Li Xia, Sun Cui, Chen Jing, Ma Ji-Fen, Pan Yi-Heng
Center for Diagnosis and Treatment of Neuro-oncology Diseases, Taihe Hospital, Hubei University of Medicine, Shiyan 442000, Hubei, China.
Center for Diagnosis and Treatment of Neuro-oncology Diseases, Taihe Hospital, Hubei University of Medicine, Shiyan 442000, Hubei, China.
Transl Oncol. 2022 Aug;22:101454. doi: 10.1016/j.tranon.2022.101454. Epub 2022 May 23.
To explore the mechanism by which the family with sequence similarity 83, member D (FAM83D)-mediated AKT/mTOR signaling pathway activation affects the proliferation and metastasis of glioma cells.
FAM83D protein expression in glioma cells and tissues was detected by western blotting. Glioma U87 and U251 cells were selected and divided into the Mock, siNC, siFAM83D, FAM83D, MK2206 and FAM83D + MK2206 groups. Cell proliferation was assessed by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) and clone formation assays, while invasion and migration were evaluated by Transwell assays and wound healing tests. The protein expression of members of the AKT/mTOR pathway was determined via western blotting. Xenograft models were also established in nude mice to observe the in vivo effect of FAM83D on the growth of glioma.
FAM83D was upregulated in glioma patients, especially in those with Stage III-IV. In addition, cells treated with siFAM83D had significant downregulation of p-AKT/AKT and p-mTOR/mTOR, with decreased proliferation and colony numbers, as well as decreased invasion and migration compared to the Mock group. However, FAM83D overexpression could activate the Akt/mTOR pathway and promote the proliferation, invasion and migration of glioma cells. Moreover, treatment with MK2206, an inhibitor of AKT, reversed the promoting effect of FAM83D on the growth of glioma cells. The in vivo experiments demonstrated that silencing FAM83D could inhibit the in vivo growth of glioma cells CONCLUSION: FAM83D was upregulated in glioma and silencing FAM83D suppressed the proliferation, invasion and migration of glioma cells via inhibition of the AKT/mTOR pathway.
探讨序列相似性家族83成员D（FAM83D）介导的AKT/mTOR信号通路激活影响胶质瘤细胞增殖和转移的机制。
采用蛋白质免疫印迹法检测胶质瘤细胞和组织中FAM83D蛋白表达。选取胶质瘤U87和U251细胞，分为Mock组、siNC组、siFAM83D组、FAM83D组、MK2206组和FAM83D + MK2206组。通过MTT（3-[4,5-二甲基噻唑-2-基]-2,5-二苯基溴化四氮唑）法和克隆形成实验评估细胞增殖，通过Transwell实验和伤口愈合实验评估侵袭和迁移能力。采用蛋白质免疫印迹法检测AKT/mTOR通路成员的蛋白表达。还在裸鼠中建立异种移植模型，观察FAM83D对胶质瘤生长的体内作用。
FAM83D在胶质瘤患者中上调，尤其是在Ⅲ-Ⅳ期患者中。此外，与Mock组相比，用siFAM83D处理的细胞中p-AKT/AKT和p-mTOR/mTOR显著下调，增殖和集落数量减少，侵袭和迁移能力降低。然而，FAM83D过表达可激活Akt/mTOR通路，促进胶质瘤细胞的增殖、侵袭和迁移。此外，用AKT抑制剂MK2206处理可逆转FAM83D对胶质瘤细胞生长的促进作用。体内实验表明，沉默FAM83D可抑制胶质瘤细胞的体内生长。结论：FAM83D在胶质瘤中上调，沉默FAM83D可通过抑制AKT/mTOR通路抑制胶质瘤细胞的增殖、侵袭和迁移。
