Zhang Xiyuan, Lu Tiange, Shi Chunlei, Ding Mengfei, Wang Ling, Hu Xinting, Wang Xin
Department of Hematology, Shandong Provincial Hospital, Shandong University, Jinan, Shandong, China.
Department of Hematology, Qingdao Central Hospital, University of Health and Rehabilitation Sciences, Qingdao, China.
Front Immunol. 2025 Jul 16;16:1591615. doi: 10.3389/fimmu.2025.1591615. eCollection 2025.
Response gene to complement 32 (RGC32), a complement activation-inducible factor broadly expressed in normal human tissues, has been implicated in tumorigenesis through its dysregulated expression in various malignancies and its involvement in critical oncogenic processes. Despite its established roles in cancer biology, RGC32 remains uncharacterized in diffuse large B-cell lymphoma (DLBCL). This study provides the first comprehensive investigation of RGC32 expression patterns and functional contributions to DLBCL pathogenesis, elucidating its potential as a novel therapeutic target or prognostic biomarker in this disease.
Immunohistochemical (IHC) staining of RGC32 was performed on specimens from 32 Reactive hyperplasia lymphoid (RHL) patients and 80 DLBCL patients. To evaluate the role of RGC32 in DLBCL, lentivirus vectors either encoding shRGC32 or shControl were transfected into DLBCL cell lines. RNA-sequencing (RNA-seq) analysis was performed between shRGC32 and shControl stably transfected OCI-LY1 cells and functional enrichment analyses used gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG). In order to explored its functions , xenograft models were established by subcutaneously injecting shRGC32 and shControl transfected DLBCL cells into SCID beige mice.
Immunohistochemical analysis revealed RGC32 overexpression in DLBCL tissues contrast with RHL, and was associated advanced Ann Arbor stage (p = 0.043), B symptoms (p = 0.020), and poor progression-free survival (p = 0.015) and overall survival (p = 0.035). Functional studies demonstrated that RGC32 knockdown via shRNA significantly suppressed DLBCL cell proliferation and , with xenograft models showing reduced tumor growth and Ki-67 expression. RNA-seq analysis linked RGC32 depletion to downregulation of cell proliferation and impaired DNA damage repair (DDR) mechanisms. Western blot showed RGC32 knockdown could suppress ATM/ATR/CHK1 pathway and increase the tumor mutational burden (TMB). Furthermore, after inhibition of RGC32, infiltration of CD8+ T cells was increased in DLBCL tumor microenvironment (TME).
This study highlights that RGC32 is a novel molecule in DLBCL progression and might be a potential therapeutic target for DLBCL therapy.
补体32反应基因（RGC32）是一种在正常人体组织中广泛表达的补体激活诱导因子，通过其在各种恶性肿瘤中的表达失调及其参与关键致癌过程，已被认为与肿瘤发生有关。尽管RGC32在癌症生物学中已确立了作用，但在弥漫性大B细胞淋巴瘤（DLBCL）中仍未得到充分研究。本研究首次全面调查了RGC32的表达模式及其对DLBCL发病机制的功能贡献，阐明了其作为该疾病新型治疗靶点或预后生物标志物的潜力。
对32例反应性增生性淋巴组织（RHL）患者和80例DLBCL患者的标本进行RGC32的免疫组织化学（IHC）染色。为了评估RGC32在DLBCL中的作用，将编码shRGC32或shControl的慢病毒载体转染到DLBCL细胞系中。对稳定转染shRGC32和shControl的OCI-LY1细胞进行RNA测序（RNA-seq）分析，并使用基因本体论（GO）和京都基因与基因组百科全书（KEGG）进行功能富集分析。为了探索其功能，通过将转染了shRGC32和shControl的DLBCL细胞皮下注射到SCID米色小鼠中建立异种移植模型。
免疫组织化学分析显示，与RHL相比，DLBCL组织中RGC32过表达，且与Ann Arbor分期晚期（p = 0.043）、B症状（p = 0.020）、无进展生存期差（p = 0.015）和总生存期差（p = 0.035）相关。功能研究表明，通过shRNA敲低RGC32可显著抑制DLBCL细胞增殖，异种移植模型显示肿瘤生长和Ki-反应基因对补体32（RGC32）是一种在正常人体组织中广泛表达的补体激活诱导因子，通过其在各种恶性肿瘤中的表达失调及其参与关键致癌过程，已被认为与肿瘤发生有关。尽管RGC32在癌症生物学中已确立了作用，但在弥漫性大B细胞淋巴瘤（DLBCL）中仍未得到充分研究。本研究首次全面调查了RGC32的表达模式及其对DLBCL发病机制的功能贡献，阐明了其作为该疾病新型治疗靶点或预后生物标志物的潜力。
对32例反应性增生性淋巴组织（RHL）患者和80例DLBCL患者的标本进行RGC32的免疫组织化学（IHC）染色。为了评估RGC32在DLBCL中的作用，将编码shRGC32或shControl的慢病毒载体转染到DLBCL细胞系中。对稳定转染shRGC32和shControl的OCI-LY1细胞进行RNA测序（RNA-seq）分析，并使用基因本体论（GO）和京都基因与基因组百科全书（KEGG）进行功能富集分析。为了探索其功能，通过将转染了shRGC32和shControl的DLBCL细胞皮下注射到SCID米色小鼠中建立异种移植模型。
免疫组织化学分析显示，与RHL相比，DLBCL组织中RGC32过表达，且与Ann Arbor分期晚期（p = 0.043）、B症状（p = 0.020）、无进展生存期差（p = 0.015）和总生存期差（p = 0.035）相关。功能研究表明，通过shRNA敲低RGC32可显著抑制DLBCL细胞增殖，异种移植模型显示肿瘤生长和Ki-67表达降低。RNA-seq分析将RGC32的缺失与细胞增殖的下调和DNA损伤修复（DDR）机制受损联系起来。蛋白质印迹显示，敲低RGC32可抑制ATM/ATR/CHK1途径并增加肿瘤突变负担（TMB）。此外，抑制RGC32后，DLBCL肿瘤微环境（TME）中CD8+T细胞的浸润增加。
本研究强调RGC32是DLBCL进展中的一种新分子，可能是DLBCL治疗的潜在靶点。
