Key Laboratory Experimental, Teratology of the Ministry of Education, Department of Biochemistry and Molecular Biology, Shandong University School of Basic Medical Sciences, 44 Wenhua Xi Road, Jinan, 250012, Shandong, China.
Shandong Medical College, Linyi, China.
Cell Oncol (Dordr). 2020 Jun;43(3):395-407. doi: 10.1007/s13402-020-00494-9. Epub 2020 Jan 31.
FAM83D has been proposed to act as an oncoprotein in several types of human cancer. Its role and mode of action in human non-small cell lung cancer (NSCLC) metastasis and its impact on chemotherapy are as yet, however, poorly understood.
FAM83D expression was measured in NSCLC cells and normal lung epithelial cells, as well as in primary NSCLC tissues and corresponding adjacent non-cancerous tissues, using qRT-PCR, Western blotting and immunohistochemistry. FAM83D was stably overexpressed in BEAS2B cells or silenced in A549 and H1299 cells using retroviral or lentiviral vectors. The growth capacity of NSCLC cells was evaluated using MTT and colony formation assays. Epithelial-mesenchymal transition (EMT) was assessed using Western blotting and immunofluorescence. NSCLC cell invasive capacities were assessed using scratch wound healing and Boyden chamber assays. NSCLC cell viability in response to cisplatin treatment was assessed using MTT assays in vitro and a xenograft model in vivo.
We found that FAM83D expression levels were significantly elevated in NSCLC cells and tissues, and positively correlated with tumor progression and a poor prognosis. Exogenous FAM83D overexpression promoted, while FAM83D silencing inhibited NSCLC cell proliferation, EMT and invasion. FAM83D silencing also reduced cisplatin resistance. Concordantly, we found that NSCLC patients with a low FAM83D expression benefited most from chemotherapy. Mechanistically, we found that FAM83D activated the protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway. Pharmacological treatment with either AKT or mTOR inhibitors reverted FAM83D-induced tumorigenic phenotypes.
Our results suggest a role of FAM83D in NSCLC development. In addition, our results indicate that NSCLC patients exhibiting FAM83D overexpression are likely to benefit from AKT and/or mTOR inhibitor treatment.
FAM83D 已被提出在几种类型的人类癌症中作为癌蛋白发挥作用。然而，其在人类非小细胞肺癌（NSCLC）转移中的作用和作用方式及其对化疗的影响仍知之甚少。
使用 qRT-PCR、Western blot 和免疫组织化学检测 FAM83D 在 NSCLC 细胞和正常肺上皮细胞以及原发性 NSCLC 组织和相应的相邻非癌组织中的表达。使用逆转录病毒或慢病毒载体在 BEAS2B 细胞中稳定过表达 FAM83D 或在 A549 和 H1299 细胞中沉默 FAM83D。使用 MTT 和集落形成测定评估 NSCLC 细胞的生长能力。使用 Western blot 和免疫荧光评估上皮-间充质转化（EMT）。使用划痕愈合和 Boyden 室测定评估 NSCLC 细胞的侵袭能力。使用 MTT 测定法体外和异种移植模型体内评估 NSCLC 细胞对顺铂治疗的敏感性。
我们发现 FAM83D 表达水平在 NSCLC 细胞和组织中显著升高，并且与肿瘤进展和预后不良呈正相关。外源性 FAM83D 过表达促进，而 FAM83D 沉默抑制 NSCLC 细胞增殖、EMT 和侵袭。FAM83D 沉默也降低了顺铂耐药性。一致地，我们发现 FAM83D 低表达的 NSCLC 患者从化疗中获益最大。机制上，我们发现 FAM83D 激活蛋白激酶 B（AKT）/哺乳动物雷帕霉素靶蛋白（mTOR）通路。用 AKT 或 mTOR 抑制剂进行药理学治疗可逆转 FAM83D 诱导的致瘤表型。
我们的研究结果表明 FAM83D 在 NSCLC 发展中起作用。此外，我们的结果表明，FAM83D 过表达的 NSCLC 患者可能受益于 AKT 和/或 mTOR 抑制剂治疗。
