Department of Gynecologic Oncology, Jiangxi Cancer Hospital, Nanchang, 330006, China.
Department of Gynaecology, No. 1 Hospital of Zibo, Zibo, 255200, China.
Cell Mol Biol (Noisy-le-grand). 2022 Feb 4;67(5):131-137. doi: 10.14715/cmb/2021.67.5.18.
This study focused on miR-494 inhibiting the proliferation, migration and invasion of cervical cancer cells by regulating HCCR1. During the study, 30 pairs of primary cervical cancer tissues and adjacent tissues diagnosed by pathology were collected, and then placed in a cryopreservation tube, immediately placed in liquid nitrogen to freeze, and then moved to a -80 °C refrigerator for storage until use. They were dipped with crystal violet staining solution for 20 minutes, washed 3 times with PBS, and dried at room temperature. The polycarbonate film on the chamber was gently cut, placed on a glass slide, and covered with neutral resin. The number of cells in 5 random fields was counted under a microscope, and the differences between the groups were compared. The qRT-PCR results showed that compared with the normal cervical tissue cell line HCCR1, the expression levels of miR-494 in cervical cancer cell lines HCCR1 and C-33A were significantly reduced. Compared with the transfection of mimics NC, the level of HCCR1mRNA and protein decreased significantly when transfected with miRNA-494mimics (P <0.05); compared with the transfection of inhibitor NC, the level of HCCR1 mRNA and protein increased significantly when transfected with miRNA-494 inhibitor (P <0.05).
本研究通过调控 HCCR1 来聚焦 miR-494 抑制宫颈癌细胞的增殖、迁移和侵袭。研究过程中,收集了 30 对经病理诊断的原发性宫颈癌组织及癌旁组织,然后将其放入冻存管中,立即置于液氮中冷冻,再转移至-80°C冰箱中保存备用。将其用结晶紫染色液浸染 20 分钟,再用 PBS 洗 3 次,室温下干燥。轻轻切下小室上的聚碳酸酯膜,将其放在载玻片上,并用中性树脂覆盖。在显微镜下对 5 个随机视野中的细胞计数,并比较组间差异。qRT-PCR 结果显示,与正常宫颈组织细胞系 HCCR1 相比,宫颈癌细胞系 HCCR1 和 C-33A 中 miR-494 的表达水平显著降低。与转染 mimics NC 相比,转染 miRNA-494mimics 后 HCCR1mRNA 和蛋白水平显著降低(P<0.05);与转染 inhibitor NC 相比,转染 miRNA-494 抑制剂后 HCCR1mRNA 和蛋白水平显著升高(P<0.05)。