INKA2-AS1 promotes migration, invasion and macrophage-related immunosuppression in hepatocellular carcinoma

HE Xiutang; HU Peng; LIU Maonian

doi:10.16016/j.2097-0927.202504005

您当前的位置：

首页 >

文章列表页 >

INKA2-AS1 promotes migration, invasion and macrophage-related immunosuppression in hepatocellular carcinoma

Monographic Reports | 更新时间：2025-09-11

- INKA2-AS1 promotes migration, invasion and macrophage-related immunosuppression in hepatocellular carcinoma
- Journal of Army Medical University Vol. 47, Issue 17, Pages: 2039-2052(2025)
- 作者机构：
  
  1.荆楚理工学院教学质量监控与评估中心，湖北荆门
  2.重庆海吉亚医院肝胆外科，重庆
- 作者简介：
  
  LIU Maonian， E-mail： liumn850614@163.com
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81972760)
- DOI：10.16016/j.2097-0927.202504005
  CLC： R394.3;R730.23;R735.7
- Received：01 April 2025，
  
  Revised：2025-07-26，
  
  Accepted：30 July 2025，
  
  Published：15 September 2025
- 稿件说明：
移动端阅览
何秀堂，胡鹏，刘茂年.INKA2-AS1促进肝细胞癌迁移、侵袭和巨噬细胞相关免疫抑制机制研究［J］.陆军军医大学学报，2025，47（17）：2039-2052.DOI：10.16016/j.2097-0927.202504005.

HE X T，HU P，LIU M N.INKA2-AS1 promotes migration， invasion and macrophage-related immunosuppression in hepatocellular carcinoma［J］.J Army Med Univ，2025，47（17）：2039-2052.DOI：10.16016/j.2097-0927.202504005.
何秀堂，胡鹏，刘茂年.INKA2-AS1促进肝细胞癌迁移、侵袭和巨噬细胞相关免疫抑制机制研究［J］.陆军军医大学学报，2025，47（17）：2039-2052.DOI：10.16016/j.2097-0927.202504005. DOI：

HE X T，HU P，LIU M N.INKA2-AS1 promotes migration， invasion and macrophage-related immunosuppression in hepatocellular carcinoma［J］.J Army Med Univ，2025，47（17）：2039-2052.DOI：10.16016/j.2097-0927.202504005. DOI：

摘要

目的

探究INKA2-AS1促进肝细胞癌（hepatocellular carcinoma，HCC）迁移、侵袭和巨噬细胞相关免疫抑制的机制。

方法

对癌症基因组图谱（the Cancer Genome Atlas Program，TCGA）-HCC数据和健康组织基因表达数据（Genotype-Tissue Expression，GTEx）数据库中374例HCC组织和160例正常肝组织中的基因表达水平进行差异分析。通过生存分析研究INKA2-AS1与HCC患者预后的关系，并对INKA2-AS1进行Cox回归分析。通过肿瘤免疫估计资源数据库分析INKA2-AS1与HCC组织免疫浸润的关系。于2019年1月至2021年1月收集在重庆海吉亚医院肝胆外科接受治疗的90例HCC患者的HCC组织和癌旁正常组织。将HCC细胞Hepa1-6分为NC、INKA2-AS1、siNC和siINKA2-AS1组。通过转染来过表达和沉默INKA2-AS1。分别通过CCK-8、EdU、TUNEL染色、划痕愈合和Transwell实验检测各组细胞的增殖、凋亡、迁移、侵袭情况。将各组Hepa 1-6细胞分别与巨噬细胞共培养分析其对巨噬细胞的影响。将20只4周龄SPF级裸鼠（体质量15~19 g，雌雄各半）通过随机数字表法分为4组（

=5）：NC、INKA2-AS1、siNC和siINKA2-AS1组，分别通过皮下注射2×10

个INKA2-AS1沉默和过表达的Hepa 1-6细胞构建荷瘤裸鼠模型，检测INKA2-AS1对肿瘤体积、质量的影响。通过Western blot检测肿瘤组织中IL-10和FAP蛋白表达水平来分析肿瘤相关巨噬细胞水平。

结果

HCC组织中INKA2-AS1的水平显著高于正常组织（

0.001）。INKA2-AS1与HCC的组织学分级、病理分期有关（

0.05）。NKA2-AS1高表达的HCC患者的生存率（

0.001）、疾病特异性生存期（

0.036）和无进展间期（

0.024）显著低于INKA2-AS1低表达患者。Cox回归分析显示INKA2-AS1是HCC患者OS的独立预测因素（

=0.005）。INKA2-AS1的表达与T辅助细胞、巨噬细胞等免疫细胞相关。HCC组织中INKA2-AS1水平显著高于癌旁正常组织（

0.05）。siINKA2-AS1组的增殖、迁移、侵袭能力低于siNC组（

0.05），凋亡率显著高于siNC组（

0.05），抑制与其共培养的巨噬细胞中IL-10和FAP mRNA和蛋白的表达（

0.05）。INKA2-AS1组的增殖、迁移、侵袭能力高于NC组（

0.05），凋亡率显著低于NC组（

0.05），促进与其共培养的巨噬细胞中IL-10和FAP mRNA和蛋白的表达（

0.01）。siINKA2-AS1组的肿瘤体积和质量显著低于siNC组（

0.05），而

INKA2-AS1组的肿瘤体积（

0.05）和质量显著高于NC组（

0.01）。siINKA2-AS1组肿瘤组织中IL-10和FAP蛋白水平显著低于siNC组（

0.05），INKA2-AS1组肿瘤组织中IL-10和FAP蛋白水平显著高于NC组（

0.05）。

结论

INKA2-AS1升高与HCC患者的预后不良有关，INKA2-AS1促进HCC细胞的增殖和转移，并诱导肿瘤相关巨噬细胞的转化。

Abstract

Objective‍　‍To investigate the mechanism by which INKA2-AS1 promotes hepatocellular carcinoma(HCC) migration

invasion

and macrophage-related immunosuppression. Methods‍　‍Based on the Cancer Genome Atlas Program (TCGA)-HCC data and Genotype-Tissue Expression (GTEx) databases

differentially expressed genes were analyzed in 374 HCC tissues and 160 normal liver tissues. Survival analysis was used to investigate the relationship between INKA2-AS1 and the prognosis of HCC patients. Cox regression analysis was performed on INKA2-AS1. The relationship between INKA2-AS1 and immune infiltration in HCC tissues was analyzed using the Tumor Immune Estimation Resource Database. A total of 90 HCC patients were recruited from the Department of Hepatobiliary Surgery

Chongqing Hygeia Hospital from January 2019 to January 2021. The HCC tissues and adjacent normal tissues were collected. Hepa 1-6 liver cancer cells were grouped into: normal control (NC)

INKA2-AS1

siNC and siINKA2-AS1. INKA2-AS1 was overexpressed and silenced by transfection. The proliferation

migration and invasion of cells in each group were detected by CCK-8

EdU

TUNEL

wound healing and Transwell assays. Each group of Hepa 1-6 cells were co-cultured with macrophages to analyze their effects on macrophages. Twenty nude mice (SPF grade

4 weeks old

15~19 g

half male and half female) were randomly divided into 4 groups (

=5): NC

INKA2-AS1

siNC and siINKA2-AS1. Tumor-bearing nude mouse models were established by subcutaneous injection of INKA2-AS1 silenced and overexpressed Hepa1-6 cells. The effect of INKA2-AS1 on tumor volume and mass were detected. M

acrophage immunity was analyzed by Western blot analysis of IL-10 and FAP protein levels in tumor tissues. Results‍　‍The level of INKA2-AS1 in HCC tissues was significantly higher than that in normal tissues(

0.05). INKA2-AS1 was associated with the histological grade

pathological stage of HCC (

0.05). The survival rate (

0.05)

disease-specific survival (

0.036) and progression-free interval (

0.024) of HCC patients with high INKA2-AS1 expression were significantly lower than those of patients with low INKA2-AS1 expression. The level of INKA2-AS1 in HCC tissues was significantly higher than that in adjacent normal tissues (

0.05). Cox regression analysis showed that INKA2-AS1 was an independent predictor of OS in HCC patients (

=0.005). INKA2-AS1 expression was associated with immune cells such as T cells and macrophages. The proliferation

migration

and invasion abilities of the siINKA2-AS1 group were lower than those of the siNC group (

0.05)

the apoptotic rate was higher than that of the siNC group (

0.05)

and inhibited the expression of IL-10 and FAP at mRNA and protein levels in co-cultured macrophages (

0.05). The proliferation

migration and invasion abilities of the INKA2-AS1 group were higher than those of the NC group (

0.05)

the apoptotic rate was lower than that of the NC group (

0.05)

and promoted the expression of IL-10 and FAP at mRNA and protein levels in co-cultured macrophages (

0.05). The tumor volume and mass of the siINKA2-AS1 group were lower than those of the siNC group (

0.05)

while the tumor volume and mass of the INKA2-AS1 group were higher than those of the NC group (

0.01). The levels of IL-10 and FAP proteins in tumor tissues of the siINKA2-AS1 group were lower than those of the s

iNC group (

0.05)

while the levels of IL-10 and FAP proteins were increased (

0.05). Conclusion‍　‍INKA2-AS1 increment is related to poor prognosis of HCC patients

and the molecule promotes the proliferation and metastasis of HCC cells and induces the transformation of tumor-associated macrophages.

关键词

Keywords

references

BRAY F ， LAVERSANNE M ， SUNG H ， et al . Global cancer statistics 2022： GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries ［J］. CA Cancer J Clin ， 2024 ， 74 （ 3 ）： 229 - 263 . DOI： 10.3322/caac.21834 http://dx.doi.org/10.3322/caac.21834 .

SANKAR K ， GONG J ， OSIPOV A ， et al . Recent advances in the management of hepatocellular carcinoma ［J］. Clin Mol Hepatol ， 2023 ， 30 （ 1 ）： 1 - 15 . DOI： 10.3350/cmh.2023.0125 http://dx.doi.org/10.3350/cmh.2023.0125 .

DE MARTINO M ， RATHMELL J C ， GALLUZZI L ， et al . Cancer cell metabolism and antitumour immunity ［J］. Nat Rev Immunol ， 2024 ， 24 （ 9 ）： 654 - 669 . DOI： 10.1038/s41577-024-01026-4 http://dx.doi.org/10.1038/s41577-024-01026-4 .

CAI N ， CHENG K ， MA Y ， et al . Targeting MMP9 in CTNNB1 mutant hepatocellular carcinoma restores CD8 + T cell-mediated antitumour immunity and improves anti-PD-1 efficacy ［J］. Gut ， 2024 ， 73 （ 6 ）： 985 - 999 . DOI： 10.1136/gutjnl-2023-331342 http://dx.doi.org/10.1136/gutjnl-2023-331342 .

XIAO J ， SONG Y X ， GAO R ， et al . Changes of immune microenvironment in head and neck squamous cell carcinoma in 3D-4-culture compared to 2D-4-culture ［J］. J Transl Med ， 2023 ， 21 （ 1 ）： 771 . DOI： 10.1186/s12967-023-04650-1 http://dx.doi.org/10.1186/s12967-023-04650-1 .

ZHANG Y G . LncRNA-encoded peptides in cancer ［J］. J Hematol Oncol ， 2024 ， 17 （ 1 ）： 66 . DOI： 10.1186/s13045-024-01591-0 http://dx.doi.org/10.1186/s13045-024-01591-0 .

ZONG K ， LIN C F ， LUO K ， et al . Ferroptosis-related lncRNA NRAV affects the prognosis of hepatocellular carcinoma via the miR-375-3P/SLC7A11 axis ［J］. BMC Cancer ， 2024 ， 24 （ 1 ）： 496 . DOI： 10.1186/s12885-024-12265-y http://dx.doi.org/10.1186/s12885-024-12265-y .

LI X Y ， LIANG S H ， FEI M M ， et al . LncRNA CRNDE drives the progression of hepatocellular carcinoma by inducing the immunosuppressive niche ［J］. Int J Biol Sci ， 2024 ， 20 （ 2 ）： 718 - 732 . DOI： 10.7150/ijbs.85471 http://dx.doi.org/10.7150/ijbs.85471 .

XU J ， QIAN Y J ， YE M ， et al . Distinct expression profile of lncRNA in endometrial carcinoma ［J］. Oncol Rep ， 2016 ， 36 （ 6 ）： 3405 - 3412 . DOI： 10.3892/or.2016.5173 http://dx.doi.org/10.3892/or.2016.5173 .

CAO W ， CHEN H D ， YU Y W ， et al . Changing profiles of cancer burden worldwide and in China： a secondary analysis of the global cancer statistics 2020 ［J］. Chin Med J ， 2021 ， 134 （ 7 ）： 783 - 791 . DOI： 10.1097/cm9.0000000000001474 http://dx.doi.org/10.1097/cm9.0000000000001474 .

ANWANWAN D ， SINGH S K ， SINGH S ， et al . Challenges in liver cancer and possible treatment approaches ［J］. Biochim Biophys Acta BBA Rev Cancer ， 2020 ， 1873 （ 1 ）： 188314 . DOI： 10.1016/j.bbcan.2019.188314 http://dx.doi.org/10.1016/j.bbcan.2019.188314 .

何凤蝶，王秋童，胡亚男，等 . 中国人群肝癌发病危险因素病例对照研究的meta分析［J］. 现代预防医学， 2022 ， 49 （ 23 ）： 4230 - 4240 . DOI： 10.20043/j.cnki.MPM.202109297 http://dx.doi.org/10.20043/j.cnki.MPM.202109297 .

HE F D ， WANG Q T ， HU Y N ， et al . Meta-analysis of case-control studies on risk factors for liver cancer in the Chinese population ［J］. Mod Prev Med ， 2022 ， 49 （ 23 ）： 4230 - 4240 . DOI： 10.20043/j.cnki.MPM.202109297 http://dx.doi.org/10.20043/j.cnki.MPM.202109297 .

赵海潮，高强 . 2022年度肝癌研究及诊疗新进展［J］. 中国癌症杂志， 2023 ， 33 （ 4 ）： 315 - 326 . DOI： 10.19401/j.cnki.1007-3639.2023.04.002 http://dx.doi.org/10.19401/j.cnki.1007-3639.2023.04.002 .

ZHAO H C ， GAO Q . Progress in research， diagnosis， and treatment of hepatocellular carcinoma in 2022 ［J］. Chin J Oncol ， 2023 ， 33 （ 4 ）： 315 - 326 . DOI： 10.19401/j.cnki.1007-3639.2023.04.002 http://dx.doi.org/10.19401/j.cnki.1007-3639.2023.04.002 .

WANG F ， HU Y H ， WANG H D ， et al . LncRNA FTO-IT1 promotes glycolysis and progression of hepatocellular carcinoma through modulating FTO-mediated N6-methyladenosine modification on GLUT1 and PKM2 ［J］. J Exp Clin Cancer Res ， 2023 ， 42 （ 1 ）： 267 . DOI： 10.1186/s13046-023-02847-2 http://dx.doi.org/10.1186/s13046-023-02847-2 .

ZHAO J J ， CHEN P J ， TAN C ， et al . LncRNA LINC00667 gets involved in clear cell renal cell carcinoma development and chemoresistance by regulating the miR-143-3p/ZEB1 axis ［J］. Aging ， 2023 ， 15 （ 19 ）： 10057 - 10071 . DOI： 10.18632/aging.205029 http://dx.doi.org/10.18632/aging.205029 .

FERREIRA R S ， ASSIS R I F ， RACCA F ， et al . Analyzes in silico indicate the lncRNAs MIR31HG and LINC00939 as possible epigenetic inhibitors of the osteogenic differentiation in PDLCs ［J］. Genes ， 2023 ， 14 （ 8 ）： 1649 . DOI： 10.3390/genes14081649 http://dx.doi.org/10.3390/genes14081649 .

邓钰梅，陈俊霞 . lncRNA ENST 933在乳腺癌中的作用及其机制研究［J］. 陆军军医大学学报， 2024 ， 46 （ 14 ）： 1676 - 1694 . DOI： 10.16016/j.2097-0927.202401022 http://dx.doi.org/10.16016/j.2097-0927.202401022 .

DENG Y M ， CHEN J X . Role of long non-coding RNA ENST 933 in breast cancer and its underlying mechanism ［J］. J Army Med Univ ， 2024 ， 46 （ 14 ）： 1676 - 1694 . DOI： 10.16016/j.2097-0927.202401022 http://dx.doi.org/10.16016/j.2097-0927.202401022 .

ZHANG Q L ， ZHENG L M ， BAI Y K ， et al . ITPR1-AS1 promotes small cell lung cancer metastasis by facilitating P21 splicing and stabilizing DDX3X to activate the cRaf-MEK-ERK cascade ［J］. Cancer Lett ， 2023 ， 577 ： 216426 . DOI： 10.1016/j.canlet.2023.216426 http://dx.doi.org/10.1016/j.canlet.2023.216426 .

QI F ， DU X J ， ZHAO Z Y ， et al . Tumor mutation burden-associated LINC00638/miR-4732-3p/ULBP1 axis promotes immune escape via PD-L1 in hepatocellular carcinoma ［J］. Front Oncol ， 2021 ， 11 ： 729340 . DOI： 10.3389/fonc.2021.729340 http://dx.doi.org/10.3389/fonc.2021.729340 .

SINGH A ， MAYENGBAM S S ， YADUVANSHI H ， et al . Obesity programs macrophages to support cancer progression ［J］. Cancer Res ， 2022 ， 82 （ 23 ）： 4303 - 4312 . DOI： 10.1158/0008-5472.can-22-1257 http://dx.doi.org/10.1158/0008-5472.can-22-1257 .

VITALE I ， MANIC G ， COUSSENS L M ， et al . Macrophages and metabolism in the tumor microenvironment ［J］. Cell Metab ， 2019 ， 30 （ 1 ）： 36 - 50 . DOI： 10.1016/j.cmet.2019.06.001 http://dx.doi.org/10.1016/j.cmet.2019.06.001 .

HELM O ， HELD-FEINDT J ， GRAGE-GRIEBENOW E ， et al . Tumor-associated macrophages exhibit pro- and anti-inflammatory properties by which they impact on pancreatic tumorigenesis ［J］. Int J Cancer ， 2014 ， 135 （ 4 ）： 843 - 861 . DOI： 10.1002/ijc.28736 http://dx.doi.org/10.1002/ijc.28736 .

OSHI M ， TOKUMARU Y ， ASAOKA M ， et al . M1 macrophage and M1/M2 ratio defined by transcriptomic signatures resemble only part of their conventional clinical characteristics in breast cancer ［J］. Sci Rep ， 2020 ， 10 ： 16554 . DOI： 10.1038/s41598-020-73624-w http://dx.doi.org/10.1038/s41598-020-73624-w .

WU K Y ， LIN K J ， LI X Y ， et al . Redefining tumor-associated macrophage subpopulations and functions in the tumor microenvironment ［J］. Front Immunol ， 2020 ， 11 ： 1731 . DOI： 10.3389/fimmu.2020.01731 http://dx.doi.org/10.3389/fimmu.2020.01731 .

WENES M ， SHANG M ， DI MATTEO M ， et al . Macrophage metabolism controls tumor blood vessel morphogenesis and metastasis ［J］. Cell Metab ， 2016 ， 24 （ 5 ）： 701 - 715 . DOI： 10.1016/j.cmet.2016.09.008 http://dx.doi.org/10.1016/j.cmet.2016.09.008 .

WU Y Y ， WU C ， SHI T C ， et al . FAP expression in adipose tissue macrophages promotes obesity and metabolic inflammation ［J］. Proc Natl Acad Sci U S A ， 2023 ， 120 （ 51 ）： 5120 - 5132 . DOI： 10.1073/pnas.2303075120 http://dx.doi.org/10.1073/pnas.2303075120 .

ZHAO J L ， LIN E ， BAI Z R ， et al . Cancer-associated fibroblasts induce sorafenib resistance of hepatocellular carcinoma cells through CXCL12/FOLR1 ［J］. BMC Cancer ， 2023 ， 23 （ 1 ）： 1198 . DOI： 10.1186/s12885-023-11613-8 http://dx.doi.org/10.1186/s12885-023-11613-8 .

CHONG L J ， JIANG Y W ， WANG D X ， et al . Targeting and repolarizing M2-like tumor-associated macrophage-mediated MR imaging and tumor immunotherapy by biomimetic nanoparticles ［J］. J Nanobiotechnol ， 2023 ， 21 （ 1 ）： 401 . DOI： 10.1186/s12951-023-02122-8 http://dx.doi.org/10.1186/s12951-023-02122-8 .

KERZEL T ， GIACCA G ， BERETTA S ， et al . In vivo macrophage engineering reshapes the tumor microenvironment leading to eradication of liver metastases ［J］. Cancer Cell ， 2023 ， 41 （ 11 ）： 1892 - 1910.e10 . DOI： 10.1016/j.ccell.2023.09.014 http://dx.doi.org/10.1016/j.ccell.2023.09.014 .

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Emerging advances in clinical diagnosis and treatment of hepatocellular carcinoma: evolution of comprehensive treatment strategies

PD-L1 promotes hepatocellular carcinoma progression by inhibiting PTP1B and activating FAK

Nuclear EGFR cooperates with YY1 to drive CEMIP expression and enhance hepatocellular carcinoma invasion

GPX7 regulates proliferation and senescence of hepatocellular carcinoma cells through the PI3K-AKT pathway

Ferrostatin-1 attenuates inflammatory response to hypoxic lung injury at plateau by inhibiting ferroptosis in lung epithelial cells

Related Author

LI Jianwei

WANG Xingru

LIU Yang

WANG Yuran

YU Ying

TU Li

WANG Zengli

OUYANG Shiqi

Related Institution

Institute of Hepatobiliary Surgery of PLA，First Affiliated Hospital， Army Medical University （Third Military Medical University）

No.953，Xigaze， Tibet Autonomous Region

Department of Biochemistry and Molecular Biology

Class

Grade Majoring in Clinical Medicine， School of Medicine， Anhui University of Science and Technology

⁰