目的：探讨miR-330-3p/SLC7A11在干扰素-γ（interferon-γ,IFN-γ）调控口腔鳞状细胞癌铁死亡中的功能和机制。方法：培养人口腔鳞状细胞癌（oral squamous cell carcinoma,OSCC）细胞系,分为对照组和IFN-γ组。CCK8、集落形成测定、细胞侵袭实验和流式细胞术检测细胞活力、迁徙、侵袭和凋亡。qPCR和蛋白质印迹法测量OSCC细胞中miR-330-3p、SLC7A11表达,靶向位点由TargetScan预测,并通过双荧光素酶报告测定法确认,通过操纵miR-330-3p和SLC7A11在OSCC细胞系中的表达水平。铁、ROS水平分析和裸鼠研究检测铁死亡和IFN-γ调控OSCC细胞在体内的生长。结果：与对照组比较,IFN-γ显着降低了OSCC细胞活力,抑制了OSCC细胞的增殖、迁移和侵袭;IFN-γ抑制了OSCC细胞中铁死亡阴性调节因子的水平,包括GPX4和SLC7A11,IFN-γ诱导的ROS、铁和Fe2+在OSCC细胞中积累。通过荧光素酶报告法验证靶向关系,miR-330-3p可有效抑制OSCC细胞中SLC7A11水平。miR-330-3p模拟物增加了OSCC细胞迁移,并降低了细胞凋亡和铁死亡。而裸鼠的致瘤性测定表明,IFN-γ抑制了OSCC细胞在体内的生长。结论：miR-300-3p降低SLC7A11的表达促进IFN-γ诱导的OSCC细胞的增殖和侵袭,抑制细胞凋亡和铁死亡。
Abstract
Objective To investigate the function and mechanism of miR-330-3p/SLC7A11 in the regulation of proliferation, apoptosis and ferroptosis in oral squamous cell carcinoma by interferon-γ. Methods Human OSCC cell lines were cultured and divided into control group and IFN-γgroup. Cell viability, migration, invasion, and apoptosis were measured by CCK8, colony formation assay, cell invasion assay, and flow cytometry. The expression of miR-330-3p and SLC7A11 in OSCC cells was measured by qPCR and Western blotting. The target sites of miR-330-3p were predicted by TargetScan and confirmed by dual luciferase reporting assay, which manipulated the expression levels of miR-330-3p and SLC7A11 in OSCC cell lines. Ferroptosis and IFN-γ regulation of OSCC cell growth in vivo were detected by iron and ROS level analysis and nude mouse study. Results Compared with the control group, IFN-γsignificantly reduced OSCC cell viability and inhibited the proliferation, migration and invasion of OSCC cells. IFN-γinhibited levels of negative regulators of iron apoptosis in OSCC cells, including GPX4 and SLC7A11, IFN-γ-induced ROS, and accumulation of iron and Fe2+ in OSCC cells. The targeting relationship was verified by luciferase reporting method, and miR-330-3p effectively inhibited SLC7A11 levels in OSCC cells. MiR-330-3p mimics increased OSCC cell migration and decreased apoptosis and ferroptosis. Tumogenicity in nude mice showed that IFN-γinhibited OSCC cell growth in vivo. Conclusion MiR-300-3p inhibits the expression of SLC7A11 to promote IFN-γ-induced proliferation and invasion of OSCC cells, and inhibit cell apoptosis and ferroptosis.
关键词
miR-300-3p /
SLC7A11 /
干扰素-γ /
口腔鳞状细胞癌 /
铁死亡
Key words
miR-300-3p /
SLC7A11 /
IFN-γ /
oral squamous cell carcinoma /
ferroptosis
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] PANARESE I, AQUINO G, RONCHI A, et al.Oral and Oropharyngeal Squamous Cell Carcinoma: Prognostic and Predictive Parameters in the Etiopathogenetic Route[J]. Expert Rev Anticancer Ther, 2019, 19: 105-119.
[2] LI J, CAO F, YIN HL, et al.Ferroptosis: Past, Present and Future[J]. Cell Death Dis, 2020, 11: 88.
[3] INUI M, MARTELLO G, PICCOLO S.MicroRNA control of signal transduction[J]. Nat Rev Mol Cell Biol, 2010, 11(4): 252-263.
[4] SUN D, LI X, MA M, et al.The predictive value and potential mechanisms of miRNA-328 and miRNA-378 for brain metastases in operable and advanced non-small-cell lung cancer[J]. Jpn J Clin Oncol, 2015, 45(5): 464-473.
[5] YAN LX, HUANG XF, SHAO Q, et al.MicroRNA miR-21 overexpression in human breast cancer is associated with advanced clinical stage, lymph node metastasis and patient poor prognosis[J]. RNA, 2008, 14(11): 2348-2360.
[6] LIU MK, MA T, YU Y, et al.MiR-1/GOLPH3/Foxo1 Signaling Pathway Regulates Proliferation of Bladder Cancer[J]. Technol Cancer Res Treat, 2019, 18: 1533033819886897.
[7] LIN W, WANG C, LIU G, et al.SLC7A11/xCT in cancer: biological functions and therapeutic implications[J]. Am J Cancer Res, 2020, 10(10): 3106-3126.
[8] ZHANG P, WANG W, WEI Z, et al.xCT expression modulates cisplatin resistance in Tca8113 tongue carcinoma cells[J]. Oncol Lett, 2016, 12(1): 307-314.
[9] GAO AH, HU YR, ZHU WP.IFN-γ inhibits ovarian cancer progression via SOCS1/JAK/STAT signaling pathway[J]. Clin Transl Oncol, 2022, 24(1): 57-65.
[10] YONEKURA N, YOKOSAWA N, HARIYA Y, et al.Interferon-alpha and interferon-gamma augment apoptosis in oral carcinoma cells treated with Shiga toxin 2(Stx2)[J]. Tumor Res, 2002, 35: 25-33.
[11] CHEN, D, YAN W, LIU Z, et al.Downregulation of miR‐221 enhances the sensitivity of human oral squamous cell carcinoma cells to Adriamycin through upregulation of TIMp3 expression[J]. Biomed Pharmacother, 2016, 77: 72-78.
[12] ZENG Z, VEITCH M, KELLY GA, et al.IFN-γ Critically Enables the Intratumoural Infiltration of CXCR3+ CD8+ T Cells to Drive Squamous Cell Carcinoma Regression[J]. Cancers (Basel), 2021, 13(9): 2131.
[13] ZHOU Z H, ZHAO T C, LIANG S Y, et al.A therapeutic approach with combination of interferon-gamma and autophagy inhibitor for oral squamous cell carcinoma[J]. Am J Cancer Res, 2021, 11(4): 1503-1521.
[14] WEI TT, ZHANG MY, ZHENG XH, et al.Interferon-γ induces retinal pigment epithelial cell Ferroptosis by a JAK1-2/STAT1/SLC7A11 signaling pathway in Age-related Macular Degeneration[J]. FEBS J, 2022, 289(7): 1968-1983.
[15] LI Q, WANG W, ZHANG M, et al.Circular RNA circ-0016068 promotes the growth, migration, and invasion of prostate cancer cells by regulating the miR-330-3p/BMI-1 axis as a competing endogenous RNA[J]. Front Cell Dev Biol, 2020, 8: 827.
[16] CHENG Y, ZHU H, GAO W.MicroRNA-330-3p represses the proliferation and invasion of laryngeal squamous cell carcinoma through downregulation of Tra2β-mediated Akt signaling[J]. Mol Cell Probes, 2020, 52: 101574.
[17] QIAN C, CHEN S, LI S, et al.Circ_0000003 regulates glutamine metabolism and tumor progression of tongue squamous cell carcinoma via the miR-330-3p/GLS axis[J]. Oncol Rep, 2021, 45(4): 45.
[18] MUKHOPADHYAY S, BIANCUR DE, PARKER SJ, et al.Autophagy is Required for Proper Cysteine Homeostasis in Pancreatic Cancer Through Regulation of SLC7A11[J]. Proc Natl Acad Sci USA, 2021, 118(6): e2021475118.
基金
2021年保定市科技计划“SLC7A11-AS1调控miR-429对口腔鳞癌细胞CAL-27细胞增殖、凋亡及迁移的影响” （2141ZF019）; 2020年河北省卫健委重点科技研究计划“miR-330-3pSLC7A11表达在口腔鳞状细胞癌铁死亡中的作用”（20201256）
PDF(3432 KB)
