Zhang Yue, Xiao Yan-Wei, Ma Jing-Xin, Wang Ao-Xue
Department of Dermatology, the Second Hospital of Dalian Medical University, Dalian, Liaoning Province, 116021, China.
Department of Cell Biology, Dalian Medical University, Dalian, Liaoning Province, 116044, China.
Chin J Integr Med. 2024 Mar;30(3):213-221. doi: 10.1007/s11655-023-3607-2. Epub 2023 Sep 9.
To investigate the effect and possible mechanism of hydroxysafflor yellow A (HSYA) on human immortalized keratinocyte cell proliferation and migration.
HaCaT cells were treated with HSYA. Cell proliferation was detected by the cell counting kit-8 assay, and cell migration was measured using wound healing assay and Transwell migration assay. The mRNA and protein expression levels of heparin-binding epidermal growth factor (EGF)-like growth factor (HBEGF), EGF receptor (EGFR), phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), mammalian target of rapamycin (mTOR), and hypoxia-inducible factor-1α (HIF-1α) were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot, respectively. Circ_0084443-overexpressing HaCaT cells and empty plasmid HaCaT cells were constructed using the lentiviral stable transfection and treated with HSYA. The expression of circ_0084443 was detected by qRT-PCR.
HSYA (800 µmol/L) significantly promoted HaCaT cell proliferation and migration (P<0.05 or P<0.01). It also increased the mRNA and protein expression levels of HBEGF, EGFR, PI3K, AKT, mTOR and HIF-1α, and increased the phosphorylation levels of PI3K and AKT (P<0.05 or P<0.01). Furthermore, HSYA promoted HaCaT cell proliferation and migration via the HBEGF/EGFR and PI3K/AKT/mTOR signaling pathways (P<0.01). Circ_0084443 attenuated the mRNA expression levels of HBEGF, EGFR, PI3K, AKT, mTOR and HIF-1α (P<0.05). HSYA inhibited the circ_0084443 expression, further antagonized the inhibition of circ_0084443 on HBEGF, EGFR, PI3K, AKT, mTOR and HIF-1α, and promoted the proliferation of circ_0084443-overexpressing HaCaT cells (P<0.05 or P<0.01). However, HSYA could not influence the inhibitory effect of circ_0084443 on HaCaT cell migration (P>0.05).
HSYA played an accelerative role in HaCaT cell proliferation and migration, which may be attributable to activating HBEGF/EGFR and PI3K/AKT signaling pathways, and had a particular inhibitory effect on the keratinocyte negative regulator circ_0084443.
探讨羟基红花黄色素A（HSYA）对人永生化角质形成细胞增殖和迁移的影响及其可能机制。
用HSYA处理HaCaT细胞。采用细胞计数试剂盒-8法检测细胞增殖，用划痕愈合实验和Transwell迁移实验检测细胞迁移。分别通过定量实时聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法检测肝素结合表皮生长因子（HBEGF）、表皮生长因子受体（EGFR）、磷脂酰肌醇3激酶（PI3K）、蛋白激酶B（AKT）、雷帕霉素靶蛋白（mTOR）和缺氧诱导因子-1α（HIF-1α）的mRNA和蛋白表达水平。利用慢病毒稳定转染构建过表达circ_0084443的HaCaT细胞和空载质粒HaCaT细胞，并用HSYA处理。通过qRT-PCR检测circ_0084443的表达。
HSYA（800 μmol/L）显著促进HaCaT细胞增殖和迁移（P<0.05或P<0.01）。它还增加了HBEGF、EGFR、PI3K、AKT、mTOR和HIF-1α的mRNA和蛋白表达水平，并增加了PI3K和AKT的磷酸化水平（P<0.05或P<0.01）。此外，HSYA通过HBEGF/EGFR和PI3K/AKT/mTOR信号通路促进HaCaT细胞增殖和迁移（P<0.01）。Circ_0084443降低了HBEGF、EGFR、PI3K、AKT、mTOR和HIF-1α的mRNA表达水平（P<0.05）。HSYA抑制circ_0084443表达，进一步拮抗circ_0084443对HBEGF、EGFR、PI3K、AKT、mTOR和HIF-1α的抑制作用，并促进过表达circ_0084443的HaCaT细胞增殖（P<0.05或P<0.01）。然而，HSYA不影响circ_0084443对HaCaT细胞迁移的抑制作用（P>0.05）。
HSYA对HaCaT细胞增殖和迁移起促进作用，可能是通过激活HBEGF/EGFR和PI3K/AKT信号通路实现的，且对角质形成细胞负调控因子circ_0084443有特异性抑制作用。
