Chen X, Bai J, Li Y, Li X-G, Lv G-F, Zhang H-Ch
School of Pharmacy, , , China.
, , China.
Genet Mol Res. 2017 May 31;16(2):gmr-16-02-gmr.16029430. doi: 10.4238/gmr16029430.
Hypoxia-inducible factor-1α (HIF-1α) is considered the main transcriptional regulator of the hypoxia-specific cellular and developmental response. This study was performed to investigate the effect of Shenqin biochemical extract (SQBE) on HIF-1α expression in ultraviolet B (UVB)-irradiated HaCaT cells and the possible action mechanisms of SQBE against UVB-induced skin cancer. HaCaT cells in logarithmic growth phase were seeded in Dulbecco's modified Eagle's medium with 10% fetal bovine serum, and conventionally cultured at 37°C with 5% CO. Cells were divided into control group (administered the same amounts of dimethyl sulfoxide), SQBE1 group (12.5 μg/mL SQBE), SQBE2 group (25.0 μg/mL SQBE), and SQBE3 group (50.0 μg/mL SQBE). Four hours post administration, the control and treatment groups were irradiated with UVB (0, 20, 40, and 60 mJ/cm). After 24 h, cell survival rate was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Expression levels of HIF-1α mRNA and protein were detected by polymerase chain reaction and western blotting, respectively. SQBE-treated, UVB-irradiated cells had improved survival rates. This increase was most significant in SQBE3 group (P < 0.01), which also had effectively reduced expression of intracellular HIF-1α mRNA and protein. Hence, SQBE had a protective effect on UVB-irradiated HaCaT cells and inhibited the UVB irradiation-induced expression of HIF-1α. This indicates that SQBE could prevent the occurrence of UVB radiation-induced skin cancer.
缺氧诱导因子-1α(HIF-1α)被认为是缺氧特异性细胞和发育反应的主要转录调节因子。本研究旨在探讨参芩生化提取物(SQBE)对紫外线B(UVB)照射的HaCaT细胞中HIF-1α表达的影响以及SQBE抗UVB诱导皮肤癌的可能作用机制。将对数生长期的HaCaT细胞接种于含10%胎牛血清的杜氏改良 Eagle培养基中,于37℃、5%CO₂条件下常规培养。细胞分为对照组(给予等量二甲基亚砜)、SQBE1组(12.5μg/mL SQBE)、SQBE2组(25.0μg/mL SQBE)和SQBE3组(50.0μg/mL SQBE)。给药4小时后,对照组和治疗组用UVB(0、20、40和60 mJ/cm)照射。24小时后,采用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)法检测细胞存活率。分别通过聚合酶链反应和蛋白质印迹法检测HIF-1α mRNA和蛋白的表达水平。经SQBE处理且UVB照射的细胞存活率有所提高。这种增加在SQBE3组最为显著(P<0.01),该组细胞内HIF-1α mRNA和蛋白的表达也有效降低。因此,SQBE对UVB照射的HaCaT细胞具有保护作用,并抑制UVB照射诱导的HIF-1α表达。这表明SQBE可预防UVB辐射诱导的皮肤癌的发生。