Tong Qiaoling, Hu Man, Deng Meixian
Department of Gynaecology, Wuhan Fifth Hospital, Wuhan, 430050, Hubei Province, China.
Wuhan Fifth Hospital.
Cell Mol Biol (Noisy-le-grand). 2022 Sep 30;68(9):63-67. doi: 10.14715/cmb/2022.68.9.10.
The purpose of this experiment was to observe the biological effect and mechanism of miR-10b on cervical cancer (CC) rats. For this purpose, the rat model of CC was established and divided into three groups (Inhibitors/ Mimics/Control). The miR-10b transfection efficiency was analyzed via RT-PCR in cervical tissues in each group. The content of CD3+, CD4+, and CD8+ was detected. The levels of IL-8, TNF-β, IL-6, (CAT, SOD, and MDA were determined via ELISA, and the apoptosis of cervical tissues was detected usingTUNEL assay. The expressions of Caspase-3, Bcl-2, and the mTOR/P70S6K pathway genes and proteins were detected by qRT-PCR and Western blotting. Results showed that miR-10b was significantly increased in the Mimics group and decreased in the Inhibitors group. The content of IL-8, TNF-β, IL-6, CAT and MDA was raised, while that of SOD notably declined in the Inhibitors group. There were remarkably more apoptotic cells in the Mimics group, dominated by gliocytes, and fewer apoptotic cells in the Inhibitors group, with increased content of CD3+, CD4+ and CD8+. The Bcl-2, mTOR, and P70S6K mRNA expressions in the Inhibitors group were up-regulated than those in the other two groups, and the Caspase-3 gene in the Mimics group was increased and close to that in the control group. In the Mimics group, the mTOR and P70S6K protein were remarkably lower than those in the Inhibitors group. In conclusion, miR-10b can inhibit the occurrence and development of CC in rats by suppressing mTOR/P70S6K signaling, reducing the level of inflammation and oxidative stress, and increasing the level of immune factors.
本实验旨在观察miR-10b对宫颈癌(CC)大鼠的生物学效应及其机制。为此,建立了CC大鼠模型并分为三组(抑制剂组/模拟物组/对照组)。通过RT-PCR分析每组宫颈组织中miR-10b的转染效率。检测CD3 +、CD4 +和CD8 +的含量。采用ELISA法测定IL-8、TNF-β、IL-6、(CAT、SOD和MDA的水平,采用TUNEL法检测宫颈组织的凋亡情况。通过qRT-PCR和蛋白质印迹法检测Caspase-3、Bcl-2以及mTOR/P70S6K信号通路基因和蛋白的表达。结果显示,模拟物组中miR-10b显著升高,抑制剂组中miR-10b显著降低。抑制剂组中IL-8、TNF-β、IL-6、CAT和MDA的含量升高,而SOD的含量显著下降。模拟物组中凋亡细胞明显增多,以神经胶质细胞为主,抑制剂组中凋亡细胞较少,CD3 +、CD4 +和CD8 +的含量增加。抑制剂组中Bcl-2、mTOR和P70S6K的mRNA表达上调,高于其他两组,模拟物组中Caspase-3基因增加且接近对照组。模拟物组中mTOR和P70S6K蛋白明显低于抑制剂组。综上所述,miR-10b可通过抑制mTOR/P70S6K信号通路、降低炎症和氧化应激水平以及提高免疫因子水平来抑制大鼠CC的发生和发展。