National "111" Center for Cellular Regulation and Molecular Pharmaceutics, Key Laboratory of Fermentation Engineering (Ministry of Education), Hubei University of Technology, Wuhan, China.
Membrane Protein Disease Research Group, Department of Physiology, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, AB, Canada.
Autophagy. 2021 Oct;17(10):3175-3195. doi: 10.1080/15548627.2020.1826689. Epub 2020 Oct 22.
Pancreatic cancer is one of the most aggressive tumors associated with a poor clinical prognosis, weakly effective therapeutic options. Therefore, there is a strong impetus to discover new therapeutic targets in pancreatic cancer. In the present study, we first demonstrated that TSPAN1 is upregulated in pancreatic cancer and that TSPAN1 depletion decreases pancreatic cancer cell proliferation and . TSPAN1 expression was correlated with poor overall survival of pancreatic cancer patients. Moreover, we demonstrated that TSPAN1 is a novel positive regulator of macroautophagy/autophagy characterized by decreased LC3-II and SQSTM1/p62 expressions, inhibited puncta formation of GFP-LC3 and autophagic vacuoles. We also demonstrated that mutation impaired autophagy in the zebrafish model. Furthermore, we showed that TSPAN1 promoted autophagy maturation via direct binding to LC3 by two conserved LIR motifs. Mutations in the LIR motifs of TSPAN1 resulted in a loss of the ability to induce autophagy and promote pancreatic cancer proliferation. Second, we discovered two conservative TCF/LEF binding elements present in the promoter region of the gene, which was further verified through luciferase activity and ChIP assays. Furthermore, TSPAN1 was upregulated by FAM83A through the canonical WNT-CTNNB1 signaling pathway. We further demonstrated that both TSPAN1 and FAM83A are both direct targets of (microRNA 454). Additionally, we revealed the role of -FAM83A-TSPAN1 in the proliferation of pancreatic cancer cells and . Our findings suggest that components of the -FAM83A-TSPAN1 axis may be valuable prognosis markers or therapeutic targets for pancreatic cancer.: AMPK: adenosine 5'-monophosphate (AMP)-activated protein kinase; APC: APC regulator of WNT signaling pathway; ATG: autophagy related; AXIN2: axin 2; BECN1: beclin 1; CCND1: cyclin D1; CSNK1A1/CK1α: casein kinase 1 alpha 1; CTNNB1/β-catenin: catenin beta 1; DAPI: 4'6-diamino-2-phenylindole; EBSS: Earle's balanced salt solution; EdU: 5-ethynyl-20-deoxyuridine; FAM83A: family with sequence similarity 83 member A; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GFP: green fluorescent protein; GSEA: gene set enrichment analysis; GSK3B: glycogen synthase kinase 3 beta; IHC: immunohistochemical; LAMP1: lysosomal associated membrane protein 1; LIR: LC3-interacting region; MAP1LC3/LC3, microtubule associated protein 1 light chain 3; : microRNA 454; miRNA: microRNA; MKI67: antigen identified by monoclonal antibody Ki 67; MTOR: mechanistic target of rapamycin kinase; MTT: 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide; MYC: MYC proto-oncogene, bHLH transcription factor; OS: overall survival; PDAC: pancreatic ductal adenocarcinoma; RAB7A: RAB7A, member RAS oncogene family; shRNA: short hairpin RNA; SQSTM1: sequestosome 1; TBE: TCF/LEF binding element; TCGA: The Cancer Genome Atlas; TCF/LEF: transcription factor/lymphoid enhancer binding factor; TCF4: transcription factor 4; TSPAN1: tetraspanin 1; TUNEL: terminal deoxynucleotidyl transferase mediated dUTP nick end labeling; UTR: untranslated region; WT: wild type.
胰腺癌是一种与不良临床预后、疗效差的治疗选择相关的侵袭性肿瘤。因此，在胰腺癌中发现新的治疗靶点的动力很强。在本研究中，我们首先证明 TSPAN1 在胰腺癌中上调，并且 TSPAN1 耗竭降低了胰腺癌细胞的增殖和。TSPAN1 的表达与胰腺癌患者的总生存率差相关。此外，我们证明 TSPAN1 是一种新的自噬/自噬的正调节剂，其特征在于 LC3-II 和 SQSTM1/p62 表达降低，GFP-LC3 和自噬小体的斑点形成受抑制。我们还证明了 突变会损害斑马鱼模型中的自噬。此外，我们表明 TSPAN1 通过与 LC3 的两个保守 LIR 基序直接结合来促进自噬体成熟。TSPAN1 的 LIR 基序突变导致丧失诱导自噬和促进胰腺癌细胞增殖的能力。第二，我们在 基因的启动子区域发现了两个保守的 TCF/LEF 结合元件，通过荧光素酶活性和 ChIP 测定进一步验证。此外，FAM83A 通过经典的 WNT-CTNNB1 信号通路上调 TSPAN1。我们进一步证明，TSPAN1 和 FAM83A 都是 的直接靶标（microRNA 454）。此外，我们揭示了 -FAM83A-TSPAN1 在胰腺癌细胞增殖中的作用 。我们的研究结果表明，-FAM83A-TSPAN1 轴的组成部分可能是胰腺癌有价值的预后标志物或治疗靶点：AMPK：5'-单磷酸腺苷（AMP）激活的蛋白激酶；APC：WNT 信号通路 APC 调节剂；ATG：自噬相关；AXIN2：轴蛋白 2；BECN1：贝塞克林 1；CCND1：细胞周期蛋白 D1；CSNK1A1/CK1α：酪蛋白激酶 1α1；CTNNB1/β-catenin：连接蛋白β 1；DAPI：4'6-二脒基-2-苯基吲哚；EBSS：Earle 的平衡盐溶液；EdU：5-乙炔-20-脱氧尿苷；FAM83A：家族与序列相似性 83 成员 A；GAPDH：甘油醛-3-磷酸脱氢酶；GFP：绿色荧光蛋白；GSEA：基因集富集分析；GSK3B：糖原合成激酶 3β；IHC：免疫组织化学；LAMP1：溶酶体相关膜蛋白 1；LIR：LC3 相互作用区；MAP1LC3/LC3，微管相关蛋白 1 轻链 3；：microRNA 454；miRNA：microRNA；MKI67：由单克隆抗体 Ki 67 鉴定的抗原；MTOR：雷帕霉素靶蛋白激酶；MTT：3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴盐；MYC：MYC 原癌基因，bHLH 转录因子；OS：总生存率；PDAC：胰腺导管腺癌；RAB7A：RAB7A，RAS 癌基因家族成员；shRNA：短发夹 RNA；SQSTM1：自噬体 1；TBE：TCF/LEF 结合元件；TCGA：癌症基因组图谱；TCF/LEF：转录因子/淋巴增强结合因子；TCF4：转录因子 4；TSPAN1：四跨膜蛋白 1；TUNEL：末端脱氧核苷酸转移酶介导的 dUTP 缺口末端标记；UTR：非翻译区；WT：野生型。
