Jin Jian-Qiu, Wang Qian, Zhang Yu-Xing, Wang Xing, Lu Zhi-Yue, Li Bo-Wen
Department of Stomatology, Beijing Hospital, National Center of Gerontology, Beijing, People's Republic of China.
Institute of Geriatric Medicine, Chinese Academy of Medical Sciences, Beijing, 100730, People's Republic of China.
Lasers Med Sci. 2022 Dec;37(9):3461-3472. doi: 10.1007/s10103-022-03607-y. Epub 2022 Jul 7.
Early treatment of oral precancerous lesions is considered as a key strategy for in oral carcinogenesis prevention. Increasing evidence has suggested that the transforming growth factor beta (TGF-β) signaling pathway is tightly involved in the process of oral-carcinogenesis. In this study, we investigated the inhibition effect and potential mechanism of 5-aminolaevulinic acid photodynamic therapy (ALA-PDT) in human oral precancerous cells via TGF-β pathway.
Here, the dysplastic oral keratinocyte (DOK) cells were incubated with ALA concentration of 1 mM/mL for 4 h and then irradiated with a Helium-Neon (He-Ne) ion laser at 633 nm (200 mW/cm). The control cells were cultured in Dulbecco's modified Eagle's medium (DMEM) medium. We analyzed the differentially expressed genes and correlated pathways in oral precancerous cells following ALA-PDT using Affymetrix microarrays. TGF-β pathway was analyzed by quantitative real-time polymerase chain reaction (RT-qPCR) and western blotting. Bioinformatics analysis was performed to evaluate the expression of TGF-β1 in human oral cancer samples and adjacent normal samples. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), flow cytometry, 2'-7'-dichlorodihydrofluorescein diacetate (DCFH-DA), and wound healing assay were used to assess the effects of ALA-PDT plus TGF-β receptor inhibitor (LY2109761) in DOK cells.
The TGF-β signaling could exert in suppressive effects on DOK cells after ALA-PDT. The cell proliferation and migration rate of DOK cells was significantly reduced and apoptosis and ROS generation induced more effectively by ALA-PDT combined with LY2109761. Furthermore, cell cycle analysis revealed that the combined treatment resulted in G0/G1 phase arrest.
ALA-PDT suppresses the growth of oral precancerous cells by regulating the TGF-β signaling pathway, and its suppressive effect was enhanced using LY2109761. These results indicate that it could be a promising alternative treatment against oral precancerous lesions.
口腔癌前病变的早期治疗被认为是预防口腔癌发生的关键策略。越来越多的证据表明，转化生长因子β（TGF-β）信号通路紧密参与口腔癌发生过程。在本研究中，我们通过TGF-β通路研究了5-氨基乙酰丙酸光动力疗法（ALA-PDT）对人口腔癌前细胞的抑制作用及其潜在机制。
在此，将发育异常的口腔角质形成细胞（DOK）细胞与浓度为1 mM/mL的ALA孵育4小时，然后用波长633 nm（200 mW/cm）的氦氖（He-Ne）离子激光照射。对照细胞在杜氏改良 Eagle培养基（DMEM）中培养。我们使用Affymetrix微阵列分析了ALA-PDT后口腔癌前细胞中差异表达的基因及其相关通路。通过定量实时聚合酶链反应（RT-qPCR）和蛋白质印迹法分析TGF-β通路。进行生物信息学分析以评估TGF-β1在人口腔癌样本和相邻正常样本中的表达。使用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基-2H-四唑溴盐（MTT）、流式细胞术、2',7'-二氯二氢荧光素二乙酸酯（DCFH-DA）和伤口愈合试验评估ALA-PDT联合TGF-β受体抑制剂（LY2109761）对DOK细胞的影响。
ALA-PDT后，TGF-β信号可对DOK细胞发挥抑制作用。ALA-PDT联合LY2109761能更有效地降低DOK细胞的增殖和迁移率，并诱导细胞凋亡和活性氧生成。此外，细胞周期分析显示联合治疗导致G0/G1期阻滞。
ALA-PDT通过调节TGF-β信号通路抑制口腔癌前细胞的生长，并且使用LY2109761可增强其抑制作用。这些结果表明，它可能是一种有前景的治疗口腔癌前病变的替代方法。
