S的变化而起作用。3、沉默TMEM106B基因后,恶性脑膜瘤细胞IOMM-Lee和CH157的Her2和JUN基因m RNA和蛋白表达无明显变化。4、干扰Her2基因后,恶性脑膜瘤细胞IOMM-Lee和CH157的TMEM106B和JUN基因m RNA和蛋白表达均减少。结论:1、TMEM106B可以促进恶性脑膜瘤细胞株IOMM-Lee和CH157的增殖和血管生成;2、TMEM106B可能通过Her2信号通路影响恶性脑膜瘤细胞增殖和血管生成。
[Abstract]:Aim: to investigate whether TMEM106B gene affects the proliferation and angiogenesis of malignant meningioma cells and discuss whether TMEM106B plays a role through Her2 signaling pathway. Methods the TMEM106B gene plasmid was silenced by virus infection into IOMM-Lee and CH157 cell lines of malignant meningioma. The first group: IOMM-Lee-blank; the second group: IOMM-Lee-NC-sh; the third group: IOMM-Lee silencing TMEM106B (IOMM-Lee-TMEM106B-sh); the fourth group: CH157-blank; the fifth group: CH157-NC-sh; the sixth group CH157 silencing TMEM106B (CH157-TMEM106B-sh). The expression level of TMEM106B mRNA and protein was detected by q-PCRX Western blot. The proliferation of TMEM106B cells in each group was detected by TMEM106B mRNA and protein expression, and the cell cycle was detected by flow cytometry. The expression of Her2JUN and VEGF mRNA and protein in each group was detected by tubulogenesis assay and Western blot. Her2 inhibitor Afatinib (BIBW2992) was used to interfere with malignant meningioma cell line IOMM-Lee and CH157, and Western blot was used to detect Her2JUN and VEGFmRNA. And protein expression level. Results TMEM106B was successfully transfected with lentivirus to meet the requirement of grouping at mRNA and protein levels. The proliferation and angiogenesis ability of TMEM106B cells in meningioma cell lines IOMM-Lee and CH157 silenced by affecting the changes of G0 / G1-S in cell cycle was decreased. After silencing TMEM106B gene, The mRNA and protein expression of Her2 and Jun genes in malignant meningioma cell lines IOMM-Lee and CH157 showed no significant change. After interfering with Her2 gene, the mRNA and protein expressions of TMEM106B and Jun genes in IOMM-Lee and CH157 were decreased. Conclusion TMEM106B can promote the proliferation and angiogenesis of malignant meningioma cell lines IOMM-Lee and CH157. TMEM106B may affect the proliferation and angiogenesis of malignant meningioma cells through Her2 signaling pathway.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.45

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