Hsa_circRNA_001859 通过 miR-21-5p/SLC38A2 通路调控胰腺癌进展和上皮间质转化。
Hsa_circRNA_001859 regulates pancreatic cancer progression and epithelial-mesenchymal transition through the miR-21-5p/SLC38A2 pathway.
机构信息
Department of General Surgery, The Second Affiliated Hospital of Xinjiang Medical University, Urumqi, Xinjiang, China.
Department of Pancreatic Surgery, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, Xinjiang, China.
出版信息
Cancer Biomark. 2023;37(1):39-52. doi: 10.3233/CBM-220229.
OBJECTIVE
This study attempts to investigate whether hsa_circRNA_001859 (circ_001859) could regulate the proliferation and invasion of pancreatic cancer through the miR-21-5p/SLC38A2 pathway.
METHODS
GSE79634 microarray was analyzed with R package. The expression of circ_001859 in pancreatic cancer tissues and cells was verified by qRT-PCR. After the overexpression of circ_001859, cell proliferation, cell migration and invasion were verified by colony formation and transwell assay. The targeting relationship between miR-21-5p and circ_001859 was predicted by TargetScan and was verified by dual luciferase reporter assay, RNA pull down and qRT-PCR. The effect of miR-21-5p on cell proliferation, migration and invasion were investigated by colony formation and transwell assay respectively. Similarly, the targeting relationship between miR-21-5p and SLC38A2 was predicted by TargetScan and was verified by dual luciferase reporter assay, western blot and qRT-PCR. The effect of SLC38A2 on cell proliferation was investigated by colony formation.
RESULTS
Circ_001859 was lowly expressed in pancreatic cancer tissues and cells. In vitro assays showed that overexpression of circ_001859 could inhibit the proliferation, migration and invasion of pancreatic cancer. In addition, this effect was also confirmed in xenograft transplantation model. Circ_001859 could be bind to miR-21-5p and sponge its expression in pancreatic cancer cells. Overexpression of miR-21-5p enhanced the proliferation, migration and invasion ability of pancreatic cancer cells, while the inhibition of miR-21-5p expression suppressed these abilities. Moreover, miR-21-5p directly targeted at SLC38A2 and inhibited SLC38A2 expression levels while circ_001859 up-regulated SLC38A2 levels. SLC38A2 expression knockdown enhanced cell proliferation but SLC38A2 overexpression resulted in decreased proliferation, and effects of SLC38A2 could be rescued by miR-21-5p and circ_001859. In addition, both QRT-PCR and immunofluorescence confirmed that circ_001859 could regulate tumor epithelial-mesenchymal transition (EMT) through the miR-21-5p/SLC38A2 pathway.
CONCLUSIONS
This study suggests that circ_001859 may inhibit the proliferation, invasion and EMT of pancreatic cancer through the miR-21-5p/SLC38A2 pathway.
目的
本研究旨在探讨 hsa_circRNA_001859（circ_001859）是否可以通过 miR-21-5p/SLC38A2 通路调节胰腺癌的增殖和侵袭。
方法
使用 R 软件包对 GSE79634 微阵列进行分析。通过 qRT-PCR 验证胰腺癌组织和细胞中 circ_001859 的表达。过表达 circ_001859 后，通过集落形成和 Transwell 分析验证细胞增殖、细胞迁移和侵袭。通过 TargetScan 预测 miR-21-5p 与 circ_001859 的靶向关系，并通过双荧光素酶报告基因分析、RNA 下拉和 qRT-PCR 进行验证。通过集落形成和 Transwell 分析分别研究 miR-21-5p 对细胞增殖、迁移和侵袭的影响。同样，通过 TargetScan 预测 miR-21-5p 与 SLC38A2 的靶向关系，并通过双荧光素酶报告基因分析、western blot 和 qRT-PCR 进行验证。通过集落形成研究 SLC38A2 对细胞增殖的影响。
结果
circ_001859 在胰腺癌组织和细胞中低表达。体外实验表明，过表达 circ_001859 可抑制胰腺癌的增殖、迁移和侵袭。此外，在异种移植移植模型中也得到了证实。Circ_001859 可以与 miR-21-5p 结合并海绵其在胰腺癌细胞中的表达。过表达 miR-21-5p 增强了胰腺癌细胞的增殖、迁移和侵袭能力，而抑制 miR-21-5p 表达则抑制了这些能力。此外，miR-21-5p 直接靶向 SLC38A2 并抑制 SLC38A2 表达水平，而 circ_001859 上调 SLC38A2 水平。SLC38A2 表达下调增强了细胞增殖，但 SLC38A2 过表达导致增殖减少，并且 SLC38A2 的表达可以通过 miR-21-5p 和 circ_001859 得到挽救。此外，qRT-PCR 和免疫荧光证实，circ_001859 可以通过 miR-21-5p/SLC38A2 通路调节肿瘤上皮-间质转化（EMT）。
结论
本研究表明，circ_001859 可能通过 miR-21-5p/SLC38A2 通路抑制胰腺癌的增殖、侵袭和 EMT。
Zotero 插件