Yang Haiyan, Fang Ziyu, Qu Xiaoli, Zhang Xiaoli, Wang Yifeng
Department of Obstetrics and Gynecology, Fourth Affiliated Hospital of Guangxi Medical University, Liuzhou, Guangxi, 545005, People's Republic of China.
Cancer Manag Res. 2020 Jan 22;12:497-509. doi: 10.2147/CMAR.S226547. eCollection 2020.
Evidence suggested that procyanidin compound (PC) could inhibit the progression of cervical cancer (CC); however, the mechanism still remains unclear. We aimed to study the potential mechanism of PC acting on CC cells.
After a 24 hr incubation of lipopolysaccharide (LPS) (1 μg/mL), human CC SiHa and HeLa cells were cultured with various concentrations (20, 40, and 80 μg/mL) of PC for 24 hrs, then the cell viability was detected using Cell Counting Kit-8 (CCK-8). The migration and invasion abilities were assessed by scratch and Transwell assays. Apoptosis and cell cycle were detected using flow cytometry. Real-time quantitative PCR (RT-qPCR) and Western blot were used for expression analysis of the inflammatory cytokines. The pathway components were measured to evaluate the involvement of toll-like receptor 4/nuclear factor kappa-light-chain-enhancer of activated B cells (TLR4/NF-κB) pathway.
PC inhibited the LPS-primed cell viability in a dose-dependent manner. After PC treatment, cell migration and invasion were inhibited, cell number at the G2/M phase was increased. The CC cell apoptosis was triggered through upregulating levels of cleaved caspase-3 and Bax and downregulating the level of B-cell lymphoma 2 protein. A significant reduction was shown in the levels of interleukin (IL)-6, IL-1β and tumor necrosis factor (TNF)-α. Furthermore, a remarkable reduction in the ratio of TLR4 and the p-P65/t-P65 and in the progression of P65 translocation into the nucleus was observed.
Our results revealed that the inhibitory effect of PC on CC cell proliferation relies on the induction of apoptosis and inhibition of inflammatory cytokines.
有证据表明原花青素化合物（PC）可抑制宫颈癌（CC）的进展；然而，其机制仍不清楚。我们旨在研究PC作用于CC细胞的潜在机制。
用脂多糖（LPS）（1μg/mL）孵育24小时后，将人CC SiHa和HeLa细胞与不同浓度（20、40和80μg/mL）的PC培养24小时，然后使用细胞计数试剂盒-8（CCK-8）检测细胞活力。通过划痕试验和Transwell试验评估迁移和侵袭能力。使用流式细胞术检测细胞凋亡和细胞周期。采用实时定量PCR（RT-qPCR）和蛋白质免疫印迹法进行炎症细胞因子的表达分析。检测信号通路成分以评估Toll样受体4/活化B细胞核因子κB（TLR4/NF-κB）信号通路的参与情况。
PC以剂量依赖的方式抑制LPS引发的细胞活力。PC处理后，细胞迁移和侵袭受到抑制，G2/M期细胞数量增加。通过上调裂解的半胱天冬酶-3和Bax的水平并下调B细胞淋巴瘤2蛋白的水平，触发CC细胞凋亡。白细胞介素（IL）-6、IL-1β和肿瘤坏死因子（TNF）-α水平显著降低。此外，观察到TLR4与p-P65/t-P65的比例以及P65向细胞核转位的进程显著降低。
我们的结果表明，PC对CC细胞增殖的抑制作用依赖于诱导细胞凋亡和抑制炎症细胞因子。
