Hypoxia-inducible factor-2α promotes chemoresistance in glioblastoma cells <italic style="font-style: italic">via</italic> transforming growth factor-β2

ZHAO Lu; WANG Pan; GONG Sheng; LIAO Bin; WU Nan

doi:10.16016/j.2097-0927.202511068

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Hypoxia-inducible factor-2α promotes chemoresistance in glioblastoma cells via transforming growth factor-β2

Neuroscience | 更新时间：2026-03-30

- Hypoxia-inducible factor-2α promotes chemoresistance in glioblastoma cells via transforming growth factor-β2
- Journal of Army Medical University Vol. 48, Issue 6, Pages: 701-708(2026)
- 作者机构：
  
  1.重庆医科大学，重庆
  2.重庆市人民医院神经外科，重庆市脑胶质瘤精准医学研究中心，重庆
- 作者简介：
  
  WU Nan, E-mail：wunan@cqu.edu.cn
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(82473430);the Major Joint Medical Research Project of Chongqing Science and Technology Commission and Chongqing Health Commission(2023DBXM004);the Joint Youth Medical Research Project of Chongqing Science and Technology Commission and Chongqing Health Commission Chongqing(2023QNXM043)
- DOI：10.16016/j.2097-0927.202511068
  CLC： R739.41;R730.53;R329.2
- Received：13 November 2025，
  
  Revised：2026-01-28，
  
  Published：30 March 2026
- 稿件说明：
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赵璐，汪攀，龚胜，等.缺氧诱导因子-2α经转化生长因子-β2促进胶质母细胞瘤细胞化疗抵抗［J］.陆军军医大学学报，2026，48（06）：701-708.DOI：10.16016/j.2097-0927.202511068.

ZHAO L，WANG P，GONG S，et al.Hypoxia-inducible factor-2α promotes chemoresistance in glioblastoma cells via transforming growth factor-β2［J］.J Army Med Univ，2026，48（06）：701-708.DOI：10.16016/j.2097-0927.202511068.
赵璐，汪攀，龚胜，等.缺氧诱导因子-2α经转化生长因子-β2促进胶质母细胞瘤细胞化疗抵抗［J］.陆军军医大学学报，2026，48（06）：701-708.DOI：10.16016/j.2097-0927.202511068. DOI：

ZHAO L，WANG P，GONG S，et al.Hypoxia-inducible factor-2α promotes chemoresistance in glioblastoma cells via transforming growth factor-β2［J］.J Army Med Univ，2026，48（06）：701-708.DOI：10.16016/j.2097-0927.202511068. DOI：

摘要

目的

肿瘤微环境中的缺氧是导致胶质母细胞瘤化疗抵抗的关键因素。本研究通过体外研究初步探讨缺氧条件下缺氧诱导因子-2α（hypoxia-inducible factor-2α，HIF-2α）与转化生长因子-β2（transforming growth factor-β2，TGF-β2）促进胶质母细胞瘤细胞化疗抵抗。

方法

常氧-缺氧培养U87、U118细胞（

=3）并予替莫唑胺处理24、48、72 h后经CCK-8检测细胞增殖，培养48 h后流式细胞仪检测细胞凋亡。常氧培养为对照，缺氧培养U87、U118细胞（

=3）4、8、12及24 h后采用RT-qPCR检测HIF-2α、TGF-β2表达，进一步缺氧培养12、24 h及48 h后，Western blot检测HIF-2α、TGF-β2表达。敲除HIF-2α、TGF-β2后，缺氧培养细胞（

=3）并给予替莫唑胺（temozolomide，TMZ）处理24、48、72 h后经CCK-8检测细胞增殖，培养24 h后在部分细胞的培养基中添加TGF-β2（50 ng/mL）继续缺氧培养细胞（

=3）24 h后流式细胞仪检测细胞凋亡。Western blot检测敲除HIF-2α细胞缺氧培养48 h后TGF-β2表达。

结果

CCK-8和流式细胞检测提示，低氧+TMZ组较常氧+TMZ组细胞的存活率增高（

0.05）、凋亡率降低（

0.05）。RT-qPCR检测显示，与常氧组相比，缺氧培养细胞的HIF-2α、TGF-β2表达随着缺氧时间的延长，表达水平逐渐升高（

0.05），Western blot也证实HIF-2α、TGF-β2表达缺氧培养后呈时间依赖性升高。敲除HIF-2α、敲除TGF

-β2表达后的细胞存活率较对照未敲除组降低（

0.05），细胞凋亡率较对照未敲除组升高（

0.05）。敲除HIF-2α细胞继续缺氧培养后提示TGF-β2表达较对照未敲除HIF-2α细胞组水平下降。

结论

HIF-2α经TGF-β2促进胶质母细胞瘤细胞化疗抵抗。

Abstract

Objective

Hypoxia in the tumor microenvironment is a critical factor contributing to chemoresistance in glioblastoma. This study preliminarily investigates the role of hypoxia-inducible factor-2α (HIF-2α) and transforming growth factor-β2 (TGF-β2) in promoting chemotherapy resistance of glioblastoma cells under hypoxic conditions.

Methods

U87 and U118 cells

)

were cultured under normoxic and hypoxic conditions and treated with temozolomide (TMZ) for 24

and 72 h. Cell proliferation was assessed by CCK-8 assay

and apoptosis was detected by flow cytometry after 48 h of culture. Using normoxic culture as control

U87 and U118 cells (

=3) were subjected to hypoxic culture for 4

and 24 h

followed by RT-qPCR to measure HIF-2α and TGF-β2 mRNA expression. After further hypoxic culture for 12

and 48 h

the protein levels of HIF-2α and TGF-β2 were detected by Western blotting. After knockdown of HIF-2α and TGF-β2

the cells

=3) were cultured under hypoxia and treated with TMZ for 24

and 72 h. Then

the cell proliferation was detected by CCK-8 assay. After 24 h of culture

TGF-β2 (50 ng/mL) was added to the culture medium of some cells

and the cells were further cultured under hypoxia for another 24 h

followed by flow cytometry for cell apoptosis. Western blotting was applied to detect TGF-β2 expression in HIF-2α knockdown cells after 48 h of hypoxic culture.

Results

CCK-8 and flow cytometry assays indicated that the hypoxia+TMZ group showed increased cell viability (

0.05) and decreased apoptotic rate compared to the normoxia+TMZ group (

0.05). RT-qPCR revealed that compared with the normoxia group

the expression levels of HIF-2α and TGF-β2 in hypoxia-cultured cells gradually increased with the prolongation of hypoxic time (

0.05)

and Western blotting also confirmed that the protein expression of HIF-2α and TGF-β2 increased in a time-dependent manner after hypoxic culture. Lower cell viability (

0.05) and higher apoptotic rate (

0.05) were observed in the HIF-2α-knockdown and TGF-β2-knockdown cells compared with the control non-knockdown group. Continued hypoxic culture of HIF-2α-knockdown cells indicated decreased TGF-β2 expression compared with the control non-knockdown HIF-2α group.

Conclusion

HIF-2α chemoresistance in glioblastoma cells

via

TGF-β2.

关键词

Keywords

references

SCHAFF L R ， MELLINGHOFF I K . Glioblastoma and other primary brain malignancies in adults： a review ［J］. JAMA ， 2023 ， 329 （ 7 ）： 574 - 587 . DOI： 10.1001/jama.2023.0023 http://dx.doi.org/10.1001/jama.2023.0023 .

CHEN M ， LIU L ， ZENG Y ， et al . An inflammatory gene-related prognostic risk score model for prognosis and immune infiltration in glioblastoma ［J］. Mol Carcinog ， 2024 ， 63 （ 2 ）： 326 - 338 . DOI： 10.1002/mc.23655 http://dx.doi.org/10.1002/mc.23655 .

COLWELL N ， LARION M ， GILES A J ， et al . Hypoxia in the glioblastoma microenvironment： shaping the phenotype of cancer stem-like cells ［J］. Neuro Oncol ， 2017 ， 19 （ 7 ）： 887 - 896 . DOI： 10.1093/neuonc/now258 http://dx.doi.org/10.1093/neuonc/now258 .

WU H B ， GUO C C ， WANG C Y ， et al . Single-cell RNA sequencing reveals tumor heterogeneity， microenvironment， and drug-resistance mechanisms of recurrent glioblastoma ［J］. Cancer Sci ， 2023 ， 114 （ 6 ）： 2609 - 2621 . DOI： 10.1111/cas.15773 http://dx.doi.org/10.1111/cas.15773 .

YUE Q ， WANG Z ， SHEN Y ， et al . Histone H3K9 Lactylation Confers Temozolomide Resistance in Glioblastoma via LUC7L2-Mediated MLH1 Intron Retention ［J］. Adv Sci （Weinh）， 2024 ， 11 （ 19 ）： e2309290 . DOI： 10.1002/advs.202z309290 http://dx.doi.org/10.1002/advs.202z309290 .

DE M M ， DAVIAUD C ， MINNS H E ， et al . Radiation therapy promotes unsaturated fatty acids to maintain survival of glioblastoma ［J］. Cancer Lett ， 2023 ， 570 ： 216329 . DOI： 10.1016/j.canlet.2023.216329 http://dx.doi.org/10.1016/j.canlet.2023.216329 .

VAUPEL P ， MAYER A . Hypoxia in cancer： significance and impact on clinical outcome ［J］. Cancer Metastasis Rev ， 2007 ， 26 （ 2 ）： 225 - 239 . DOI： 10.1007/s10555-007-9055-1 http://dx.doi.org/10.1007/s10555-007-9055-1 .

DAVIS L ， RECKTENWALD M ， HUTT E ， et al . Targeting HIF-2α in the tumor microenvironment： redefining the role of HIF-2α for solid cancer therapy ［J］. Cancers ， 2022 ， 14 （ 5 ）： 1259 . DOI： 10.3390/cancers14051259 http://dx.doi.org/10.3390/cancers14051259 .

KEITH B ， JOHNSON R S ， SIMON M C . HIF1α and HIF2α： sibling rivalry in hypoxic tumour growth and progression ［J］. Nat Rev Cancer ， 2011 ， 12 （ 1 ）： 9 - 22 . DOI： 10.1038/nrc3183 http://dx.doi.org/10.1038/nrc3183 .

APRELIKOVA O ， WOOD M ， TACKETT S ， et al . Role of ETS transcription factors in the hypoxia-inducible factor-2 target gene selection ［J］. Cancer Res ， 2006 ， 66 （ 11 ）： 5641 - 5647 . DOI： 10.1158/0008-5472.CAN-05-3345 http://dx.doi.org/10.1158/0008-5472.CAN-05-3345 .

RENFROW J J ， SOIKE M H ， DEBINSKI W ， et al . Hypoxia-inducible factor 2α： a novel target in gliomas ［J］. Future Med Chem ， 2018 ， 10 （ 18 ）： 2227 - 2236 . DOI： 10.4155/fmc-2018-0163 http://dx.doi.org/10.4155/fmc-2018-0163 .

TZAVLAKI K ， MOUSTAKAS A . TGF-β signaling ［J］. Biomolecules ， 2020 ， 10 （ 3 ）： 487 . DOI： 10.3390/biom10030487 http://dx.doi.org/10.3390/biom10030487 .

WANG M Y ， LIU W J ， WU L Y ， et al . The research progress in transforming growth factor-β2 ［J］. Cells ， 2023 ， 12 （ 23 ）： 2739 . DOI： 10.3390/cells12232739 http://dx.doi.org/10.3390/cells12232739 .

WANG P ， LIAO B ， GONG S ， et al . Temozolomide promotes glioblastoma stemness expression through senescence-associated reprogramming via HIF1α/HIF2α regulation ［J］. Cell Death Dis ， 2025 ， 16 （ 1 ）： 317 . DOI： 10.1038/s41419-025-07617-w http://dx.doi.org/10.1038/s41419-025-07617-w .

YANG S X ， ZHAN Q ， SU D Y ， et al . HIF1α/ATF3 partake in PGK1 K191/K192 succinylation by modulating P4HA1/succinate signaling in glioblastoma ［J］. Neuro Oncol ， 2024 ， 26 （ 8 ）： 1405 - 1420 . DOI： 10.1093/neuonc/noae040 http://dx.doi.org/10.1093/neuonc/noae040 .

邓小玲，张能英，龙开国，等 . 缺氧微环境下HIF-1α与circ-UBE2G1相互调控促进甲状腺癌转移［J］. 陆军军医大学学报， 2025 ， 47 （ 14 ）： 1612 - 1622 . DOI： 10.16016/j.2097-0927.202504040 http://dx.doi.org/10.16016/j.2097-0927.202504040 .

DENG X L ， ZHANG N Y ， LONG K G ， et al . Mutual regulation of HIF-1α and circ-UBE2G1 under hypoxic microenvironment promotes thyroid cancer metastasis ［J］. J Army Med Univ ， 2025 ， 47 （ 14 ）： 1612 - 1622 . DOI： 10.16016/j.2097-0927.202504040 http://dx.doi.org/10.16016/j.2097-0927.202504040 .

LISY K ， PEET D J . Turn me on： regulating HIF transcriptional activity ［J］. Cell Death Differ ， 2008 ， 15 （ 4 ）： 642 - 649 . DOI： 10.1038/sj.cdd.4402315 http://dx.doi.org/10.1038/sj.cdd.4402315 .

WANG P ， ZHAO L ， GONG S ， et al . HIF1α/HIF2α-Sox2/Klf4 promotes the malignant progression of glioblastoma via the EGFR-PI3K/AKT signalling pathway with positive feedback under hypoxia ［J］. Cell Death Dis ， 2021 ， 12 （ 4 ）： 312 . DOI： 10.1038/s41419-021-03598-8 http://dx.doi.org/10.1038/s41419-021-03598-8 .

WANG P ， YAN Q ， LIAO B ， et al . The HIF1α/HIF2α-miR210-3p network regulates glioblastoma cell proliferation， dedifferentiation and chemoresistance through EGF under hypoxic conditions ［J］. Cell Death Dis ， 2020 ， 11 （ 11 ）： 992 . DOI： 10.1038/s41419-020-03150-0 http://dx.doi.org/10.1038/s41419-020-03150-0 .

GOLAN-C I ， CAJA L . The TGF-beta family in glioblastoma ［J］. Int J Mol Sci ， 2024 ， 25 （ 2 ）： 1067 . DOI： 10.3390/ijms25021067 http://dx.doi.org/10.3390/ijms25021067 .

HAU P ， JACHIMCZAK P ， SCHLAIER J ， et al . TGF-β2 signaling in high-grade gliomas ［J］. Curr Pharm Biotechnol ， 2011 ， 12 （ 12 ）： 2150 - 2157 . DOI： 10.2174/13892011179 8808347 http://dx.doi.org/10.2174/138920111798808347 .

ZHAO Y ， WENG Z Y ， ZHOU X ， et al . Mesenchymal stromal cells promote the drug resistance of gastrointestinal stromal tumors by activating the PI3K-AKT pathway via TGF-β2 ［J］. J Transl Med ， 2023 ， 21 （ 1 ）： 219 . DOI： 10.1186/s12967-023-04063-0 http://dx.doi.org/10.1186/s12967-023-04063-0 .

REN D N ， YANG C ， LIU N ， et al . Gene expression profile analysis of U251 glioma cells with shRNA-mediated SOX9 knockdown ［J］. J BUON ， 2018 ， 23 （ 4 ）： 1136 - 1148 .

TANG Y A ， CHEN Y F ， BAO Y ， et al . Hypoxic tumor microenvironment activates GLI2 via HIF-1α and TGF-β2 to promote chemoresistance in colorectal cancer ［J］. Proc Natl Acad Sci USA ， 2018 ， 115 （ 26 ）： E5990 - E5999 . DOI： 10.1073/pnas.1801348115 DOI：10.1073/pnas.1801348115 http://dx.doi.org/10.1073/pnas.1801348115DOI：10.1073/pnas.1801348115 .

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Hypoxia-inducible factor-2α promotes chemoresistance in glioblastoma cells via transforming growth factor-β2

DOI：10.16016/j.2097-0927.202511068

摘要

Abstract

关键词

Keywords

references