前列腺癌细胞焦亡相关基因预后分析及NLRP1对前列腺癌细胞焦亡的调节作用

Prognostic analysis of genes related to pyroptosis in prostate cancer cells and the regulatory role of NLRP1

摘要

目的 利用基因表达数据库分析前列腺癌（PCa）细胞焦亡相关基因（PRGs）与PCa预后的关系，探究核苷酸结合寡聚化结构域样受体1（NLRP1）在PCa细胞焦亡中的调节机制。方法 从癌症基因组图谱（TCGA）数据库、基因表达综合数据库（GEO）获取PCa、癌旁正常组织每千个碱基的转录每百万映射读取的片段（FPKM）数据及临床资料。分析PCa与癌旁正常组织差异表达的PRGs，划分亚型并绘制生存曲线。单因素Cox回归分析、最小绝对收缩选择算子（LASSO）回归分析筛选预后相关PRGs，计算风险评分，建立预后风险模型并进行验证。以训练集、验证集的中位风险评分将患者分为高、低风险组，对差异表达的PRGs进行基因本体论（GO）富集和京都基因与基因组百科全书（KEGG）分析。实时荧光定量聚合酶链式反应（qRT-PCR）检测PCa细胞系NLRP1表达水平，诱导DU145、LNCaP细胞焦亡并观察形态变化，蛋白质免疫印迹法（WB）检测焦亡相关分子表达。结果 共获得CHMP4C、CYCS、GPX4、GSDMB、NLRP1、PLCG1 6个具有预后价值的PRGs，患者风险评分与复发风险呈正相关，与无进展生存期呈负相关（P＜0.001）。训练集1、3、5年AUC分别为0.769（95%CI:0.652～0.878）、0.804（95%CI:0.736～0.882）、0.772（95%CI:0.631～0.905），验证集分别为0.731（95%CI:0.647～0.826）、0.753（95%CI:0.674～0.818）、0.763（95%CI:0.626～0.849）。训练集和验证集中高、低风险组的6个PRGs表达水平均存在差异（P＜0.05）。Cox回归分析显示T分期、前列腺特异性抗原（PSA）、Gleason分级、风险评分为PCa预后的独立预测因素（P＜0.05），年龄、T分期、Gleason分级患者其风险评分差异均有统计学意义（P＜0.05）。NLRP1在PCa细胞系中低表达，参与DU145、LNCaP细胞焦亡调节。结论 基于PRGs构建的预后风险模型对PCa患者预后具有一定预测能力，NLRP1可能参与了PCa细胞焦亡的调节过程。

Abstract

Objective To analyze the prognostic value of prostate cancer (PCa) pyroptosis-related genes (PRGs) using gene expression databases and to explore the regulatory mechanism of nucleotide binding oligomerization domain-like receptor containing pyrin domain 1 (NLRP1) in the pyroptosis of PCa cells. Methods Fragments per kilobase of exon model per million reads mapped (FPKM) data and clinical information from PCa and adjacent tissues from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) were obtained. Differentially expressed PRGs between PCa and adjacent tissues, classified subtypes and plotted survival curves were analyzed. Univariate Cox regression analysis, least absolute shrinkage and selection operator (LASSO) regression analysis were conducted to screen prognosis-related PRGs, risk scores were calculated, and a prognostic risk model was constructed and validated. Patients were divided into high and low risk groups based on the median risk scores from the training and validation sets, and gene ontology (GO) enrichment and kyoto encyclopedia of genes and genomes (KEGG) analysis were conducted on differentially expressed PRGs. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression level of NLRP1 in PCa cell lines, and pyroptosis was induced in DU145 and LNCaP cells while morphological changes were observed. Western blot (WB) was performed to detect the expression of pyroptosis-related molecules. Results A total of 6 prognostic-related PRGs were obtained, including CHMP4C, CYCS, GPX4, GSDMB, NLRP1, and PLCG1. The risk score was positively correlated with the risk of recurrence but negatively correlated with the progression-free survival (P<0.001). The area under the receiver operating characteristic curves (AUCs) for the training set at 1, 3, and 5 years were 0.769 (95%CI:0.652-0.878), 0.804 (95%CI:0.736-0.882), and 0.772 (95%CI:0.631-0.905), respectively, while those for the validation set were 0.731 (95%CI:0.647-0.826), 0.753 (95%CI:0.674-0.818), and 0.763 (95%CI:0.626-0.849), respectively. Differences in expression levels of the 6 PRGs were observed between the high and low risk groups in both the training and validation sets (P<0.05). Cox regression analysis showed that T stage, prostate specific antigen (PSA), Gleason grade, and risk score were independent predictors of PCa prognosis (P<0.05). Differences in risk scores were observed among patients of different ages, T stages, and Gleason grades (P<0.05). NLRP1 was found to be lowly expressed in PCa cell lines and was involved in the regulation of pyroptosis in DU145 and LNCaP cells. Conclusion The prognostic risk model constructed based on PRGs has a certain predictability for the prognosis of PCa patients, and NLRP1 may be involved in the regulation of pyroptosis in PCa cells.

关键词

前列腺癌 / 细胞焦亡 / 核苷酸结合寡聚化结构域样受体1 / 癌症基因组图谱数据库

Key words

prostate cancer / pyroptosis / nucleotide binding oligomerization domain-like receptor containing pyrin domain 1 / The Cancer Genome Atlas

参考文献

[1] EPSTEIN JI, KRYVENKO ON. A comparison of genitourinary society pathology and international society of urological pathology prostate cancer guidelines[J]. Eur Urol, 2021, 79(1): 3-5.

[2] LIU Z, GUO H, ZHU Y, et al. TP53 alterations of hormone-naïve prostate cancer in the Chinese population[J]. Prostate Cancer Prostatic Dis, 2021, 24(2): 482-491.

[3] 康海, 刘晓颖, 赵洁, 等. 基于TCGA数据库分析lncRNA SNHG25在前列腺癌中的表达及其意义[J]. 现代泌尿外科杂志, 2024, 29(3): 224-231.

[4] DEVOS G, DE VLIES W, DE MEERLEER G, et al. Neoadjuvant hormonal therapy before radical prostatectomy in high-risk prostate cancer[J]. Nat Rev Urol, 2021, 18(12): 739-762.

[5] PANEBIANCO V, VILLEIRS G, WEINREB JC, et al. Prostate magnetic resonance imaging for local recurrence reporting (PI-RR): international consensus-based guidelines on multiparametric magnetic resonance imaging for prostate cancer recurrence after radiation therapy and radical prostatectomy[J]. Eur Urol Oncol, 2021, 4(6): 868-876.

[6] DANACIOGLU YO, KESER F, EFILOGˇLU Ö, et al. The efficiency of prostate-specific antigen density measurement using three different methods on the prediction of biochemical recurrence[J]. Aging Male, 2021, 24(1): 15-23.

[7] WEI X, XIE F, ZHOU X, et al. Role of pyroptosis in inflammation and cancer[J]. Cell Mol Immunol, 2022, 19(9): 971-992.

[8] RODRÍGUEZ-ANTONIO I, LÓPEZ-SÁNCHEZ GN, URIBE M, et al. Role of the inflammasome, gasdermin D, and pyroptosis in non-alcoholic fatty liver disease[J]. J Gastroenterol Hepatol, 2021, 36(10): 2720-2727.

[9] 李春桃, 唐苏婷, 徐云柯, 等. miR-16-5p靶向NLRP1抑制缺氧复氧诱导的BRL-3A细胞焦亡[J]. 中国比较医学杂志, 2021, 31(10): 68-75.

[10] CHEN T, WANG Z, ZHONG J, et al. Secoisolariciresinol diglucoside induces pyroptosis by activating caspase-1 to cleave GSDMD in colorectal cancer cells[J]. Drug Dev Res, 2022, 83(5): 1152-1166.

[11] DEVANT P, KAGAN JC. Molecular mechanisms of gasdermin D pore-forming activity[J]. Nat Immunol, 2023, 24(7): 1064-1075.

[12] JIA Y, WANG X, DENG Y, et al. Pyroptosis provides new strategies for the treatment of cancer[J]. J Cancer, 2023, 14(1): 140-151.

[13] HU D, CAO Q, TONG M, et al. A novel defined risk signature based on pyroptosis-related genes can predict the prognosis of prostate cancer[J]. BMC Med Genomics, 2022, 15(1): 24-40.

[14] GANDAGLIA G, LENI R, BRAY F, et al. Epidemiology and prevention of prostate cancer[J]. Eur Urol Oncol, 2021, 4(6): 877-892.

[15] TSAURI I, BLAHETA RA, DOTZAUER R, et al. Focal therapy for primary tumor and metastases in de novo or recurrent oligometastatic prostate cancer: current standing and future perspectives[J]. World J Urol, 2023, 41(8): 2077-2090.

[16] LEE W, LIM B, KYUNG YS, et al. Impact of positive surgical margin on biochemical recurrence in localized prostate cancer[J]. Prostate Int, 2021, 9(3): 151-156.

[17] ALBISINNI S, DIAMAND R. Understanding biochemical recurrence after radical prostatectomy: trust biology, not a number[J]. Prostate Cancer Prostatic Dis, 2023, 26(4): 637-638.

[18] FERDINANDUS J, FENDLER WP, FAROLFI A, et al. PSMA PET validates higher rates of metastatic disease for European association of urology biochemical recurrence risk groups: an international multicenter study[J]. J Nucl Med, 2022, 63(1): 76-80.

[19] XIANG R, GE Y, SONG W, et al. Pyroptosis patterns characterized by distinct tumor microenvironment infiltration landscapes in gastric cancer[J]. Genes (Basel), 2021, 12(10): 1535-1556.

[20] YE Y, DAI Q, QI H. A novel defined pyroptosis-related gene signature for predicting the prognosis of ovarian cancer[J]. Cell Death Discov, 2021, 7(1): 71-81.

[21] CHEN H, LUO H, WANG J, et al. Identification of a pyroptosis-related prognostic signature in breast cancer[J]. BMC Cancer, 2022, 22(1): 429-444.

[22] PETSALAKI E, ZACHOS G. The abscission checkpoint: a guardian of chromosomal stability[J]. Cells, 2021, 10(12): 3350-3368.

[23] LIU B, GUO S, LI GH, et al. CHMP4C regulates lung squamous carcinogenesis and progression through cell cycle pathway[J]. J Thorac Dis, 2021, 13(8): 4762-4774.

[24] KHAN AA, AL-MAHROUQI N, AL-YAHYAEE A, et al. Deciphering urogenital cancers through proteomic biomarkers: a systematic review and meta-analysis[J]. Cancers (Basel), 2023, 16(1): 22-43.

[25] KAN J, HU Y, GE Y, et al. Declined expressions of vast mitochondria-related genes represented by CYCS and transcription factor ESRRA in skeletal muscle aging[J]. Bioengineered, 2021, 12(1): 3485-3502.

[26] SHEN X, WANG H, WENG C, et al. Caspase 3/GSDME-dependent pyroptosis contributes to chemotherapy drug-induced nephrotoxicity[J]. Cell Death Dis, 2021, 12(2): 186-201.

[27] BAZYLIANSKA V, KALPAGE HA, WAN J, et al. Lysine 53 acetylation of cytochrome c in prostate cancer: warburg metabolism and evasion of apoptosis[J]. Cells, 2021, 10(4): 802-822.

[28] XIE Y, KANG R, KLIONSKY DJ, et al. GPX4 in cell death, autophagy, and disease[J]. Autophagy, 2023, 19(10): 2621-2638.

[29] QU M, WANG Y, QIU Z, et al. Necroptosis, pyroptosis, ferroptosis in sepsis and treatment[J]. Shock, 2022, 57(6): 161-171.

[30] SHAO ZC, ZHU BH, HUANG AF, et al. Docosahexaenoic acid reverses epithelial-mesenchymal transition and drug resistance by impairing the PI3K/AKT/Nrf2/GPX4 signalling pathway in docetaxel-resistant PC3 prostate cancer cells[J]. Folia Biol (Praha), 2022, 68(2): 59-71.

[31] HE H, YI L, ZHANG B, et al. USP24-GSDMB complex promotes bladder cancer proliferation via activation of the STAT3 pathway[J]. Int J Biol Sci, 2021, 17(10): 2417-2429.

[32] GÁMEZ-CHIACHIO M, MOLINA-CRESPO Á, RAMOS-NEBOT C, et al. Gasdermin B over-expression modulates HER2-targeted therapy resistance by inducing protective autophagy through Rab7 activation[J]. J Exp Clin Cancer Res, 2022, 41(1): 285-303.

[33] LI T, YANG Z, LI H, et al. Phospholipase Cγ1 (PLCG1) overexpression is associated with tumor growth and poor survival in IDH wild-type lower-grade gliomas in adult patients[J]. Lab Invest, 2022, 102(2): 143-153.

[34] FENINI G, KARAKAYA T, HENNIG P, et al. NLRP1 inflammasome activation in keratinocytes: increasing evidence of important roles in inflammatory skin diseases and immunity[J]. J Invest Dermatol, 2022, 142(9): 2313-2322.

[35] GRISWOLD AR, HUANG HC, BACHOVCHIN DA. The NLRP1 inflammasome induces pyroptosis in human corneal epithelial cells[J]. Invest Ophthalmol Vis Sci, 2022, 63(3): 2-11.

[36] LIN W, CHEN Y, WU B, et al. Identification of the pyroptosis related prognostic gene signature and the associated regulation axis in lung adenocarcinoma[J]. Cell Death Discov, 2021, 7(1): 161-170.