Du Ying, Ye Hui, Lin Mei, Cao Lili
Oncology Department, Shandong Provincial Qianfoshan Hospital, School of Medicine, Shandong University, Jinan, China.
Department of Pathology, School of Basic Medicine, Qingdao University, Qingdao, China.
Front Immunol. 2023 Dec 7;14:1267950. doi: 10.3389/fimmu.2023.1267950. eCollection 2023.
To explore the role of Krüppel-like factor 14 (KLF14) and its underlying mechanism(s) of action in cell-cycle regulation in cervical cancer.
Lentiviral infection was used to construct , zinc-finger structural mutations, and empty vector controls in SiHa and HeLa cervical cancer cells. The effect of KLF14 on cervical cancer cell cycle was detected by flow cytometry. The effect of KLF14 on the expression of cyclin-dependent kinase 2 (), cyclin A2 (CCNA2, and MAPK signalling pathway-related molecules was detected by fluorescence quantitative RT-PCR (qRT-PCR) and western blot. Cervical cancer cells were treated with JNK-pathway inhibitors/agonists before we assessed changes in the cell cycle and the expression of the CDK2, CCNA2, and p-JNK/JNK. Subcutaneous xenograft studies to explore the effects of KLF14 on cervical cancer cell proliferation , and western blotting was implemented to measure the expression of CCNA2, CDK2, and the activation levels of the MAPK-signaling pathway proteins in tumours.
The proportion of cells in the S phase was increased in the -overexpressing group compared with the control group (<0.001); CDK2, CCNA2, and -JNK/JNK expression levels were elevated in the overexpressing group relative to the control group (all <0.05). When JNK-pathway activation was inhibited/promoted, the proportion of cells in the S phase was reduced/increased (<0.05) and CDK2 and CCNA2 expression levels were reduced/decreased, respectively (all <0.05). Vivo experiments revealed that KLF14 inhibited cervical cancer cell proliferation (<0.01) and that -JNK/JNK, CDK2, and CCNA2 expression levels were augmented in tumours in the overexpression group (<0.01).
KLF14 induced S-phase arrest in cervical cancer cells and inhibited the proliferation of cervical cancer cells ; the induction of S-phase arrest was related to its zinc-finger structure. KLF14 also activated the JNK pathway to induce S-phase arrest and promote the expression of CDK2 and CCNA2. In summary, KLF14 activates the JNK-signaling pathway to induce S-phase arrest in cervical cancer cells.
探讨Krüppel样因子14（KLF14）在宫颈癌细胞周期调控中的作用及其潜在作用机制。
采用慢病毒感染法在SiHa和HeLa宫颈癌细胞中构建KLF14过表达、锌指结构突变体及空载体对照。通过流式细胞术检测KLF14对宫颈癌细胞周期的影响。采用荧光定量逆转录聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法检测KLF14对细胞周期蛋白依赖性激酶2（CDK2）、细胞周期蛋白A2（CCNA2）及丝裂原活化蛋白激酶（MAPK）信号通路相关分子表达的影响。在用JNK通路抑制剂/激动剂处理宫颈癌细胞后，评估细胞周期变化以及CDK2、CCNA2和磷酸化JNK/总JNK（p-JNK/JNK）的表达情况。通过皮下异种移植实验研究KLF14对宫颈癌细胞增殖的影响，并采用蛋白质免疫印迹法检测肿瘤组织中CCNA2、CDK2的表达以及MAPK信号通路蛋白的激活水平。
与对照组相比，KLF14过表达组S期细胞比例增加（P<0.001）；过表达组中CDK2、CCNA2及p-JNK/JNK的表达水平相对于对照组升高（均P<0.05）。当JNK通路激活被抑制/促进时，S期细胞比例降低/增加（P<0.05），CDK2和CCNA2的表达水平分别降低/降低（均P<0.05）。体内实验显示，KLF14抑制宫颈癌细胞增殖（P<0.01），过表达组肿瘤组织中p-JNK/JNK、CDK2和CCNA2的表达水平升高（P<0.01）。
KLF14诱导宫颈癌细胞发生S期阻滞并抑制宫颈癌细胞增殖；诱导S期阻滞与其锌指结构有关。KLF14还激活JNK通路以诱导S期阻滞并促进CDK2和CCNA2的表达。总之，KLF14激活JNK信号通路以诱导宫颈癌细胞发生S期阻滞。
