Institute of Experimental Medicine, Saint Petersburg, 197376, Russia.
Children's Neurosurgical Department No.7, Almazov Medical Research Centre, 197341, Saint Petersburg, Russia.
Cancer Chemother Pharmacol. 2024 May;93(5):455-469. doi: 10.1007/s00280-023-04622-8. Epub 2024 Jan 27.
Glioblastoma multiforme (GBM) is the most aggressive and fatal malignant primary brain tumor. The enhancement of the survival rate for glioma patients remains limited, even with the utilization of a combined treatment approach involving surgery, radiotherapy, and chemotherapy. This study was designed to assess the expression of IDH1, TP53, EGFR, Ki-67, GFAP, H3K27M, MGMT, VEGF, NOS, CD99, and ATRX in glioblastoma tissue from 11 patients. We investigated the anticancer impact and combined effects of cathelicidin (LL-37), protegrin-1 (PG-1), with chemotherapy-temozolomide (TMZ), doxorubicin (DOX), carboplatin (CB), cisplatin (CPL), and etoposide (ETO) in primary GBM cells. In addition, we examined the effect of LL-37, PG-1 on normal human fibroblasts and in the C6/Wistar rat intracerebral glioma model.
For this study, 11 cases of glioblastoma were evaluated immunohistochemically for IDH1, TP53, EGFR, Ki-67, GFAP, H3K27M, MGMT, VEGF, NOS, CD99, and ATRX. The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was used to study cells viability and to determine cytotoxic effects of LL-37, PG-1 and their combination with chemotherapy in primary GBM cells. Synergism or antagonism was determined using combination index (CI) method. Finally, we established C6 glioblastoma model in Wistar rats to investigate the antitumor activity.
Peptides showed a strong cytotoxic effect on primary GBM cells in the MTT test (IC 2-16 and 1-32 μM) compared to chemotherapy. The dual-drug combinations of LL-37 + DOX, LL-37 + CB (CI 0.46-0.75) and PG-1 + DOX, PG-1 + CB, PG-1 + TMZ (CI 0.11-0.77), demonstrated a synergism in primary GBM cells. In rat C6 intracerebral GBM model, survival of rats in experimental group (66.75 ± 12.6 days) was prolonged compared with that in control cohort (26.2 ± 2.66 days, p = 0.0008). After LL-37 treatment, experimental group rats showed significantly lower tumor volumes (31.00 ± 8.8 mm) and weight (49.4 ± 13.3 mg) compared with control group rats (153.8 ± 43.53 mg, p = 0.038; 82.50 ± 7.60 mm, respectively).
The combination of antimicrobial peptides and chemical drugs enhances the cytotoxicity of chemotherapy and exerts synergistic antitumor effects in primary GBM cells. Moreover, in vivo study provided the first evidence that LL-37 could effectively inhibit brain tumor growth in rat C6 intracerebral GBM model. These results suggested a significant strategy for proposing a promising therapy for the treatment of GBM.
多形性胶质母细胞瘤（GBM）是最具侵袭性和致命性的恶性原发性脑肿瘤。即使采用手术、放疗和化疗相结合的综合治疗方法，也仍然限制了神经胶质瘤患者的生存率的提高。本研究旨在评估 11 例胶质母细胞瘤组织中 IDH1、TP53、EGFR、Ki-67、GFAP、H3K27M、MGMT、VEGF、NOS、CD99 和 ATRX 的表达。我们研究了抗菌肽（LL-37）、防御素-1（PG-1）与替莫唑胺（TMZ）、多柔比星（DOX）、卡铂（CB）、顺铂（CPL）和依托泊苷（ETO）联合化疗对原发性 GBM 细胞的抗癌作用及联合效果。此外，我们还研究了 LL-37、PG-1 对正常人类成纤维细胞的影响，并在 C6/Wistar 大鼠颅内神经胶质瘤模型中进行了研究。
在这项研究中，我们对 11 例胶质母细胞瘤病例进行了免疫组织化学分析，检测 IDH1、TP53、EGFR、Ki-67、GFAP、H3K27M、MGMT、VEGF、NOS、CD99 和 ATRX 的表达。噻唑蓝（MTT）试验用于研究细胞活力，并确定 LL-37、PG-1 及其与化疗联合对原发性 GBM 细胞的细胞毒性作用。采用合并指数（CI）法确定协同作用或拮抗作用。最后，我们在 Wistar 大鼠中建立了 C6 神经胶质瘤模型，以研究抗肿瘤活性。
与化疗相比，肽在 MTT 试验中对原发性 GBM 细胞表现出强烈的细胞毒性作用（IC 2-16 和 1-32 μM）。LL-37+DOX、LL-37+CB（CI 0.46-0.75）和 PG-1+DOX、PG-1+CB、PG-1+TMZ（CI 0.11-0.77）的双重药物组合在原发性 GBM 细胞中表现出协同作用。在大鼠 C6 颅内 GBM 模型中，实验组（66.75±12.6 天）的大鼠存活率较对照组（26.2±2.66 天，p=0.0008）延长。在给予 LL-37 治疗后，实验组大鼠的肿瘤体积（31.00±8.8mm）和重量（49.4±13.3mg）明显低于对照组大鼠（分别为 153.8±43.53mg，p=0.038；82.50±7.60mm）。
抗菌肽与化学药物的联合使用增强了化疗的细胞毒性，并在原发性 GBM 细胞中发挥了协同的抗肿瘤作用。此外，体内研究首次提供了证据表明 LL-37 可有效抑制大鼠 C6 颅内 GBM 模型中的脑肿瘤生长。这些结果为提出一种有希望的治疗胶质母细胞瘤的新策略提供了依据。
