Department of Gynecology, The First People's Hospital of Lianyungang (Xuzhou Medical University Affiliated Hospital of Lianyungang), (The First Affiliated Hospital of Kangda College of Nanjing Medical University), Lianyungang, 222000, China.
Department of Gynecology, The First People's Hospital of Lianyungang (Xuzhou Medical University Affiliated Hospital of Lianyungang), (The First Affiliated Hospital of Kangda College of Nanjing Medical University), Lianyungang, 222000, China.
J Reprod Immunol. 2021 Feb;143:103265. doi: 10.1016/j.jri.2020.103265. Epub 2020 Dec 8.
To investigate the role of miR-374b in medicating biological function changes in cervical cancer cells by increasing the cytokine-induced killer (CIK) expression.
Venous blood of 62 cervical cancer patients and 58 healthy individuals including Human cervical cancer cell line (HeLa) and normal human uterine smooth muscle cells (HUSMC) were tested for expression of miR-374b, PD-1, and PD-L1. sh-PD-1, si-PD-1, NC, miR-374b-inhibitor, and miR-374b-mimics were transfected into HeLa cells, and expression of miR-374b, PD-1, and PD-L1 was determined by a qRT-PCR assay, and the proliferation and apoptosis of the cells were detected using a MTT assay and flow cytometry, respectively.
PD-1 was highly expressed in cervical cancer, while miR-374b is lowly expressed in it, and the area-under-the-curve (AUC) of both PD-1 and miR-374b was larger than 0.8. The dual luciferase reporter assay confirmed relationship between PD-1 and miR-374b. Expression of PD-1 in HeLa cells was significantly down-regulated after transfection of miR-374b-mimics. Compared with the CIK + NC group, the CIK + miR-374b-mimics group and the CIK + siRNA-PD-1 group showed a significant decrease in the relative mRNA expression of PD-1, compared with other group showed significantly lowered activity of HeLa cells, and the two groups showed significantly reduced tumor volume.
MiR-374b increases the CIK expression and mediates biological function changes in cervical cancer cells by targeting the PD-1/PD-L1 signaling pathway, so it is expected to be a potential therapeutic target for cervical cancer.
通过增加细胞因子诱导的杀伤细胞（CIK）的表达来研究 miR-374b 在介导宫颈癌细胞生物学功能变化中的作用。
检测 62 例宫颈癌患者和 58 例健康个体（包括人宫颈癌细胞系（HeLa）和正常人类子宫平滑肌细胞（HUSMC））的 miR-374b、PD-1 和 PD-L1 的表达。将 sh-PD-1、si-PD-1、NC、miR-374b 抑制剂和 miR-374b 模拟物转染到 HeLa 细胞中，通过 qRT-PCR 检测 miR-374b、PD-1 和 PD-L1 的表达，分别通过 MTT 检测和流式细胞术检测细胞的增殖和凋亡。
PD-1 在宫颈癌中高表达，而 miR-374b 在宫颈癌中低表达，PD-1 和 miR-374b 的曲线下面积（AUC）均大于 0.8。双荧光素酶报告基因实验证实了 PD-1 和 miR-374b 之间的关系。转染 miR-374b 模拟物后 HeLa 细胞中 PD-1 的表达明显下调。与 CIK+NC 组相比，CIK+miR-374b 模拟物组和 CIK+siRNA-PD-1 组的 PD-1 相对 mRNA 表达明显降低，与其他组相比，HeLa 细胞的活性明显降低，两组的肿瘤体积明显减小。
miR-374b 通过靶向 PD-1/PD-L1 信号通路增加 CIK 的表达并介导宫颈癌细胞的生物学功能变化，因此有望成为宫颈癌的潜在治疗靶点。
