1.Inhibition of melanoma cell proliferation by targeting Wnt/β-catenin pathway through Sox4 RNA interference.
Huahua, CAI ; Anhong, NI ; Wen, LI ; Jiawen, LI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2011;31(4):565-9
The effect of siRNA-mediated Sox4 gene silencing on Wnt/β-catenin signaling pathway of human malignant melanoma cell line A375 was investigated. Two types of dsRNA targeting Sox4 were constructed and transfected into A375 cells, and untreated cells and cells transfected with scramble RNA were used as blank control and negative control respectively. The expression levels of mRNA and protein of Sox4, Wnt3a, β-catenin and Wnt/β-catenin signaling target gene Survivin were detected after real-time PCR and Western blot respectively. MTT assay was used to measure cell proliferation after Sox4 knockdown. β-catenin/TCF transcription reporter assay was used for assessing Wnt/β-catenin signaling pathway activity. Our results showed that the two types of Sox4 siRNA were transfected into A375 cells successfully. As compared with untreated cells, Sox4 siRNAs had no significant influence on Wnt3a expression, and Sox4 siRNAs led to the decrease of β-catenin at protein level. Wnt/β-catenin signaling pathway activity was inhibited significantly. As a target of Wnt/β-catenin signaling, Survivin was decreased at both mRNA and protein levels, and cell proliferation was attenuated. Our study suggests that Sox4 may play an important role in Wnt/β-catenin signaling pathway in human malignant melanoma cells by regulating β-catenin protein level, indicating that Sox4 is involved in the progression of malignant melanoma through Wnt/β-catenin signaling pathway.
2.Expression of IRF-4 and IBP in skin lesions of patients with psoriasis vulgaris.
Anhong, NI ; Hongxiang, CHEN ; Yan, WU ; Wen, LI ; Sh, ANJUAN CHEN ; Jiawen, LI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2012;32(2):287-90
The expression of the interferon regulatory factor 4 (IRF-4) and the IRF-4-binding protein (IBP) in psoriatic skin lesions was investigated. The expression of IRF-4 and IBP in skin lesions of 20 patients with psoriasis vulgaris were immunohistochemically dectected. Normal skin from 10 healthy people was used as normal control. The study showed that expression of IRF-4 was increased significantly in keratinocytes and inflammatory cells in the lesions of psoriasis vulgaris than that in the normal control. The detection revealed that IBP expression in keratinocytes, lymphocytes, hair follicles, and sebaceous glands in normal skin was significantly lower than that in the lesions of psoriasis vulgaris (P<0.05). Both IRF-4 and IBP might be involved in the pathogenesis of psoriasis vulgaris.
3.Inhibition of Melanoma Cell Proliferation by Targeting Wnt/β-catenin Pathway through Sox4 RNA Interference
CAI HUAHUA ; NI ANHONG ; LI WEN ; LI JIAWEN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2011;31(4):565-569
The effect of siRNA-mediated Sox4 gene silencing on Wnt/β-catenin signaling pathway of human malignant melanoma cell line A375 was investigated.Two types of dsRNA targeting Sox4 were constructed and transfected into A375 cells,and untreated cells and cells transfected with scramble RNA were used as blank control and negative control respectively.The expression levels of mRNA and protein of Sox4,Wnt3a,β-catenin and Wnt/β-catenin signaling target gene Survivin were detected after real-time PCR and Western blot respectively.MTT assay was used to measure cell proliferation after Sox4 knockdown.β-catenin/TCF transcription reporter assay was used for assessing Wnt/β-catenin signaling pathway activity.Our results showed that the two types of Sox4 siRNA were transfected into A375 cells successfully.As compared with untreated cells,Sox4 siRNAs had no significant influence on Wnt3a expression,and Sox4 siRNAs led to the decrease of β-catenin at protein level.Wnt/β-catenin signaling pathway activity was inhibited significantly.As a target of Wnt/β-catenin signaling,Survivin was decreased at both mRNA and protein levels,and cell proliferation was attenuated.Our study suggests that Sox4 may play an important role in Wnt/β-catenin signaling pathway in human malignant melanoma cells by regulating β-catenin protein level,indicating that Sox4 is involved in the progression of malignant melanoma thromgh Wnt/β-catenin signaling pathway.
4.Expression of IRF-4 and IBP in Skin Lesions of Patients with Psoriasis Vulgaris
NI ANHONG ; CHEN HONGXIANG ; WU YAN ; LI WEN ; CHEN SHANJUAN ; LI JIAWEN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2012;32(2):287-290
The expression of the interferon regulatory factor 4 (IRF-4) and the IRF-4-binding protein (IBP) in psoriatic skin lesions was investigated.The expression of IRF-4 and IBP in skin lesions of 20 patients with psoriasis vulgaris were immunohistochemically dectected.Normal skin from 10 healthy people was used as normal control.The study showed that expression of IRF-4 was increased significantly in keratinocytes and inflammatory cells in the lesions of psoriasis vulgaris than that in the normal control.The detection revealed that IBP expression in keratinocytes,lymphocytes,hair follicles,and sebaceous glands in normal skin was significantly lower than that in the lesions of psoriasis vulgaris (P<0.05).Both IRF-4 and IBP might be involved in the pathogenesis of psoriasis vulgaris.