摘要
目的 YAP作为Hippo信号通路的主要蛋白,调控多种肿瘤细胞的增殖。本研究探讨Hippo/YAP信号通路对HPV感染宫颈癌细胞TGF-α及EGFR表达的调节作用。方法免疫组化法检测HPV-16(+)Ca Ski细胞、HPV-18(+)C4-1细胞,HPV(-)人宫颈癌细胞C33a中YAP、TGF-α及EGFR的表达。给予YAP抑制剂Vertepofin干预后,CCK-8法检测对Ca Ski及C4-1细胞增殖影响,流式细胞术检测细胞周期,蛋白质印迹法及Real time PCR法检测YAP、TGF-α、EGFR蛋白及mRNA的表达。结果 Ca Ski、C4-1细胞YAP、TGF-α、EGFR蛋白水平均高于HPV(-)组C33a细胞,差异有统计学意义,均P<0.001。给予YAP抑制剂Vertepofin后,在Ca Ski细胞中,干预后第8天减缓速度更加明显,P=0.005;在C4-1细胞中,在干预后第8天后,增殖速度较正常培养减缓明显,P=0.004。干预后CaSki及C4-1细胞处于G0-G1期,且差异有统计学意义,F值分别为463.207和65.872,均P<0.01。在抑制YAP表达的同时,TGF-α、EGFR的mRNA及蛋白随之下降。结论 Hippo/YAP信号通路对HPV感染宫颈癌细胞中TGF-α与EGFR的表达具有正调控作用。
OBJECTIVE YAP as the major protein of Hippo signaling pathway,regulates the proliferation of various tumor cells.In this study,we investigated the role of Hippo/YAP signaling in the regulation of TGF-a and EGFR expression in cervical cancer cells infected by HPV.METHODS The expressions of YAP,TGF-a and EGFR in HPV-16(+), HPV-18(+) human cervical cancer cells Ca Ski,C4-1 and HPV(-) human cervical cancer cells C33a were detected by immunohistochemistry.The effect of YAP inhibitor Vertepofin on the proliferation of Ca Ski and C4-1 cells was detected by CCK-8 assay.Flow cytometry was used to detect cell cycle.The expression of YAP.TGF-a,EGFR protein and mRNA were detected by Western blotting and Realtime PCR.RESULTS The protein levels of YAP,TGF-a and EGFR in Ca Ski and C4-1 cells were higher than those in HPV(-) group and the P values were all<0.001 ,and the difference was statistically significant.After the YAP inhibitor Vertepofin was administered?the rate of mitigation was more pronounced in the Ca Ski cells on the 8th day after the intervention (P=0.005).In the C4-1 cells,the proliferation rate was slower than the n ormal culture on the 8th day after the intervention (P=0.004).CaSki and C4-1 cells were in G0(ri phase after in tervention, and the difference was statistically significant (F=463.207,P=0.00;F=65.872,P=0.01).While suppressing the expression of YAP,the mRNA and protein of TGF-a and EGFR decreased.CONCLUSION The Hippo/YAP signaling pathway has a positive regulatory effect on the expression of TGF-a and EGFR in cervical cancer cells infected by HPV.
作者
裴峰
罗倩
吕文静
PEI Feng;LUO Qian;LU Wen-jing(Department of Laboratory First People's Hospital of Tianmen,Tianmen 431700,P.R.China)
出处
《中华肿瘤防治杂志》
CAS
北大核心
2019年第3期148-154,共7页
Chinese Journal of Cancer Prevention and Treatment
基金
山东省中医药科技发展计划(2017-279)
该项目于2017年9月25日在山东立项
