17th Department of Plastic Surgery, Plastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100144, China.
Department of Plastic Surgery, Peking Union Medical College Hospital, Beijing 100730, China.
Chin Med J (Engl). 2021 Sep 13;134(18):2205-2213. doi: 10.1097/CM9.0000000000001780.
Hyperbaric oxygen treatment (HBOT) has been demonstrated to influence the keloid recurrence rate after surgery and to relieve keloid symptoms and other pathological processes in keloids. To explore the mechanism of the effect of HBOT on keloids, tumor immune gene expression and immune cell infiltration were studied in this work.
From February 2021 to April 2021, HBOT was carried out on keloid patients four times before surgery. Keloid tissue samples were collected and divided into an HBOT group (keloid with HBOT before surgery [HK] group, n = 6) and a non-HBOT group (K group, n = 6). Tumor gene expression was analyzed with an Oncomine Immune Response Research Assay kit. Data were mined with R package. The differentially expressed genes between the groups were compared. Hub genes between the groups were determined and verified with Quantitative Real-time PCR. Immune cell infiltration was analyzed based on CIBERSORT deconvolution algorithm analysis of gene expression and verified with immunohistochemistry (IHC).
Inflammatory cell infiltration was reduced in the HK group. There were 178 upregulated genes and 217 downregulated genes. Ten hub genes were identified, including Integrin Subunit Alpha M (ITGAM), interleukin (IL)-4, IL-6, IL-2, Protein Tyrosine Phosphatase Receptor Type C (PTPRC), CD86, transforming growth factor (TGF), CD80, CTLA4, and IL-10. CD80, ITGAM, IL-4, and PTPRC with significantly downregulated expression were identified. IL-10 and IL-2 were upregulated in the HK group but without a significant difference. Infiltration differences of CD8 lymphocyte T cells, CD4 lymphocyte T-activated memory cells, and dendritic resting cells were identified with gene CIBERSORT deconvolution algorithm analysis. Infiltration levels of CD4 lymphocyte T cell in the HK group were significantly higher than those of the K group in IHC verification.
HBOT affected tumor gene expression and immune cell infiltration in keloids. CD4 lymphocyte T cell, especially activated memory CD4+T, might be the key regulatory immune cell, and its related gene expression needs further study.
高压氧治疗（HBOT）已被证明可影响手术后瘢痕疙瘩的复发率，并缓解瘢痕疙瘩的症状和其他病理过程。为了探讨 HBOT 对瘢痕疙瘩影响的机制，本研究探讨了肿瘤免疫基因表达和免疫细胞浸润。
2021 年 2 月至 2021 年 4 月，对 6 例术前接受 HBOT 的瘢痕疙瘩患者（HK 组）和 6 例未接受 HBOT 的瘢痕疙瘩患者（K 组）进行了 4 次 HBOT。采用 Oncomine 免疫反应研究试剂盒分析肿瘤基因表达。采用 R 软件包进行数据挖掘。比较两组间差异表达基因。通过定量实时 PCR 确定和验证两组间的枢纽基因。根据基因表达的 CIBERSORT 去卷积算法分析进行免疫细胞浸润分析，并通过免疫组织化学（IHC）验证。
HK 组炎症细胞浸润减少。有 178 个上调基因和 217 个下调基因。确定了 10 个枢纽基因，包括整合素亚单位 α M（ITGAM）、白细胞介素（IL）-4、IL-6、IL-2、蛋白酪氨酸磷酸酶受体 C 型（PTPRC）、CD86、转化生长因子（TGF）、CD80、细胞毒性 T 淋巴细胞相关抗原 4（CTLA4）和白细胞介素（IL）-10。CD80、ITGAM、IL-4 和 PTPRC 的表达明显下调。HK 组中 IL-10 和 IL-2 上调，但无显著差异。采用 CIBERSORT 去卷积算法分析鉴定 CD8 淋巴细胞 T 细胞、CD4 淋巴细胞 T 激活记忆细胞和树突状静止细胞的浸润差异。IHC 验证发现 HK 组 CD4 淋巴细胞 T 细胞浸润水平明显高于 K 组。
HBOT 影响瘢痕疙瘩的肿瘤基因表达和免疫细胞浸润。CD4 淋巴细胞 T 细胞，尤其是激活的记忆 CD4+T 细胞，可能是关键的调节免疫细胞，其相关基因表达需要进一步研究。
