miR-29a对胶质母细胞瘤细胞增殖、迁移、侵袭及PTEN/AKT/GSK-3β通路的影响-现代医学

miR-29a对胶质母细胞瘤细胞增殖、迁移、侵袭及PTEN/AKT/GSK-3β通路的影响

单位：河北北方学院附属第一医院神经外科, 河北张家口 075000

关键词：miR-29a 胶质母细胞瘤细胞增殖迁移侵袭人张力蛋白同源物基因/蛋白激酶B/糖原合成酶通路

分类号：R730.264

出版年·卷·期（页码）：2021·49·第八期（884-892）

摘要：

目的：研究miR-29a对胶质母细胞瘤（GBM）细胞增殖、迁移、侵袭及人张力蛋白同源物基因/蛋白激酶B/糖原合成酶（PTEN/AKT/GSK-3β）通路的影响。方法：体外培养GBM细胞系TG-905，分为空白对照组（NG组）、阴性对照组（mimics-NC组）、miR-29a过表达组（miR-29a-mimics组）和PTEN抑制剂组（miR-29a-mimics+Bpv组）。CCK-8法、平板克隆实验检测各组细胞增殖情况；划痕实验、Transwell实验分别检测各组细胞迁移、侵袭能力；免疫印迹法检测细胞增殖相关核抗原（PCNA）、迁移及侵袭相关蛋白[基质金属蛋白酶9（MMP-9）、E-钙黏附蛋白（E-cadherin）、波形蛋白（Vimentin）、N-钙黏附蛋白（N-cadherin）]及通路相关蛋白（PTEN、p-PI3K、PI3K、p-AKT、AKT、GSK-3β、β-catenin）表达变化。结果：细胞转染成功；与NG组、mimics-NC组比较，miR-29a-mimics组TG-905细胞增殖抑制率及PTEN、E-Cadherin、PTEN、GSK-3β蛋白表达显著升高（P<0.05），细胞克隆形成率、划痕愈合率、侵袭数及Ki-67、MMP-9、N-Cadherin、Vimentin蛋白表达显著降低或减少（P<0.05）；与miR-29a-mimics组比较，miR-29a-mimics+Bpv组TG-905细胞增殖抑制率及PTEN、E-Cadherin、PTEN、GSK-3β蛋白表达显著降低（P<0.05），细胞克隆形成率、划痕愈合率、侵袭数及Ki-67、MMP-9、N-Cadherin、Vimentin蛋白表达显著升高或增加（P<0.05）。结论：过表达miR-29a可能通过上调PTEN表达抑制PI3K/Akt/GSK-3β信号通路活化抑制TG-905细胞增殖、侵袭及迁移。

Objective: To study the effects of miR-29a on cell proliferation, migration, invasion and phosphatase and tensin homologue-deleted chromosome ten gene/protein kinase B/glycogen synthase kinase-3β(PTEN/AKT/GSK-3β) pathway of glioblastoma multiforme cells(GBM).Methods: GBM cell line TG-905 was cultured in vitro and divided into blank control group(NG group), negative control group(mimics-NC group), miR-29a overexpression group(miR-29a-mimics group), and PTEN inhibitor group(miR-29a-mimics+Bpv group). CCK-8 method and plate cloning experiment were used to detect cell proliferation in each group; Scratch test and Transwell test were used to detect cell migration and invasion ability of each group respectively; Western blotting was used to detect cell proliferating cell nuclear antigen(PCNA), migration and invasion-related proteins such as matrix metalloproteinase 9(MMP-9), E-cadherin, Vimentin, N-cadherin,as well as pathway related protein PTEN, p-PI3K, PI3K, p-AKT, AKT, GSK-3β, β-catenin.Results: The cells were successfully transfected; compared with the NG group and the mimics-NC group, the TG-905 cell proliferation inhibition rate and PTEN, E-Cadherin, PTEN, GSK-3β proteins expression were significantly increased in miR-29a-mimics group(P<0.05), the cell clone formation rate, scratch healing rate, invasion number, and Ki-67, MMP-9, N-Cadherin, and Vimentin proteins expression were significantly reduced(P<0.05); compared with miR-29a-mimics group, the TG-905 cell proliferation inhibition rate and PTEN, E-Cadherin, PTEN, GSK-3β proteins expression were significantly reduced in miR-29a-mimics+Bpv group(P<0.05), the cell clone formation rate, scratch healing rate, invasion number, and Ki-67, MMP-9, N-Cadherin, and Vimentin proteins expression were significantly increased(P<0.05). Conclusion: Overexpression of miR-29a may up-regulate the expression of PTEN to inhibit the activation of PI3K/Akt/GSK-3β signaling pathway and inhibit the proliferation, invasion and migration of TG-905 cells.

参考文献：

[1] 孙远召, 杨天明.阿霉素纳米粒对颅内移植G422小鼠的化疗实验研究[J]. 东南大学学报(医学版), 2006(5):341-345.
[2] DAVIDE B, ANGELA C, MATILDE C, et al. CircSMARCA5 inhibits migration of glioblastoma multiforme cells by regulating a molecular axis involving splicing factors SRSF1/SRSF3/PTB[J]. Int J Mol Sci, 2018, 19(2):480-486.
[3] LOU W, LIU J, DING B, et al. Identification of potential miRNA-mRNA regulatory network contributing to pathogenesis of HBV-related HCC[J]. J Transl Med, 2019, 17(1):7-13.
[4] 张利超, 卢忠胜, 张强.miR-29a过表达对胶质瘤细胞迁移的作用及机制[J]. 山东医药, 2018, 58(37):52-56.
[5] SONG Z, HAN X, SHEN L, et al. PTEN silencing enhances neuronal proliferation and differentiation by activating PI3K/Akt/GSK3β pathway in vitro[J]. Exp Cell Res, 2018, 363(2):179-187.
[6] 刘学键, 武霞, 李玉花.通过PI3K/AKT途径调节胶质母细胞瘤CD47表达对肿瘤侵袭性的影响[J]. 实用临床医药杂志, 2020, 24(7):56-61.
[7] 梁媛, 冯洋洋, 李琳琳, 等.MicroRNA-29a靶向抑制PTEN基因诱导非小细胞肺癌细胞上皮间质转化的机制研究[J]. 现代肿瘤医学, 2018, 65(5):653-659.
[8] 孙梦雪, 莫海亚, 何晓璞, 等.miR-29a通过调控PTEN/AKT信号通路对CCl4诱导的肝纤维化大鼠的影响[J]. 国际消化病杂志, 2020, 40(1):26-30.
[9] 贾廷印, 李好朝, 乔泽强, 等.蛇床子素通过上调抑癌基因PTEN抑制肝癌细胞HepG2增殖并促进细胞凋亡[J]. 中国免疫学杂志, 2019, 35(2):192-196.
[10] DENG J, ZHOU Y, LONG W.Analysis of gender-specific regulatory mechanisms on the oncogenesis and prognosis of glioblastoma multiforme[J]. J Phys Conf Ser, 2020, 1575(666):12059-12065.
[11] HU X, TAN S, YIN H, et al. Selenium-mediated gga-miR-29a-3p regulates LMH cell proliferation, invasion, and migration by targeting COL4A2[J]. Metallomics, 2020, 12(3):1-10.
[12] 赵喻.miR-29a诱导脑胶质瘤细胞凋亡的分子机制[D].上海:华东师范大学, 2010.
[13] HAN Y, SUN G.Overexpression of lncRNA TINCR is associated with high-grade, invasive, and recurring tumors, and facilitates proliferation in vitro and in vivo of urothelial carcinoma of the bladder[J]. Urolo Oncol-Semin Ori, 2020, 12(27):1-8.
[14] QIU J J, GUO J J, LV T J, et al Prognostic value of centromere protein-a expression in patients with epithelial ovarian cancer[J]. Tumor Biol, 2013, 34(5):1-8.
[15] 刘梦瑶, 苟理尧, 夏菁, 等.整合素β2促进人乳腺癌细胞MCF-7的迁移, 侵袭与粘附[J]. 基因组学与应用生物学, 2019, 38(1):323-329.
[16] 高兵, 陈翰卿, 时正鹏, 等.MiR-29a下调共刺激分子B7-H3的表达及其对脑胶质瘤细胞侵袭的影响[J]. 中国肿瘤生物治疗杂志, 2015, 22(1):28-33.
[17] 于艳, 赵万超, 许烨, 等.盐酸小檗碱对慢性肾衰大鼠肾间质纤维化改善作用及对PTEN/PI3K/AKT信号通路影响[J]. 辽宁中医药大学学报, 2020, 191(3):32-36.
[18] ZHAO S J, KONG F Q, JIE J, et al. Macrophage MSR1 promotes BMSC osteogenic differentiation and M2-like polarization by activating PI3K/AKT/GSK3β/β-catenin pathway[J]. Theranostics, 2020, 10(1):17-35.
[19] HAO Q, GAO L, NIU W, et al. POTEE stimulates the proliferation of pancreatic cancer by activating the PI3K/Akt/GSK-3β/β-catenin signaling[J]. BioFactors, 2020, 46(4):1-6.
[20] ZHANG Y, CHENG H, LI W, et al. Highly-expressed P2X7 receptor promotes growth and metastasis of human HOS/MNNG osteosarcoma cells via PI3K/Akt/GSK3β/β-catenin and mTOR/HIF1α/VEGF signaling[J]. Int J Cancer, 2019, 145(4):1068-1082.
[21] SHI X, VALIZADEH A, MIR S M, et al. miRNA-29a reverses P-glycoprotein-mediated drug resistance and inhibits proliferation via up-regulation of PTEN in colon cancer cells[J]. Eur J Pharmacol, 2020, 880(122):136-143.

服务与反馈：

【文章下载】【发表评论】【查看评论】【加入收藏】

提示：您还未登录，请登录！点此登录