Background: Aloperine can regulate miR-296-5p/Signal Transducer and Activator of Transcription 3 (STAT3) pathway to inhibit the malignant development of colorectal cancer (CRC), but the regulatory mechanism is unclear. This study explored the upstream mechanism of Aloperine in reducing CRC damage from the perspective of the circRNA-miRNA-mRNA regulatory network.
Methods: After treatment with gradient concentrations of Aloperine (0.1 mmol/L, 0.2 mmol/L, 0.4 mmol/L, 0.8 mmol/L and 1 mmol/L) for 24 hours, changes in CRC cell proliferation and apoptosis were detected by functional experiments. Data of the differential expression of miR-296-5p in CRC patients and healthy people were obtained from Starbase. The effects of Aloperine on 12 differentially expressed circRNAs were detected. The binding of miR-296-5p with NOP2/Sun RNA methyltransferase 2 (circNSUN2) and STAT3 was predicted by TargetScan and confirmed through dual-luciferase experiments. The expressions of circNSUN2, miR-296-5p and STAT3 as well as apoptosis-related genes in CRC cells were detected by qRT-PCR and Western blot as needed. Rescue experiments were conducted to test the regulatory effects of circNSUN2, miR-296-5p and STAT3 on CRC cells.
Results: Aloperine at a concentration gradient inhibited proliferation and promoted apoptosis in CRC cells. The abnormally low expression of miR-296-5p in CRC could be upregulated by Aloperine. Among the differentially expressed circRNAs in CRC, only circNSUN2 not only targets miR-296-5p, but also can be regulated by Aloperine. The up-regulation of circNSUN2 offset the inhibitory effect of Aloperine on cancer cells. The rescue experiments finally confirmed the regulation of circNSUN2/miR-296-5p/STAT3 axis in CRC cells.
Conclusion: By regulating the circNSUN2/miR-296-5p/STAT3 pathway, Aloperine prevents the malignant development of CRC cells.



中文翻译：

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Aloperine 通过调节 circNSUN2/miR-296-5p/STAT3 通路抑制结直肠癌细胞增殖和促进凋亡

背景： Aloperine 可通过调节 miR-296-5p/Signal Transducer and Activator of Transcription 3 (STAT3) 通路抑制结直肠癌 (CRC) 的恶性发展，但其调节机制尚不清楚。本研究从circRNA-miRNA-mRNA调控网络的角度探讨了Aloperine减少CRC损伤的上游机制。
方法：用梯度浓度的Aloperine（0.1 mmol/L、0.2 mmol/L、0.4 mmol/L、0.8 mmol/L和1 mmol/L）处理24 h后，通过功能实验检测CRC细胞增殖和凋亡的变化。miR-296-5p在CRC患者和健康人中的差异表达数据来自Starbase。检测了 Aloperine 对 12 个差异表达的 circRNA 的影响。TargetScan 预测 miR-296-5p 与 NOP2/Sun RNA 甲基转移酶 2 (circNSUN2) 和 STAT3 的结合，并通过双荧光素酶实验证实。根据需要采用qRT-PCR和Western blot检测CRC细胞中circNSUN2、miR-296-5p和STAT3以及凋亡相关基因的表达。进行抢救实验以测试circNSUN2、miR-296-5p和STAT3对CRC细胞的调节作用。
结果：浓度梯度的 Aloperine 在 CRC 细胞中抑制增殖并促进细胞凋亡。Aloperine 可上调 CRC 中 miR-296-5p 的异常低表达。在CRC中差异表达的circRNA中，只有circNSUN2不仅靶向miR-296-5p，而且可以被Aloperine调控。circNSUN2的上调抵消了Aloperine对癌细胞的抑制作用。救援实验最终证实了 CRC 细胞中 circNSUN2/miR-296-5p/STAT3 轴的调控。
结论： Aloperine通过调控circNSUN2/miR-296-5p/STAT3通路，阻止CRC细胞的恶性发展。