鼻咽癌组织中WNT8B与ZNF191蛋白表达与临床病理及预后的相关性分析

目的 探讨鼻咽癌组织中Wnt家族成员8B(Wnt family member 8B,WNT8B)与锌指转录因子191(zinc finger transcription factor 191,ZNF191)蛋白表达与临床病理及预后的相关性。方法 收集2017年1月~2019年1月东阳市人民医院保存的126例鼻咽癌组织石蜡标本及其临床病理资料(病例组),另选取慢性鼻咽炎组织样本40例(对照组)。采用免疫组织化学法检测2组样本组织中WNT8B和ZNF191蛋白表达情况,并对比两蛋白阳性率及共表达阳性率,分析病例组患者鼻咽癌组织中WNT8B和ZNF191蛋白表达与临床病理因素关系,采用Kaplan-Meier法和Cox回归分析法分析鼻咽癌组织中WNT8B和ZNF191表达与预后的关系。结果 病例组鼻咽癌组织中WNT8B和ZNF191阳性率及二者共表达阳性率均高于对照组鼻咽黏膜组织(P<0.05)。Spearson相关性分析结果显示,鼻咽癌组织中WNT8B与ZNF191表达呈正相关(P<0.05)。T3~T4级、Ⅲ~Ⅳ期、有淋巴结转移的鼻咽癌组织中WNT8B和ZNF191阳性率及二者共表达阳性率分别高于T1~T2级、Ⅰ~Ⅱ期、无淋巴结转移患者(P<0.05)。WNT8B阳性与阴性患者3年生存率分别为57.89%、83.87%,ZNF191阳性与阴性患者分别为56.79%、77.78%,共表达阳性患者与非共表达患者分别为50.75%、79.66%;经Kaplan-Meier分析结果显示,WNT8B阳性患者与阴性、ZNF191阳性与阴性、共表达阳性与非共表达患者累积存活曲线比较,差异均有统计学意义(P<0.05)。Cox回归分析显示,浸润深度[HR(95%CI)=1.980(1.023~3.955)]、TNM分期[HR(95%CI)=3.207(1.936~11.751)]、WNT8B阳性表达[HR(95%CI)=4.506(2.068~13.774)]、ZNF191阳性表达[HR(95%CI)=3.114(2.061~10.485)]、WNT8B+ZNF191共表达[HR(95%CI)=4.691(2.144~14.902)]是影响总生存期的独立危险因素(P<0.05)。结论 鼻咽癌患者鼻咽癌组织中WNT8B和ZNF191阳性表达率高,WNT8B和ZNF191表达与鼻咽癌患者浸润深度、TNM分期、淋巴结转移及预后生存相关。

Expression and function of WNT4 involved in larvae development and limb regeneration in Portunus trituberculatus

The wingless-related integration site(WNT)proteins are a family of secreted glycoproteins that are evolutionarily conserved and are believed to be involved in evolution in vertebrates and invertebrates.WNT signaling pathways may be associated with limb regeneration and development in crustaceans.However,the detail mechanisms remain unclear.Therefore,the distribution of WNT4 in the hepatopancreas,muscle,hemocyte,ganglion,heart,eyestalk,gill tissue,and diff erent larvae development stages of the swimming crab(Portunus trituberculatus)were characterized using immunofl uorescence,real-time PCR,and Western blotting.Signifi cant PtWNT4 expression was detected in heart and eyestalk.In addition,PtWNT4 was expressed in all larval stages of P.trituberculatus with a dynamic expression pattern,especially in the eyestalk and other organs in the carapace area.The injection of WNT4 dsRNA into regenerative limbs signifi cantly decreased PtWNT4 mRNA levels in the eyestalk,heart,and muscle,resulting in 1.9-fold,2.2-fold,and 2.7-fold decreases compared with those detected in the group injected with crab saline(P<0.05),respectively,indicating successful gene silencing.Overall,expression analysis on the WNT4 using RNAi provides an insight to its functional mechanism during limb regeneration in P.trituberculatus.The results not only demonstrated the requirement for WNT4 in limb regeneration of Crustaceans,but also suggested its ability to promote larval development at specifi c stages.

WNT5A drives interleukin-6-dependent epithelial-mesenchymal transition via the JAK/STAT pathway in keloid pathogenesis

Background:Keloid scarring is a fibroproliferative disease caused by aberrant genetic activation with an unclear underlying mechanism.Genetic predisposition,aberrant cellular responses to environmental factors,increased inflammatory cytokines and epithelial-mesenchymal transition(EMT)phenomena are known as major contributors.In this study,we aimed to identify the molecular drivers that initiate keloid pathogenesis.Methods:Bulk tissue RNA sequencing analyses of keloid and normal tissues along with ex vivo and in vitro tests were performed to identify the contributing genes to keloid pathogenesis.An animal model of inflammatory keloid scarring was reproduced by replication of a skin fibrosis model with intradermal bleomycin injection in C57BL/6 mice.Results:Gene set enrichment analysis revealed upregulation of Wnt family member 5A(WNT5A)expression and genes associated with EMT in keloid tissues.Consistently,human keloid tissues and the bleomycin-induced skin fibrosis animal model showed significantly increased expression ofWNT5A and EMT markers.Increased activation of the interleukin(IL)-6/Janus kinase(JAK)/signal transducer and activator of transcription(STAT)pathway and subsequent elevation of EMT markerswas also observed in keratinocytes co-cultured withWNT5A-activated fibroblasts or keloid fibroblasts.Furthermore,WNT5A silencing and the blockage of IL-6 secretion via neutralizing IL-6 antibody reversed hyperactivation of the STAT pathway and EMT markers in keratinocytes.Lastly,STAT3 silencing significantly reduced the EMT-like phenotypes in both keratinocytes and IL-6-stimulated keratinocytes.Conclusions:Intercellular communication via the WNT5A and STAT pathways possibly underlies a partial mechanism of EMT-like phenomena in keloid pathogenesis.IL-6 secreted from WNT5A-activated fibroblasts or keloid fibroblasts activates the JAK/STAT signaling pathway in adjacent keratinocytes which in turn express EMT markers.A better understanding of keloid development and the role of WNT5A in EMT will promote the development of next-generation targeted treatments for keloid scars.

Wnt8a is one of the candidate genes that play essential roles in the elongation of the seahorse prehensile tail

Seahorses are a hallmark of specialized morphological features due to their elongated prehensile tail.However,the underlying genomic grounds of seahorse tail development remain elusive.Herein,we evaluated the roles of essential genes from the Wnt gene family for the tail developmental process in the lined seahorse(Hippocampus erectus).Comparative genomic analysis revealed that the Wnt gene family is conserved in seahorses.The expression profiles and in situ hybridization suggested that Wnt5a,Wnt8a,and Wnt11 may participate in seahorse tail development.Like in other teleosts,Wnt5a and Wntll were found to regulate the development of the tail axial mesoderm and tail somitic mesoderm,respectively.However,a significantly extended expression period of Wnt8a during seahorse tail development was observed.Signaling pathway analysis further showed that Wnt5a up-regulated the expression of the tail axial mesoderm gene(Shh),while interaction analysis indicated that Wnt8a could promote the expression of Wntll.In summary,our results indicate that the special extended expression period of Wnt8a might promote caudal tail axis formation,which contributes to the formation of the elongated tail of the seahorse.