背景:基于铂的化合物通常用作结直肠癌(CRC)的初始治疗。然而,CRC患者耐药性的发展需要在临床治疗期间给予高浓度的药物,从而增加铂基化合物的毒性并增加死亡率。STAG2是许多癌症中一个显著相关的耐药基因,但尚未在结直肠癌中进行研究。因此,本研究旨在探讨顺铂耐药基因STAG2的作用及药物敏感性。
方法:使用癌症药物敏感性基因组学(GDSC)和Kaplan-Meier(KM)绘图仪数据库检查了STAG2对CRC患者耐药性和生存率的影响。随后,sh-STAG2-HT-29细胞系使用STAG2的敲低测试产生,并且两种细胞系的半最大抑制浓度(IC50)使用细胞活力测试确定。然后我们使用了各种技术,包括细胞计数套件-8(CCK-8),平板克隆,5-乙炔基-2'-脱氧尿苷(EdU)荧光染色,流式细胞术检测细胞周期,疤痕试验,Transwell入侵试验,膜联蛋白V-异硫氰酸荧光素(FITC)/碘化丙啶(PI)荧光染色检测细胞凋亡,研究癌细胞系四个亚组的功能。此外,蛋白质印迹(WB)用于鉴定与观察到的功能改变相关的潜在途径。最后,表型,肿瘤重量,小鼠体重,肿瘤体积,使用皮下肿瘤形成方法评估发展的肿瘤的肿瘤组织结构。
结果:数据库分析表明,STAG2在促进CRC患者的耐药性中起作用。此外,该基因的突变导致对顺铂的敏感性增加,其过度表达与不良预后相关。随着STAG2敲低细胞的成功发展,在HT-29和sh-STAG2-HT-29细胞之间观察到IC50浓度的差异。选择10μM顺铂的治疗浓度,和扩散,迁移,STAG2敲低后癌细胞的侵袭能力下降。此外,细胞对顺铂治疗的敏感性增加,这可能是由上皮间质转化(EMT)途径介导的。在老鼠身上,通过STAG2敲低,HT-29细胞的致瘤潜能降低,伴随着对顺铂治疗的耐药性降低。
结论:STAG2在CRC中充当原癌基因,且其对顺铂治疗的耐药性更为突出。本研究证实了STAG2在CRC中的作用,并为进一步开发STAG2作为患者接受铂类药物治疗时确定剂量的辅助标准提供了理论依据。
BACKGROUND: Platinum-based compounds are commonly used as an initial treatment for colorectal cancer (CRC). However, the development of drug resistance in patients with CRC necessitates the administration of high drug concentrations during clinical treatment, thereby augmenting the toxicity of platinum-based compounds and increasing the mortality rate.
STAG2 is a significantly associated drug-resistance gene in many cancers, but it has not been studied in colorectal cancer. Therefore, the present study aimed to investigate the role and drug sensitivity of the cisplatin-resistant gene
STAG2.
METHODS: The effects of
STAG2 on drug resistance and survival rates of patients with CRC were examined using the Genomics of Drug Sensitivity in Cancer (GDSC) and Kaplan-Meier (KM) plotter databases. Subsequently, a sh-
STAG2-HT-29 cell line was generated using a knockdown test of
STAG2, and the half-maximal inhibitory concentration (IC50) of the two cell lines was determined using a cell viability test. We then used various techniques, including the Cell Counting Kit-8 (CCK-8), plate cloning, 5-ethynyl-2\'-deoxyuridine (EdU) fluorescence staining, flow cytometry for cell cycle detection, the scar assay, the Transwell invasion assay, and Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) fluorescence staining for apoptosis detection, to investigate the functionality of the four subgroups of cancer cell lines. Additionally, Western blotting (WB) was used to identify the potential pathways associated with the observed functional alterations. Finally, the phenotype, tumor weight, mouse weight, tumor volume, and tumor tissue structure of the developed tumors were assessed using the subcutaneous tumor formation method.
RESULTS: Database analysis indicated that STAG2 plays a role in facilitating drug resistance among individuals with CRC. Furthermore, mutations in this gene lead to increased sensitivity to cisplatin, and its overexpression was associated with an unfavorable prognosis. Following the successful development of STAG2 knockdown cells, differences in IC50 concentrations were observed between HT-29 and sh-
STAG2-HT-29 cells. A treatment concentration of 10 μM cisplatin was selected, and the proliferation, migration, and invasion capabilities of cancer cells decreased after STAG2 knockdown. Additionally, the sensitivity of the cells to cisplatin therapy was increased, which was potentially mediated by the epithelial-mesenchymal transition (EMT) pathway. In mice, the tumorigenic potential of HT-29 cells was reduced by STAG2 knockdown, accompanied by a decrease in resistance to cisplatin therapy.
CONCLUSIONS: STAG2 acts as a proto-oncogene in CRC, and its resistance to cisplatin therapy is more prominent. This study confirmed the role of STAG2 in CRC and provided a theoretical basis for the further development of STAG2 as an auxiliary criterion for determining dosage when patients are treated with platinum drugs.