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| GRSF1通过AKT信号通路抑制结肠癌EMT及免疫逃逸的机制研究 |
| The mechanism of GRSF1 inhibits EMT and immune Evasion in Colon Cancer through the AKT Signaling Pathway |
| 投稿时间：2024-06-28 修订日期：2024-10-10 |
| DOI： |
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| 中文关键词: GRSF1 AKT 上皮细胞间质转化 免疫浸润 结肠癌 |
| 英文关键词:GRSF1 AKT epithelial-mesenchymal transition immune infiltration colorectal cancer |
| 基金项目:淮安市免疫学重点实验室(项目编号：HAP202002) |
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| 中文摘要: |
| 目的 本次研究的目的是探讨富含鸟嘌呤的RNA序列结合因子1(GRSF1)抑制结肠癌的作用,并研究其潜在的分子机制。方法 TCGA公共数据库分析GRSF1在结肠癌患者中表达及其与患者病理、预后、CD8+T细胞浸润之间的关系。结肠癌组织芯片免疫组织化染色验证GRSF1表达,并分析其表达与患者临床病理、预后及CD8+T细胞浸润之间的关系。通过在人结肠癌细胞株中分别过表达与敲减GRSF1的表达后检测细胞的增殖,侵袭及迁移能力,western blot检测细胞中EMT相关基因表达和PI3K/AKT信号通路的激活,裸鼠皮下建立移植瘤模型,记录肿瘤生长和小鼠生存曲线及转移情况。小鼠结肠癌细胞系MC-38过表达grsf1后建立小鼠结肠移植瘤模型,记录肿瘤生长和小鼠生存曲线及转移情况,流式细胞术检测小鼠肿瘤组织中CD8＋T细胞数量。结果 公共数据库分析显示,结肠癌组织中GRSF1表达较正常组织升高(P<0.05),但患者GRSF1表达越高则预后越好(P<0.05)、CD8+T细胞浸润也越多、淋巴结转移也越少、病理分期也越低(P<0.05)。90例结肠癌患者临床样本验证结果也进一步证实这一结果。人结肠癌细胞过表达GRSF1后细胞增殖、侵袭及迁移能力均显著降低(P<0.05),而敲减GRSF1表达后则细胞增殖、侵袭及迁移能力均显著增强(P<0.05)。western blot显示GRSF1能够调控EMT相关蛋白E-cadherin,N-cadherin,Vimentin的表达,同时GRSF1的表达能够抑制AKT信号通路的激活。过表达GRSF1组小鼠的肿瘤生长曲线较对照组小鼠显著降低(P<0.05),生存曲线较对照组小鼠显著升高(P<0.05),肿瘤组织p-AKT表达也显著低于对照组小鼠(P<0.05)。此外,过表达grsf1组MC-38移植瘤小鼠肿瘤组织中CD8＋T细胞数量较对照组小鼠显著增加(P<0.05)。结论 结肠癌患者肿瘤组织中GRSF1的表达较正常组织升高,GRSF1可能通过调控AKT信号通路抑制结肠癌EMT及免疫逃逸从而发挥抑制结肠癌的作用。 |
| 英文摘要: |
| Objective: The purpose of this study is to investigate the inhibitory effect of Guanine-rich RNA sequence binding factor 1 (GRSF1) on colorectal cancer and to explore its potential molecular mechanisms. Methods: The expression of GRSF1 in colorectal cancer patients and its relationship with patient pathology, prognosis, and CD8+ T cell infiltration were analyzed using the TCGA public database. Immunohistochemical staining of colorectal cancer tissue microarrays was used to verify GRSF1 expression and to analyze its association with clinical pathology, prognosis, and CD8+ T cell infiltration in patients. In human colorectal cancer cell lines, the effects of overexpression and knockdown of GRSF1 on cell proliferation, invasion, and migration were assessed. Western blot was used to detect the expression of EMT-related genes and the activation of the PI3K/AKT signaling pathway. A subcutaneous xenograft tumor model in nude mice was established to record tumor growth, mouse survival curves, and metastasis. In a mouse colorectal cancer cell line MC-38, overexpressing GRSF1 established a mouse colorectal xenograft model to record tumor growth, mouse survival curves, and metastasis. Flow cytometry was used to detect the number of CD8+ T cells in mouse tumor tissues. Results: Analysis of the public database showed that GRSF1 expression in colorectal cancer tissues was higher than in normal tissues (P<0.05). However, higher GRSF1 expression was associated with better prognosis (P<0.05), increased CD8+ T cell infiltration, fewer lymph node metastases, and lower pathological staging (P<0.05). These results were further confirmed by clinical samples from 108 colorectal cancer patients in our hospital. Overexpression of GRSF1 in human colorectal cancer cells significantly reduced cell proliferation, invasion, and migration (P<0.05), whereas knockdown of GRSF1 expression significantly enhanced these abilities (P<0.05). Western blot showed that GRSF1 could regulate the expression of EMT-related proteins E-cadherin, N-cadherin, and Vimentin, and inhibit the activation of the AKT signaling pathway. In mice, the tumor growth curve of the GRSF1 overexpression group was significantly lower than that of the control group (P<0.05), and the survival curve was significantly higher (P<0.05). The expression of p-AKT in tumor tissues was also significantly lower than in the control group (P<0.05). Additionally, the number of CD8+ T cells in the tumor tissues of the GRSF1 overexpression group MC-38 xenograft mice was significantly higher than in the control group (P<0.05). Conclusion: The expression of GRSF1 is higher in tumor tissues of colorectal cancer patients compared to normal tissues. GRSF1 may inhibit colorectal cancer by regulating the AKT signaling pathway to suppress EMT and immune evasion. |
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