收稿日期: 2025-01-03
录用日期: 2025-03-25
网络出版日期: 2025-06-25
基金资助
江苏省第五期“333工程”培养资金资助项目(BRA2019193)
KIAA1429 inhibits colorectal cancer tumor immunity by regulating PD-L1 and CD8+ T cell
Received date: 2025-01-03
Accepted date: 2025-03-25
Online published: 2025-06-25
目的:研究N6-甲基化腺嘌呤(N6-methyladenosine,m6A)甲基化转移酶KIAA1429在结直肠癌(colorectal cancer,CRC)中对程序性细胞死亡配体1(programmed deathligand-1,PD-L1)表达及CD8+ T细胞浸润的影响。方法:利用在线数据库分析KIAA1429在CRC中的表达对PD-L1表达、CD8+T细胞浸润的影响。收集2020年至2022年南京医科大学附属泰州人民医院连续收治的经病理确诊的CRC患者的癌组织样本及癌旁组织样本,采用免疫组织化学染色法检测,并比较其中KIAA1429、PD-L1水平及CD8+ T细胞浸润情况。在CRC细胞株中,采用定量聚合酶链反应(quantitative polymerase chain reaction,qPCR)及蛋白印迹法分别检测敲减KIAA1429基因后CRC细胞中的PD-L1 mRNA及蛋白表达水平。在CRC同源移植瘤小鼠中敲减KIAA1429基因后,每3 d记录一次肿瘤大小及小鼠体重,利用流式细胞术检测瘤组织中CD8+T细胞浸润水平变化。结果:生物信息分析表明,KIAA1429在CRC中高表达(P<0.05),且与不良预后相关(P=0.028),并与CD8+ T细胞浸润程度呈负相关(P=3.981×10-2)。病理组织免疫组化结果显示,CRC中KIAA1429的表达水平显著高于癌旁组织(P=2.196×10-7),且KIAA1429与PD-L1的表达水平呈正相关(P=1.017×10-7),与CD8+T细胞的浸润程度呈负相关(P=0.021)。在CRC细胞中敲减KIAA1429基因,能够下调PD-L1的mRNA及蛋白表达(P<0.01)。在CRC移植瘤小鼠中敲减KIAA1429基因可以抑制肿瘤生长(P<0.01),提高CD8+T细胞浸润水平(P<0.05)。结论:m6A甲基化转移酶KIAA1429可能通过上调PD-L1表达和减少CD8+ T免疫细胞浸润,从而抑制CRC肿瘤免疫。靶向KIAA1429可能有助于改善CRC患者预后,提高CRC免疫治疗疗效。
杜雅洁 , 王铭飞 , 林茂松 . 结直肠癌中KIAA1429通过上调PD-L1及下调CD8+ T细胞组织浸润而抑制抗肿瘤免疫的研究[J]. 诊断学理论与实践, 2025 , 24(03) : 301 -311 . DOI: 10.16150/j.1671-2870.2025.03.009
Objective This study aims to investigate the effect of the N6-methyladenosine (m6A) methyltransferase KIAA1429 on programmed death-ligand 1 (PD-L1) expression and CD8+ T cell infiltration in colorectal cancer (CRC). Methods The effects of KIAA1429 expression on PD-L1 expression and CD8+T cell infiltration in CRC were analyzed using online databases. Tissue samples from tumor and adjacent normal regions were collected from patients with pathologically confirmed CRC treated at the Affiliated Taizhou People's Hospital of Nanjing Medical University, admitted and diagnosed between 2020 and 2022. Immunohistochemical (IHC) staining was performed to evaluate and compare the expression levels of KIAA1429 and PD-L1, as well as the degree of CD8+ T cell infiltration. In CRC cell lines, quantitative polymerase chain reaction (qPCR) and western blotting were used to measure PD-L1 mRNA and protein expression levels after KIAA1429 knockdown, respectively. After KIAA1429 knockdown in CRC syngeneic mouse models, tumor size and body weight were recorded every 3 days, and flow cytometry was used to assess changes in CD8+ T cell infiltration degree within the tumor tissue. Results Bioinformatics analysis indicated that KIAA1429 was highly expressed in CRC (P<0.05), associa-ted with poor prognosis (P=0.028), and negatively correlated with CD8+T cell infiltration degree (P=3.981×10-2). IHC results demonstrated that KIAA1429 expression levels in CRC were significantly higher than in adjacent normal tissues (P=2.196×10-7), positively correlated with PD-L1 expression levels (P=1.017×10-7), and negatively correlated with CD8+T cell infiltration degree (P=0.021). KIAA1429 knockdown in CRC cells downregulated both PD-L1 mRNA and protein expression levels (P<0.01). In the CRC syngeneic mouse models, KIAA1429 knockdown inhibited tumor growth (P<0.01) and enhanced CD8+T cell infiltration degree (P<0.05). Conclusions The m6A methyltransferase KIAA1429 may suppress tumor immunity in CRC by upregulating PD-L1 expression and reducing CD8+T cell infiltration. Targeting KIAA1429 may help improve the prognosis of CRC patients and enhance the efficacy of CRC immunotherapy.
| [1] | FILHO A M, LAVERSANNE M, FERLAY J, et al. The GLOBOCAN 2022 cancer estimates: data sources, methods, and a snapshot of the cancer burden worldwide[J]. Int J Cancer,2025, 156(7):1336-1346. |
| [2] | 肖毅, 李珂璇. 多中心直肠癌真实世界数据库建设与数据质量控制策略[J]. 中华消化外科杂志 2025, 24(1): 77-81. |
| XIAO Y, LI K X. Multicenter rectal cancer real-world database construction and data quality control strategies[J]. Chin J Dig Surg, 2025, 24(1):77-81. | |
| [3] | 李珂璇, 肖体先, 汪晓东, 等. 中低位直肠癌初诊及新辅助治疗后评估完成度分析:全国多中心真实世界研究[J]. 中华消化外科杂志, 2025, 24(1):113-119. |
| LI K X, XIAO T X, WANG X D, et al. Analysis of completion rate of tumor evaluation at initial assessment and after neoa-djuvant therapy for mid and low rectal cancer : a national multicenter real world study[J]. Chin J Dig Surg, 2025, 24(1):113-119. | |
| [4] | 中华医学会外科学分会腹腔镜与内镜外科学组, 中华医学会外科学分会结直肠外科学组, 中国医师协会外科医师分会结直肠外科专家工作组, 等. 腹腔镜结直肠癌根治术操作指南(2023版)[J]. 中华消化外科杂志, 2024, 23(1):10-22. |
| Laparoscopic & Endoscopic Surgery Group, Branch of Surgery, Chinese Medical Association, Colorectal Surgery Group, Branch of Surgery, Chinese Medical Association, Chinese Society of Colon and Rectal Surgeons, Chinese Medical Doctor Association, et al. Guideline for operative procedure of laparoscopic radical surgery for colorectal cancer (2023 edition)[J]. Chin J Dig Surg, 2024, 23(1): 10-22. | |
| [5] | MORGAN E, ARNOLD M, GINI A, et al. Global burden of colorectal cancer in 2020 and 2040: incidence and morta-lity estimates from GLOBOCAN[J]. Gut, 2023, 72(2): 338-344. |
| [6] | LAZAROFF J, BOLOTIN D. Targeted therapy and immunotherapy in melanoma[J]. Dermatol Clin, 2023, 41(1):65-77. |
| [7] | DANTOING E, PITON N, SALAüN M, et al. Anti-PD1/PD-L1 immunotherapy for non-small cell lung cancer with actionable oncogenic driver mutations[J]. Int J Mol Sci, 2021, 22(12):6288. |
| [8] | JIN M, FANG J, PENG J, et al. PD-1/PD-L1 immune checkpoint blockade in breast cancer: research insights and sensitization strategies[J]. Mol Cancer, 2024, 23(1):266. |
| [9] | MARCUS L, LEMERY S J, KEEGAN P, et al. FDA approval summary: pembrolizumab for the treatment of microsatellite instability-high solid tumors[J]. Clin Cancer Res, 2019, 25(13):3753-3758. |
| [10] | DIAZ L A JR, SHIU K K, KIM T W, et al. Pembrolizumab versus chemotherapy for microsatellite instability-high or mismatch repair-deficient metastatic colorectal cancer (KEYNOTE-177): final analysis of a randomised, open-label, phase 3 study[J]. Lancet Oncol, 2022, 23(5):659-670. |
| [11] | SUN T, WU R, MING L. The role of m6A RNA methylation in cancer[J]. Biomed Pharmacother, 2019,112:108613. |
| [12] | JIANG X, LIU B, NIE Z, et al. The role of m6A modification in the biological functions and diseases[J]. Signal Transduct Target Ther, 2021, 6(1):74. |
| [13] | SHRIWAS O, MOHAPATRA P, MOHANTY S, et al. The impact of m6A RNA modification in therapy resistance of cancer: implication in chemotherapy, radiotherapy, and immunotherapy[J]. Front Oncol, 2021,10:612337. |
| [14] | ZHANG Z, ZHANG C, LUO Y, et al. m6A regulator expression profile predicts the prognosis, benefit of adjuvant chemotherapy, and response to anti-PD-1 immunotherapy in patients with small-cell lung cancer[J]. BMC Med, 2021, 19(1):284. |
| [15] | ZHANG X, LI M J, XIA L, et al. The biological function of m6A methyltransferase KIAA1429 and its role in human disease[J]. PeerJ, 2022,10:e14334. |
| [16] | TANG Z, KANG B, LI C, et al. GEPIA2: an enhanced web server for large-scale expression profiling and interactive analysis[J]. Nucleic Acids Res, 2019, 47(W1):W556-W560. |
| [17] | LI T, FU J, ZENG Z, et al. TIMER2.0 for analysis of tumor-infiltrating immune cells[J]. Nucleic Acids Res, 2020, 48(W1):W509-W514. |
| [18] | CERAMI E, GAO J, DOGRUSOZ U, et al. The cBio cancer genomics portal: an open platform for exploring multidimensional cancer genomics data[J]. Cancer Discov, 2012, 2(5):401-404. |
| [19] | ZHOU Y, ZENG P, LI Y H, et al. SRAMP: prediction of mammalian N6-methyladenosine (m6A) sites based on sequence-derived features[J]. Nucleic Acids Res, 2016, 44(10):e91. |
| [20] | RASKOV H, ORHAN A, CHRISTENSEN J P, et al. Cytotoxic CD8+ T cells in cancer and cancer immunotherapy[J]. Br J Cancer, 2021, 124(2):359-367. |
| [21] | DI PILATO M, GAO Y, SUN Y, et al. Translational studies using the MALT1 Inhibitor (S)-mepazine to induce treg fragility and potentiate immune checkpoint therapy in cancer[J]. J Immunother Precis Oncol, 2023, 6(2):61-73. |
| [22] | ZHOU Y, PEI Z, MAIMAITI A, et al. m6A methyltransfe-rase KIAA1429 acts as an oncogenic factor in colorectal cancer by regulating SIRT1 in an m6A-dependent manner[J]. Cell Death Discov, 2022, 8(1):83. |
| [23] | AI Y, LIU S, LUO H, et al. METTL3 intensifies the progress of oral squamous cell carcinoma via modulating the m6A amount of PRMT5 and PD-L1[J]. J Immunol Res, 2021,2021:6149558. |
| [24] | LIU Z, WANG T, SHE Y, et al. N6-methyladenosine-modified circIGF2BP3 inhibits CD8+ T-cell responses to facilitate tumor immune evasion by promoting the deubi-quitination of PD-L1 in non-small cell lung cancer[J]. Mol Cancer, 2021, 20(1):105. |
| [25] | MA L, LIN Y, SUN S W, et al. KIAA1429 is a potential prognostic marker in colorectal cancer by promoting the proliferation via downregulating WEE1 expression in an m6A-independent manner[J]. Oncogene, 2022, 41(5):692-703. |
| [26] | QIAN J Y, GAO J, SUN X, et al. KIAA1429 acts as an oncogenic factor in breast cancer by regulating CDK1 in an N6-methyladenosine-independent manner[J]. Oncogene, 2019, 38(33):6123-6141. |
| [27] | WAN W, AO X, CHEN Q, et al. METTL3/IGF2BP3 axis inhibits tumor immune surveillance by upregulating N6-methyladenosine modification of PD-L1 mRNA in breast cancer[J]. Mol Cancer, 2022, 21(1):60. |
| [28] | HU Y, ZHAO J, SHEN Y, et al. Predictive value of tumor-infiltrating lymphocytes detected by flow cytometry in colorectal cancer[J]. Int Immunopharmacol, 2022, 113(Pt A):109286. |
| [29] | MALKA D, LIèVRE A, ANDRé T, et al. Immune scores in colorectal cancer: Where are we?[J] Eur J Cancer, 2020,140:105-118. |
| [30] | KARJULA T, ELOMAA H, NISKAKANGAS A, et al. CD3+ and CD8+ T-cell-based immune cell score and PD-(L)1 expression in pulmonary metastases of microsatellite stable colorectal cancer[J]. Cancers (Basel), 2022, 15(1):206. |
| [31] | DOMINGO E, KELLY C, HAY J, et al. Prognostic and predictive value of immunoscore in stage Ⅲ colorectal cancer: pooled analysis of cases from the SCOT and IDEA-HORG studies[J]. J Clin Oncol, 2024, 42(18):2207-2218. |
| [32] | HUANG Y, XIA W, DONG Z, et al. Chemical inhibitors targeting the oncogenic m6A modifying proteins[J]. Acc Chem Res, 2023, 56(21):3010-3022. |
| [33] | CHEN H, PAN Y, ZHOU Q, et al. METTL3 inhibits antitumor immunity by targeting m6A-BHLHE41-CXCL1/CXCR2 axis to promote colorectal cancer[J]. Gastroentero-logy, 2022, 163(4):891-907. |
/
| 〈 |
|
〉 |