Department of Central Laboratory.
Department of Gastroenterology, The Yancheng School of Clinical Medicine of Nanjing Medical University (Yancheng Third People's Hospital), Yancheng, Jiangsu, China.
Anticancer Drugs. 2023 Mar 1;34(3):422-430. doi: 10.1097/CAD.0000000000001445. Epub 2022 Nov 23.
The purpose of this study was to investigate the effect of chemoresistant cancer-associated fibroblasts (R-CAFs) against cisplatin (DDP) on colorectal cancer (CRC) progression. First, clinical tissue samples of chemoresistant or chemosensitive CRC patients were collected to isolate R-CAFs or chemosensitive CAFs (S-CAFs), respectively. HT29 cells or HUVECs were co-cultured with R-CAFs by transwell device. Then the proliferation and apoptosis of HT29 cells were detected with Cell Counting Kit-8 (CCK-8) and flow cytometry. Transwell assay and tube formation assay was used to detect the migration and angiogenesis of HUVECs. In addition, a colorectal cancer transplantation model was established subcutaneously in nude mice by injecting stably transfected HT29 cells and exosomes from different CAF groups, and then the tumor volume and weight were measured and recorded. Hematoxylin and eosin staining, immunohistochemistry, and terminal deoxynucleotidyl transferase dUTP Nick-End Labeling (TUNEL) staining were performed to characterize the histopathological characteristics and apoptosis level of tumor tissues, respectively. S-CAFs and R-CAFs were isolated successfully. HT29 cell co-culture with R-CAFs significantly affected the proliferation and apoptosis of HT29 cells. Exosomes derived from R-CAFs (R-CAFs-Exo) were delivered to HT29 cells, which could induce viability, suppress apoptosis and accelerate the angiogenesis of CRC. In addition, VEGFA was highly expressed in R-CAFs-Exo, which might indicate that R-CAFs could transmit VEGFA through exosomes. Overexpressed VEGFA in R-CAFs apparently regulates the viability, apoptosis, DDP resistance, and angiogenesis of CRC. In-vivo experiments confirmed that R-CAFs-Exo promoted the progression of CRC and DDP resistance by delivering VEGFA . R-CAFs-derived exosomes promote the viability, apoptosis, DDP resistance, and angiogenesis of CRC by delivering VEGFA .
本研究旨在探讨耐药性癌症相关成纤维细胞（R-CAFs）对顺铂（DDP）抵抗的结直肠癌（CRC）进展的影响。首先，收集耐药性或化疗敏感性 CRC 患者的临床组织样本，分别分离 R-CAFs 或化疗敏感性 CAFs（S-CAFs）。通过 Transwell 装置将 HT29 细胞或 HUVECs 与 R-CAFs 共培养。然后，用细胞计数试剂盒-8（CCK-8）和流式细胞术检测 HT29 细胞的增殖和凋亡。Transwell assay 和管形成 assay 用于检测 HUVECs 的迁移和血管生成。此外，通过注射不同 CAF 组来源的 HT29 细胞及其外泌体，在裸鼠皮下建立结直肠癌移植模型，然后测量和记录肿瘤体积和重量。通过苏木精和伊红染色、免疫组织化学和末端脱氧核苷酸转移酶 dUTP 缺口末端标记（TUNEL）染色分别对肿瘤组织的组织病理学特征和细胞凋亡水平进行特征描述。成功分离 S-CAFs 和 R-CAFs。HT29 细胞与 R-CAFs 共培养显著影响 HT29 细胞的增殖和凋亡。源自 R-CAFs 的外泌体（R-CAFs-Exo）被递送到 HT29 细胞，这可以诱导 HT29 细胞的活力，抑制凋亡并加速 CRC 的血管生成。此外，R-CAFs-Exo 中高度表达 VEGFA，这可能表明 R-CAFs 可以通过外泌体传递 VEGFA。R-CAFs 中过表达的 VEGFA 明显调节 CRC 的活力、凋亡、DDP 耐药性和血管生成。体内实验证实，R-CAFs-Exo 通过传递 VEGFA 促进 CRC 的进展和 DDP 耐药性。R-CAFs 衍生的外泌体通过传递 VEGFA 促进 CRC 的活力、凋亡、DDP 耐药性和血管生成。
