Objective

To investigate the role of miR-374b in medicating biological function changes in cervical cancer cells by increasing the cytokine-induced killer (CIK) expression.

Methods

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.

Results

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.

Conclusion

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.

中文翻译：

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MiR-374b 通过靶向 PD-1/PD-L1 信号通路增加 CIK 表达并介导宫颈癌细胞生物学功能变化

客观的

研究 miR-374b 在通过增加细胞因子诱导的杀伤细胞 (CIK) 表达来治疗宫颈癌细胞生物学功能变化中的作用。

方法

对 62 名宫颈癌患者和 58 名健康人（包括人宫颈癌细胞系 (HeLa) 和正常人子宫平滑肌细胞 (HUSMC)）的静脉血进行了 miR-374b、PD-1 和 PD-L1 表达检测。将 sh-PD-1、si-PD-1、NC、miR-374b-inhibitor 和 miR-374b-mimics 转染到 HeLa 细胞中，通过以下方法测定 miR-374b、PD-1 和 PD-L1 的表达qRT-PCR检测,MTT检测和流式细胞仪分别检测细胞增殖和凋亡。

结果

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测定证实了 PD-1 和 miR-374b 之间的关系。转染 miR-374b-mimics 后，HeLa 细胞中 PD-1 的表达显着下调。与CIK+NC组相比，CIK+miR-374b-mimics组和CIK+siRNA-PD-1组PD-1的相对mRNA表达显着降低，与其他组相比活性显着降低HeLa细胞，两组均显示肿瘤体积明显缩小。

结论

MiR-374b通过靶向PD-1/PD-L1信号通路，增加CIK表达并介导宫颈癌细胞生物学功能变化，有望成为宫颈癌的潜在治疗靶点。