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脂肪间充质干细胞分泌组联合呼肠孤病毒与单纯联合呼肠孤病毒对 CT26 细胞的作用比较。

Comparison of the Effect of Adipose Mesenchymal Stem Cells-Derived Secretome with and without Reovirus in CT26 Cells.

机构信息

Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.

Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.

出版信息

Arch Razi Inst. 2022 Apr 30;77(2):615-622. doi: 10.22092/ARI.2021.353845.1615. eCollection 2022 Apr.

Abstract

Colorectal cancer is the fourth leading cause of cancer-related deaths that has significantly increased over the past three decades. New therapeutic approaches, such as oncolytic viruses, have become very imperative recently to destroy cancer cells. The use of mesenchymal stem cells (MSCs) secretome that is produced in response to variant conditions involves different paracrine molecules secretion that has therapeutic potential in several chronic diseases. Mesenchymal stem cells and their derivatives are employed as regenerative medicine; nevertheless, there is ambiguity in the function of these cells in the control of malignancy. This study aimed to examine the apoptotic effect of secretomes derived from MSCs affected by encompassing oncolytic reoviruses. Mesenchymal stem cells were cultured after separation from abdominal adipose tissue of BALB/c mice. After three passages, the cells were infected by reovirus at the multiplicity of infection of 1 plaque-forming unit per cell. Uninfected and infected secretomes with reovirus were collected separately. The colorectal cancer CT26 cells were confronted with uninfected secretome, infected secretions, reovirus as a positive control, and Dulbecco's Modified Eagle Medium/High Glucose as negative control separately. Finally, apoptosis and necrosis were evaluated by flow cytometry. The infected secretome with reovirus was capable to induce apoptosis more than the uninfected secretome in CT26. However, the supernatant of reovirus infected cells was more capable to induce cell death, in comparison to the infected secretome. Infected MSCs with oncolytic reovirus produced a type of condition media that enhanced apoptosis induction and could have a therapeutic effect on cancer cells. Nonetheless, tumoral cells confronted with the oncolytic reovirus showed more capability in inducing apoptosis in CT26 cells. As a result, the use of oncolytic virus and infected secretome are more effective than uninfected secretome in inducing apoptosis.

摘要

结直肠癌是癌症相关死亡的第四大主要原因,在过去三十年中显著增加。最近,新的治疗方法,如溶瘤病毒,已成为破坏癌细胞的非常必要手段。间质干细胞(MSCs)分泌的外泌体是对不同条件的反应,涉及不同的旁分泌分子分泌,在几种慢性疾病中有治疗潜力。间质干细胞及其衍生物被用作再生医学;然而,这些细胞在控制恶性肿瘤方面的功能尚不清楚。本研究旨在研究受包裹溶瘤呼肠孤病毒影响的 MSC 分泌组的凋亡作用。从 BALB/c 小鼠腹部脂肪组织中分离出间质干细胞后进行培养。经过三代传代,细胞以每细胞 1 个蚀斑形成单位的感染复数被呼肠孤病毒感染。分别收集未感染和感染呼肠孤病毒的分泌组。将结肠直肠癌细胞 CT26 分别与未感染的分泌组、感染的分泌组、呼肠孤病毒(阳性对照)和 Dulbecco 改良 Eagle 培养基/高葡萄糖(阴性对照)接触。最后,通过流式细胞术评估细胞凋亡和坏死。与未感染的分泌组相比,感染呼肠孤病毒的分泌组能够更有效地诱导 CT26 细胞凋亡。然而,与感染的分泌组相比,呼肠孤病毒感染细胞的上清液更能诱导细胞死亡。携带溶瘤呼肠孤病毒的感染 MSC 产生了一种增强细胞凋亡诱导的条件培养基,可能对癌细胞具有治疗作用。尽管如此,与溶瘤呼肠孤病毒接触的肿瘤细胞在 CT26 细胞中显示出更强的诱导凋亡能力。因此,与未感染的分泌组相比,溶瘤病毒和感染的分泌组在诱导细胞凋亡方面更有效。

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