Department of Gynecology, Nantong Tumor Hospital,Nantong 226001,Jiangsu,China.
Cell Mol Biol (Noisy-le-grand). 2020 Oct 31;66(7):180-185.
In recent years, studies have found that miR-RNA plays a role in cell proliferation, differentiation, apoptosis and metabolism. Among them, miR-196a is closely related to cervical cancer. Therefore, this experiment investigates the effect of mir-196a expression on cervical cancer cells and related mechanisms. The expression level of miR-196a in the cervical cancer cell line was assayed with the RT-PCR method, and liposome transfection was used to investigate its up-regulation or down-regulation in cervical cancer cells. The CCK-8 method and flow cytometry were used to measure cervical cancer cell proliferation and apoptosis, while the Transwell assay was used to determine cell migration and invasion of each transfection group. Bioinformatics was used to predict the target gene of miR-196a, which was verified using dual luciferase report experiment and Western blot, and miR-196a was further transfected with si-LRIG3 to detect its reversal effect on miR-196a regulation. Inhibition of the expression of miR-196a significantly reduced the proliferation, migration and invasion of cervical cancer cells, and promoted their apoptosis. Results from dual luciferase assay showed that miR-196a and LRIG3 had direct targeting effects. Cell proliferation, migration and invasion were enhanced by a reduction in the expression level of LRIG3 protein after miR-196a inhibitor cells were transfected with si-LRIG3. The expression of miR-196a is up-regulated in cervical cancer, and it promotes the growth of cervical cancer by its targeting effect on LRIG3 expression, resulting in enhancement of the proliferation, migration and invasion ability of cervical cancer cells, and inhibition of apoptosis.
近年来的研究发现,miR-RNA 在细胞增殖、分化、凋亡和代谢中发挥作用。其中,miR-196a 与宫颈癌密切相关。因此,本实验研究了 mir-196a 表达对宫颈癌细胞的影响及相关机制。采用 RT-PCR 方法检测宫颈癌细胞系中 miR-196a 的表达水平,并用脂质体转染法研究其在宫颈癌细胞中的上调或下调。CCK-8 法和流式细胞术检测宫颈癌细胞增殖和凋亡,Transwell 小室法检测各组转染细胞的迁移和侵袭。生物信息学预测 miR-196a 的靶基因,并用双荧光素酶报告实验和 Western blot 进行验证,进一步用 si-LRIG3 转染 miR-196a 检测其对 miR-196a 调控的逆转作用。抑制 miR-196a 的表达显著降低了宫颈癌细胞的增殖、迁移和侵袭能力,并促进了其凋亡。双荧光素酶实验结果表明,miR-196a 与 LRIG3 具有直接的靶向作用。miR-196a 抑制剂转染 si-LRIG3 后,LRIG3 蛋白表达水平降低,细胞增殖、迁移和侵袭能力增强。miR-196a 在宫颈癌中呈上调表达,通过对 LRIG3 表达的靶向作用促进宫颈癌的生长,从而增强宫颈癌细胞的增殖、迁移和侵袭能力,抑制凋亡。