ALG3 contributes to stemness and radioresistance through regulating glycosylation of TGF-β receptor II in breast cancer.
ALG3 contributes to stemness and radioresistance through regulating glycosylation of TGF-β receptor II in breast cancer.
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ALG3 通过调节乳腺癌中 TGF-β 受体 II 的糖基化有助于干性和放射抗性
DOI：
10.1186/s13046-021-01932-8
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发表时间：
2021-04-30
期刊：
影响因子：
--
通讯作者：
Lin H
中科院分区：
文献类型：
--
作者：
Sun X;He Z;Guo L;Wang C;Lin C;Ye L;Wang X;Li Y;Yang M;Liu S;Hua X;Wen W;Lin C;Long Z;Zhang W;Li H;Jian Y;Zhu Z;Wu X;Lin H
放射治疗是乳腺癌的常规和有效的局部治疗方法。然而，由于放射抗性，经常出现残留或复发肿瘤。新的乳腺癌放射抵抗的预测标志物和潜在的治疗靶点需要研究。在本研究中，我们通过RT-PCR筛选了乳腺癌患者样本中的所有10个天冬酰胺连接糖基化（ALG）成员。CCK-8法和流式细胞术检测照射后细胞活力。用殖民地形成试验通过将多靶点单次击中模型拟合到存活分数来确定具有不同ALG 3表达的细胞系的辐射敏感性。通过RT-PCR、Western印迹和流式细胞术评估癌干细胞样特征。Western blot和免疫沉淀法检测ALG 3影响放射敏感性的机制。并在原位异种移植肿瘤模型中验证了ALG 3对IR后肿瘤生长的影响。通过免疫组化证实ALG 3与乳腺癌患者预后的相关性。ALG 3是ALG家族中在放射抵抗乳腺癌组织中过表达最显著的基因。ALG 3过表达预测乳腺癌患者的临床病理特征和总生存期（OS）以及早期局部无复发生存期（LRFS）。在体外和体内，上调ALG 3增强了放射抗性和癌症的干性。相反，ALG 3沉默增加了体外放射敏感性并抑制了癌症的干细胞性，更重要的是，ALG 3的抑制有效地增加了体内乳腺癌细胞的放射敏感性。从机制上讲，我们的结果进一步揭示了ALG 3通过诱导TGF-β受体II（TGFBR 2）的糖基化来促进辐射抗性和癌症的干性。重要的是，使用衣霉素的糖基化衰减和使用LY 2109761的TGFBR 2的抑制都有差异地消除了ALG 3过表达对癌症干性和放射抗性的刺激作用。最后，我们的研究结果表明，放疗在预防ALG 3水平低的乳腺癌患者早期复发方面发挥了重要作用，但在ALG 3过表达的乳腺癌患者中疗效有限。我们的研究结果表明，ALG 3可能作为一个潜在的放射敏感性标志物，并通过调节TGFBR 2的糖基化在乳腺癌中的一个有效的目标，以减少放射抵抗。对于ALG 3水平低的患者，放射仍然是预防乳腺癌早期复发的有效主流疗法。在线版本包含补充材料，可通过10.1186/s13046-021-01932-8获得。
Radiotherapy is a conventional and effective local treatment for breast cancer. However, residual or recurrent tumors appears frequently because of radioresistance. Novel predictive marker and the potential therapeutic targets of breast cancer radioresistance needs to be investigated. In this study, we screened all 10 asparagine-linked glycosylation (ALG) members in breast cancer patients’ samples by RT-PCR. Cell viability after irradiation (IR) was determined by CCK-8 assay and flow cytometry. The radiosensitivity of cell lines with different ALG3 expression was determined with the colony formation assay by fitting the multi-target single hit model to the surviving fractions. Cancer stem-like traits were assessed by RT-PCR, Western blot, and flow cytometry. The mechanisms of ALG3 influencing radiosensitivity was detected by Western blot and immunoprecipitation. And the effect of ALG3 on tumor growth after IR was verified in an orthotopic xenograft tumor models. The association of ALG3 with prognosis of breast cancer patients was confirmed by immunohistochemistry. ALG3 was the most significantly overexpressing gene among ALG family in radioresistant breast cancer tissue. Overexpression of ALG3 predicted poor clinicopathological characteristics and overall survival (OS), and early local recurrence-free survival (LRFS) in breast cancer patients. Upregulating ALG3 enhanced radioresistance and cancer stemness in vitro and in vivo. Conversely, silencing ALG3 increased the radiosensitivity and repressed cancer stemness in vitro, and more importantly inhibition of ALG3 effectively increased the radiosensitivity of breast cancer cells in vivo. Mechanistically, our results further revealed ALG3 promoted radioresistance and cancer stemness by inducing glycosylation of TGF-β receptor II (TGFBR2). Importantly, both attenuation of glycosylation using tunicamycin and inhibition of TGFBR2 using LY2109761 differentially abrogated the stimulatory effect of ALG3 overexpression on cancer stemness and radioresistance. Finally, our findings showed that radiation played an important role in preventing early recurrence in breast cancer patients with low ALG3 levels, but it had limited efficacy in ALG3-overexpressing breast cancer patients. Our results suggest that ALG3 may serve as a potential radiosensitive marker, and an effective target to decrease radioresistance by regulating glycosylation of TGFBR2 in breast cancer. For patients with low ALG3 levels, radiation remains an effective mainstay therapy to prevent early recurrence in breast cancer. The online version contains supplementary material available at 10.1186/s13046-021-01932-8.
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DOI：
10.1158/1078-0432.ccr-12-3408
发表时间：
2013-06-15
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Biddlestone-Thorpe L;Sajjad M;Rosenberg E;Beckta JM;Valerie NC;Tokarz M;Adams BR;Wagner AF;Khalil A;Gilfor D;Golding SE;Deb S;Temesi DG;Lau A;O'Connor MJ;Choe KS;Parada LF;Lim SK;Mukhopadhyay ND;Valerie K
通讯作者：
Valerie K
影响因子：
5.6
作者：
Ferreira IG;Pucci M;Venturi G;Malagolini N;Chiricolo M;Dall'Olio F
通讯作者：
Dall'Olio F
影响因子：
16.6
作者：
Hagey, Daniel W.;Topcic, Danijal;Muhr, Jonas
通讯作者：
Muhr, Jonas
影响因子：
29
作者：
Hwang, In-Young;Kwak, Sojung;Youn, Hong-Duk
通讯作者：
Youn, Hong-Duk
影响因子：
11.2
作者：
Candas D;Lu CL;Fan M;Chuang FY;Sweeney C;Borowsky AD;Li JJ
通讯作者：
Li JJ