Han Yujiao, Lian Ling, Ren Man, Li Shenghe, Zhao Chunfang, Jin Erhui
College of Animal Science, Anhui Science and Technology University, Chuzhou, China.
Anhui Province Key Laboratory of Animal Nutritional Regulation and Health, Chuzhou, China.
Ann Transl Med. 2022 Aug;10(16):873. doi: 10.21037/atm-22-3519.
Marek's disease (MD), a class II infectious, lymphoproliferative disease that mainly afflicts poultry, has been shown to cause wasting, limb paralysis, and often acute death. It is a neoplastic disease caused by a cell-binding herpesvirus that leads to the formation of tumors in various organs and tissues. Our previous reports have found that the microRNA, gga-miR-29b-3p, showed abnormal expression in MD lymphoma. However, it remains unknown whether gga-miR-29b-3p affects MD tumorigenesis.
The MD tumor cell line MSB1 was chosen to analyze the characteristics of gga-miR-29b-3p in tumors. Cell proliferation and migration were assessed by Cell Counting Kit-8 (CCK-8) and Transwell, respectively, and cell apoptosis and cycle were analyzed via fluorescent staining and flow cytometry, respectively. The regulation between gga-miR-29b-3p and its potential target genes was verified by dual luciferase results and loss-of-function assays. The effect of target genes was verified by examining the degree of RNA interference on MSB1 cells.
Analysis revealed that gga-miR-29b-3p impaired the proliferation of the MSB1 MD tumor cell line, induced apoptosis without obvious effects on the cell cycle, and suppressed the expression of the invasion-associated and genes. It was concluded that is the direct target of gga-miR-29b-3p. As expected, the effects of knockdown with small interfering RNA (siRNA) on MSB1 cell proliferation, apoptosis, and cycle were associated with gga-miR-29b-3p overexpression. Moreover, and were downregulated and was upregulated in both the gga-miR-29b-3p overexpression and knockdown groups. The expression levels of invasion-related genes were decreased post- knockdown. In both the gga-miR-29b-3p overexpression and knockdown conditions, a decrease in oncogene expression in MD virus was observed.
Overall, gga-miR-29b-3p was demonstrated to have a suppressive effect in MD lymphoma progression via the targeting of the gene. Gga-miR-29b-3p overexpression and knockdown inhibited MSB1 cell proliferation through suppressing the pro-apoptotic gene expression and elevating the anti-apoptotic gene expression in the apoptosis pathway. Our study provides a theoretical basis for targeted treatment of MD.
马立克氏病（MD）是一种主要感染家禽的II类传染性淋巴增生性疾病，已被证明会导致消瘦、肢体麻痹，并常常引发急性死亡。它是一种由细胞结合性疱疹病毒引起的肿瘤性疾病，可导致各个器官和组织中形成肿瘤。我们之前的报告发现，微小RNA gga-miR-29b-3p在MD淋巴瘤中表达异常。然而，gga-miR-29b-3p是否影响MD肿瘤发生仍不清楚。
选择MD肿瘤细胞系MSB1来分析gga-miR-29b-3p在肿瘤中的特性。分别通过细胞计数试剂盒-8（CCK-8）和Transwell评估细胞增殖和迁移，通过荧光染色和流式细胞术分别分析细胞凋亡和细胞周期。通过双荧光素酶结果和功能丧失实验验证gga-miR-29b-3p与其潜在靶基因之间的调控关系。通过检测RNA干扰对MSB1细胞的影响程度来验证靶基因的作用。
分析表明，gga-miR-29b-3p损害了MSB1 MD肿瘤细胞系的增殖，诱导细胞凋亡，对细胞周期无明显影响，并抑制侵袭相关基因 和 的表达。得出结论， 是gga-miR-29b-3p的直接靶标。正如预期的那样，用小干扰RNA（siRNA）敲低 对MSB1细胞增殖、凋亡和细胞周期的影响与gga-miR-29b-3p过表达相关。此外，在gga-miR-29b-3p过表达组和 敲低组中， 和 下调， 上调。敲低 后侵袭相关基因的表达水平降低。在gga-miR-29b-3p过表达和 敲低条件下，均观察到MD病毒中 癌基因表达下降。
总体而言，通过靶向 基因，gga-miR-29b-3p在MD淋巴瘤进展中具有抑制作用。gga-miR-29b-3p过表达和 敲低通过抑制凋亡途径中促凋亡基因表达和提高抗凋亡基因表达来抑制MSB1细胞增殖。我们的研究为MD的靶向治疗提供了理论依据。
