靶向抑制 Notch1 信号通路对胃癌肿瘤干细胞化疗增敏作用的研究。

Targeting Inhibition of Notch1 Signaling Pathway on the Study of Human Gastric Cancer Stem Cells with Chemosensitization.

机构信息

Clinical Laboratory, Huaihe Hospital of Henan University, Kaifeng 475000, Henan, China.

出版信息

Comput Intell Neurosci. 2022 Apr 26;2022:1098394. doi: 10.1155/2022/1098394. eCollection 2022.

DOI:10.1155/2022/1098394

PMID:35515501

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9064537/

Abstract

BACKGROUND

Gastric cancer is the second most frequent cause of cancer death worldwide, although much geographical variation in incidence exists. Prevention and personalized treatment are regarded as the best options to reduce gastric cancer mortality rates (Hartgrink et al., 2009). Numerous studies have suggested that Notch1 and its ligands are overexpressed in gastric cancer, and its knockdown can inhibit the proliferation and survival of gastric cancer cells.

OBJECTIVE

To investigate the effect of Notch1 on the stemness and drug sensitivity of human gastric cancer SGC-7901 cells.

METHODS

Highly expressed Notch1 intracellular domain (NICD1) and Notch1-shRNA lentiviral expression vector were used to infect human gastric cancer SGC-7901 cells cultured in vitro, and western blot and immunofluorescence staining were used to identify highly expressed NICD and Notch1 silenced cells. The percentage of CD133+ cells was analyzed by flow cytometry, the expression of nestin and CFAP by immunofluorescence staining, the formation rate of tumor cell spheres and the tumorigenicity of SCID mice in vivo, and the regulation of cell stemness by Notch1. The sensitivity of each group of cells to the chemotherapeutic drugs teniposide (VM-26) and carmustine (BCNU) was also detected by the MTT method.

RESULTS

The stemness phenotype of tumor cells with the increased NICD expression was enhanced, such as an increased proportion of CD133+ cells, enhanced nestin expression, decreased GFAP expression, increased tumor cell sphere formation rate and tumorigenic rate of SCID mice implantation, and decreased sensitivity to VM-26 and BCNU. In contrast, the stemness phenotype of tumor cells with downregulated Notch1 gene expression was significantly suppressed, while the sensitivity to VM-26 and BCNU was increased.

CONCLUSION

High Notch1 expression increased the stemness of SGC-7901 cells and decreased the sensitivity of SGC-7901 cells to chemotherapeutic drugs.

摘要

背景

胃癌是全球第二大常见癌症死亡原因，尽管发病率存在很大的地域差异。预防和个性化治疗被认为是降低胃癌死亡率的最佳选择（Hartgrink 等人，2009 年）。许多研究表明，Notch1 及其配体在胃癌中过度表达，其敲低可以抑制胃癌细胞的增殖和存活。

目的

研究 Notch1 对人胃癌 SGC-7901 细胞干性和药物敏感性的影响。

方法

使用高表达 Notch1 胞内结构域（NICD1）和 Notch1-shRNA 慢病毒表达载体感染体外培养的人胃癌 SGC-7901 细胞，通过 Western blot 和免疫荧光染色鉴定高表达 NICD 和 Notch1 沉默细胞。通过流式细胞术分析 CD133+细胞的百分比，通过免疫荧光染色分析 nestin 和 CFAP 的表达，通过体内 SCID 小鼠肿瘤球形成率和肿瘤发生率，以及 Notch1 对细胞干性的调节。通过 MTT 法检测各组细胞对化疗药物替尼泊苷（VM-26）和卡莫司汀（BCNU）的敏感性。

结果

NICD 表达增加的肿瘤细胞干性表型增强，如 CD133+细胞比例增加、nestin 表达增强、GFAP 表达降低、肿瘤细胞球形成率和 SCID 小鼠植入瘤形成率增加、VM-26 和 BCNU 敏感性降低。相反，下调 Notch1 基因表达的肿瘤细胞干性表型显著受到抑制，而对 VM-26 和 BCNU 的敏感性增加。