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HMGA2对软脑膜转移黑色素瘤细胞迁移和增殖的影响
Effects of HMGA2 on migration and proliferation of leptomeningeal metastatic melanoma
- 中国癌症杂志 2024年34卷第4期 页码：389-399
- 作者机构：
1. 复旦大学脑科学转化研究院，脑功能与脑疾病全国重点实验室，脑科学前沿科学中心，上海 200032
2. 复旦大学附属华山医院神经外科，上海 200040
- 作者简介：
[ "李晓辉（ORCID: 0009-0006-2715-9427），硕士研究生在读。" ]
迟喻丹（ORCID: 0000-0002-4566-3305），研究员。
- 基金信息：国家自然科学基金(32200640)
DOI：10.19401/j.cnki.1007-3639.2024.04.006
中图分类号： R739.45；R739.5收稿：2023-09-21，
修回：2023-12-27，
纸质出版：2024-04-30
- 稿件说明：
移动端阅览
李晓辉, 赵加旭, 彭海豹, 等. HMGA2对软脑膜转移黑色素瘤细胞迁移和增殖的影响[J]. 中国癌症杂志, 2024,34(4):389-399.
Xiaohui LI, Jiaxu ZHAO, Haibao PENG, et al. Effects of HMGA2 on migration and proliferation of leptomeningeal metastatic melanoma[J]. China Oncology, 2024, 34(4): 389-399.
李晓辉, 赵加旭, 彭海豹, 等. HMGA2对软脑膜转移黑色素瘤细胞迁移和增殖的影响[J]. 中国癌症杂志, 2024,34(4):389-399. DOI： 10.19401/j.cnki.1007-3639.2024.04.006.
Xiaohui LI, Jiaxu ZHAO, Haibao PENG, et al. Effects of HMGA2 on migration and proliferation of leptomeningeal metastatic melanoma[J]. China Oncology, 2024, 34(4): 389-399. DOI： 10.19401/j.cnki.1007-3639.2024.04.006.
摘要
背景与目的：
软脑膜转移是黑色素瘤中枢神经系统转移的一种形式，高迁移率族蛋白A2（high-mobility group protein A2，HMGA2）已被证实在多种肿瘤的发生、发展过程中起重要作用，但在软脑膜转移黑色素瘤细胞中的生物学作用尚未明确。本研究在构建黑色素瘤中枢神经系统转移小鼠模型的基础上，探讨软脑膜转移黑色素瘤与原发部位以及脑实质转移黑色素瘤在细胞迁移、细胞增殖等方面的差异，并进一步明确差异表达基因
HMGA
2对软脑膜转移黑色素瘤细胞迁移和增殖的影响。
方法：
通过慢病毒感染构建稳定表达tdTomato和荧光素酶的B16小鼠黑色素瘤细胞（B16-parental cell，B16-Par）。在此基础上，经过体内转移部位适应性筛选，获得B16特异性脑实质转移细胞（B16-brain metastatic cell，B16-BrM）以及B16特异性软脑膜转移细胞（B16-leptomeningeal metastatic cell，B16-LM）。采用细胞划痕实验以及细胞计数试剂盒-8（cell counting kit-8，CCK-8）检测B16-Par、B16-BrM和B16-LM在迁移和增殖能力上的差异。利用转录组测序技术（RNA sequencing，RNA-seq）分析B16-Par、B16-BrM和B16-LM的差异基因表达，筛选出在B16-LM中特异性上调的
HMGA
2基因，用实时荧光定量聚合酶链反应（real-time fluorescence quantitative polymerase chain reaction，RTFQ-PCR）和蛋白质印迹法（Western blot）对该结果进行验证。对B16-LM特异性上调基因进行基因本体论（Gene Ontology，GO）分析。使用siRNA干扰
HMGA
2基因在B16-LM中的表达，通过RTFQ-PCR和Western blot实验对敲低效果进行验证。细胞划痕实验和CCK-8实验检测敲低
HMGA
2对细胞迁移和增殖的影响。利用基因表达综合数据库（Gene Expression Omnibus，GEO）中GSE174401的数据，验证
HMGA
2基因在患者软脑膜转移黑色素瘤细胞中表达的特异性。
结果：
与B16-Par相比，经过脑内环境筛选的肿瘤细胞更易于在中枢神经系统定植。B16-LM具有更强的迁移和增殖能力，且上调表达
HMGA
2基因。GO分析显示，
HMGA
2与血管生成、细胞增殖等多个生物学过程相关。敲低
HMGA
2基因的表达可以抑制B16-LM的迁移和增殖。
HMGA
2在患者软脑膜转移黑色素瘤细胞中表达上调。
结论：
软脑膜转移黑色素瘤细胞具有相对独特的细胞生物学行为，其通过上调
HMGA
2基因的表达来促进细胞迁移和增殖。
Abstract
Background and purpose:
Leptomeningeal metastasis is a form of central nervous system metastasis of melanoma. High mobility group A2 (HMGA2) has been proven to play an important role in th
e occurrence and development of various tumors
but its biological functions in leptomeningeal metastatic melanoma cells remain unclear. On the basis of building mouse models of central nervous system metastasis of melanoma
this study investigated the differences in cell migration and cell proliferation among leptomeningeal metastatic melanoma cells
primary site melanoma cells and brain parenchymal metastatic melanoma cells
and further clarified the effects of differentially expressed gene
HMGA
2 on cell migration and proliferation of leptomeningeal metastatic melanoma cells.
Methods:
B16 mouse melanoma cells (B16-parental cells
B16-Par) stably expressing tdTomato and luciferase were generated by lentiviral infection. Subsequently
B16 specific brain parenchymal metastatic cells (B16-brain metastatic cells
B16-BrM) and B16 specific leptomeningeal metastatic cells (B16-leptomeningeal metastatic cells
B16-LM) were collected after adaptive screening of metastatic sites
in vivo
. The differences in migration and proliferation among B16-Par
B16-BrM and B16-LM were assessed by wound healing assay and cell counting kit-8 (CCK-8). RNA sequencing (RNA-seq) was used to analyze differential gene expression in B16-Par
B16-BrM and B16-LM
and
HMGA
2 gene specifically upregulated in B16-LM was screened out. The results were verified by real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) and Western blot. Gene ontology (GO) analysis was performed for genes which were upregulated in B16-LM specifically. siRNA was used to interfere with the expression of
HMGA
2 gene in B16-LM
and the knock-down effect was verified by RTFQ-PCR and Western blot. The effects of knocking down
HMGA
2 on cell migration and proliferation were detected by wound healing assay and CCK-8 assay. Using GSE174401 data in Gene Expression Omnibus (GEO)
the specificity of
HMGA
2 gene expression in leptomeningeal metastatic me
lanoma cells from patients was verified.
Results:
Compared with Par cells
tumor cells screened by the brain environment were more likely to colonize the central nervous system. B16-LM had stronger migration and proliferation abilities
and upregulated the expression of
HMGA
2 gene. GO analysis revealed that
HMGA
2 was associated with many biological processes such as angiogenesis and cell proliferation. When the expression of
HMGA
2 gene was knocked down
the migration and proliferation of B16-LM could be inhibited.
HMGA
2 was upregulated in leptomeningeal metastatic melanoma cells from patients.
Conclusion:
Leptomeningeal metastatic melanoma cells had relatively unique cellular characteristics
which promoted cell migration and proliferation by upregulating
HMGA
2 gene expression.
关键词
Keywords
references
SCHADENDORF D , VAN AKKOOI A C J , BERKING C , et al . Melanoma [J ] . Lancet , 2018 , 392 ( 10151 ): 971 - 984 . DOI: S0140-6736(18)31559-9 http://doi.org/S0140-6736(18)31559-9
KENCHAPPA R S , TRAN N , RAO N G , et al . Novel treatments for melanoma brain metastases [J ] . Cancer Control , 2013 , 20 ( 4 ): 298 - 306 .
SMALLEY I , CHEN Z H , PHADKE M , et al . Single-cell characterization of the immune microenvironment of melanoma brain and leptomeningeal metastases [J ] . Clin Cancer Res , 2021 , 27 ( 14 ): 4109 - 4125 .
FIFE K M , COLMAN M H , STEVENS G N , et al . Determinants of outcome in melanoma patients with cerebral metastases [J ] . J Clin Oncol , 2004 , 22 ( 7 ): 1293 - 1300 . DOI: 10.1200/JCO.2004.08.140 http://doi.org/10.1200/JCO.2004.08.140
SAMPSON J H , CARTER J H , FRIEDMAN A H , et al . Demographics, prognosis, and therapy in 702 patients with brain metastases from malignant melanoma [J ] . J Neurosurg , 1998 , 88 ( 1 ): 11 - 20 . DOI: 10.3171/jns.1998.88.1.0011 http://doi.org/10.3171/jns.1998.88.1.0011
SONG Q Q , RUIZ J , XING F , et al . Single-cell sequencing reveals the landscape of the human brain metastatic microenvironment [J ] . Commun Biol , 2023 , 6 ( 1 ): 760 . DOI: 10.1038/s42003-023-05124-2 http://doi.org/10.1038/s42003-023-05124-2
CHI Y D , REMSIK J , KISELIOVAS V , et al . Cancer cells deploy lipocalin-2 to collect limiting iron in leptomeningeal metastasis [J ] . Science , 2020 , 369 ( 6501 ): 276 - 282 . DOI: 10.1126/science.aaz2193 http://doi.org/10.1126/science.aaz2193
BOIRE A , ZOU Y L , SHIEH J , et al . Complement component 3 adapts the cerebrospinal fluid for leptomeningeal metastasis [J ] . Cell , 2017 , 168 ( 6 ): 1101 - 1113 .e13. DOI: S0092-8674(17)30240-4 http://doi.org/S0092-8674(17)30240-4
NOLAN C , DEANGELIS L M . Overview of metastatic disease of the central nervous system [J ] . Handb Clin Neurol , 2018 , 149 : 3 - 23 . DOI: B978-0-12-811161-1.00001-3 http://doi.org/B978-0-12-811161-1.00001-3
MANSOORI B , MOHAMMADI A , DITZEL H J , et al . HMGA2 as a critical regulator in cancer development [J ] . Genes (Basel) , 2021 , 12 ( 2 ): 269 .
HASHEMI M , RASHIDI M , HUSHMANDI K , et al . HMGA2 regulation by miRNAs in cancer: affecting cancer hallmarks and therapy response [J ] . Pharmacol Res , 2023 , 190 : 106732 .
XU J X , FANG X J , LONG L Y , et al . HMGA2 promotes breast cancer metastasis by modulating Hippo-YAP signaling pathway [J ] . Cancer Biol Ther , 2021 , 22 ( 1 ): 5 - 11 . DOI: 10.1080/15384047.2020.1832429 http://doi.org/10.1080/15384047.2020.1832429
ZHANG S , ZHANG H , YU L . HMGA2 promotes glioma invasion and poor prognosis via a long-range chromatin interaction [J ] . Cancer Med , 2018 , 7 ( 7 ): 3226 - 3239 .
KAUR H , ALI S Z , HUEY L , et al . The transcriptional modulator HMGA2 promotes stemness and tumorigenicity in glioblastoma [J ] . Cancer Lett , 2016 , 377 ( 1 ): 55 - 64 . DOI: 10.1016/j.canlet.2016.04.020 http://doi.org/10.1016/j.canlet.2016.04.020
VAN DER MAATEN L , HINTON G . Visualizing data using t -SNE [J ] . J Mach Learn Res , 2008 , 9 : 2579 - 605 .
KOZOVSKA Z , GABRISOVA V , KUCEROVA L . Malignant melanoma: diagnosis, treatment and cancer stem cells [J ] . Neoplasma , 2016 , 63 ( 4 ): 510 - 517 . DOI: 10.4149/neo_2016_403 http://doi.org/10.4149/neo_2016_403
YDE S S , SJOEGREN P , HEJE M , et al . Mucosal melanoma: a literature review [J ] . Curr Oncol Rep , 2018 , 20 ( 3 ): 1 - 10 .
韩如雪 , 梁翔 , 马旭辉 , 等 . 口腔黏膜恶性黑色素瘤肺转移特征及预后分析 [J ] . 中国癌症杂志 , 2022 , 32 ( 12 ): 1184 - 1189 . DOI: 10.19401/j.cnki.1007-3639.2022.12.006 http://doi.org/10.19401/j.cnki.1007-3639.2022.12.006
HAN R X , LIANG X , MA X H , et al . Characteristics and prognostic analysis of oral mucosal melanoma with lung metastasis [J ] . China Oncol , 2022 , 32 ( 12 ): 1184 - 1189 .
STEININGER J , GELLRICH F F , ENGELLANDT K , et al . Leptomeningeal metastases in melanoma patients: an update on and future perspectives for diagnosis and treatment [J ] . Int J Mol Sci , 2023 , 24 ( 14 ): 11443 .
PAPE E , DESMEDT E , ZAIRI F , et al . Leptomeningeal metastasis in melanoma: a prospective clinical study of nine patients [J ] . In Vivo , 2012 , 26 ( 6 ): 1079 - 1086 .
THAKKAR J P , KUMTHEKAR P , DIXIT K S , et al . Leptomeningeal metastasis from solid tumors [J ] . J Neurol Sci , 2020 , 411 : 116706 .
RAIZER J J , HWU W J , PANAGEAS K S , et al . Brain and leptomeningeal metastases from cutaneous melanoma: survival outcomes based on clinical features [J ] . Neuro Oncol , 2008 , 10 ( 2 ): 199 - 207 .
XIONG L , TANG Y , TANG J , et al . Downregulation of lncRNA HOTTIP suppresses the proliferation, migration, and invasion of oral tongue squamous cell carcinoma by regulation of HMGA2-mediated Wnt/β-catenin pathway [J ] . Cancer Biother Radiopharm , 2020 , 35 ( 9 ): 720 - 730 .
LIU C M , ZHENG S F , WANG Z Y , et al . KRAS-G12D mutation drives immune suppression and the primary resistance of anti-PD-1/PD-L1 immunotherapy in non-small cell lung cancer [J ] . Cancer Commun (Lond) , 2022 , 42 ( 9 ): 828 - 847 .
WANG X , WANG J , ZHAO J H , et al . HMGA2 facilitates colorectal cancer progression via STAT3-mediated tumor-associated macrophage recruitment [J ] . Theranostics , 2022 , 12 ( 2 ): 963 - 975 . DOI: 10.7150/thno.65411 http://doi.org/10.7150/thno.65411
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