表皮生长因子调控核因子κB诱导卵巢上皮性癌紫杉醇耐药的作用及机制

目的探讨表皮生长因子(EGF)中上调的核因子κB(NF-κB)诱导卵巢上皮性癌(EOC)紫杉醇耐药的作用及机制。方法选取江苏省徐州市陆军第七十一集团军医院妇产科2015年1月至2019年1月收治的EOC亲本细胞系(A2780细胞)和紫杉醇耐药细胞系(A2780/Taxol细胞)及所对应的裸鼠实验分成A组(A2780组)、B组(A2780+CRM197组)、C组(A2780/Taxol组)、D(A2780/Taxol+CRM197组)四组,A组和C组加入100μg/ml DMSO培养基培养24 h,B组和D组加入100μg/ml CRM197培养基培养24 h。同时按NF-κB在紫杉醇的IC50及P-gp蛋白表达中的表达情况将上述分组细分成A1组(空载体)、B1组(NF-κB+siRNA组)、C1组(空载体+CRM197组)、D1组(NF-κB siRNA+CRM197组)。所有纳入者的肝素结合表皮生长因子(HB-EGF)和表皮生长因子受体(EGFR)表达均采用双重免疫荧光染色法检测,NF-κB表达采用蛋白印迹法检测,而裸鼠移植瘤组织(A2780/Taxol组与A2780/Taxol+CRM197组)中EGFR和P-糖蛋白(P-gp)表达则采用免疫组化SP法检测。结果C、D组HB-EGF、EGFR、NF-κB表达均低于A、B组,裸鼠移植瘤组织中NF-κB表达均低于A、B组(P<0.05),同时C组以上三项指标的表达及裸鼠移植瘤组织中NF-κB表达均高于A组(P<0.05),单NF-κB表达来看,EOC患者(A、B、C、D组)的NF-κB表达均低于裸鼠移植瘤组织中NF-κB表达(P<0.05)。B1、C1、D1组A2780/Taxol细胞对紫杉醇的IC50及P-gp蛋白表达均明显显著低于A1组,而B1、C1、D1组细胞内Rh123荧光强度则明显高于A1组(P<0.05);D1组A2780/Taxol细胞对紫杉醇IC50及P-gp蛋白表达均低于B1、C1组,Rh123荧光强度高于B1、C1组(P<0.05),其中B1与C1组的IC50、P-gp表达和Rh123荧光强度均无显著差异(P>0.05)。A2780/Taxol+CRM197组EGFR、P-gp表达强度均低于A2780/Taxol组(P<0.05)。结论在EOC的治疗中,EGF可通过调控EGFR、NF-κB、P-gp等信号通路而诱导其对紫杉醇产生耐药,NF-κB表达与EOC的紫杉醇耐药有相关性。

Effect and mechanism of epidermal growth factor on paclitaxel resistance induced by nuclear factor kappa B in ovarian epithelial cancer

Objective To investigate the effect and mechanism of up-regulation of nuclear factor kappa B(NF-kappa B)in epidermal growth factor(EGF)on paclitaxel resistance in epithelial ovarian cancer.Methods In the control experiment,the EOC parent cell line(A2780 cell line)and paclitaxel resistant cell line(A2780/taxol cell line)were admitted to the Department of gynecology and obstetrics of the 71 st Military Hospital of Xuzhou army from January 2015 to January 2019 and the corresponding nude mice were divided into four groups,A,B,C and D.Groups A and C were cultured with 100μg/ml DMSO medium for 24 h,and groups B and D were cultured with 100μg/ml CRM197 medium for 24 h.At the same time,the above groups were subdivided into four groups according to the expression of NF-κB in IC50 and P-gp protein of paclitaxel.The expression of HB-EGF and EGFR was detected by double immunofluorescence staining,NF-κB was detected by Western blotting,and EGFR,P-gp was detected by immunohistochemistry SP method in nude mice transplanted tumor tissues.Results The expressions of HB-EGF,EGFR and NF-κB in group C and D were lower than those in group A and B,and the expressions of NF-κB in nude mice transplanted tumors were lower than those in group A and B(P<0.05).At the same time,the expressions of the above three indicators in group C were higher than those in group A and group B(P<0.05).The expression of NF-κB in EOC patients(A,B,C and D groups)was lower than that in nude mice transplanted tumors(P<0.05).The expression of IC50 and P-gp protein in A2780/Taxol cells in B1,C1 and D1 group was significantly lower than that in A1 group,while the fluorescence intensity of Rh123 in A1 group was significantly highr than that in A1 group(P<0.05);the expression of IC50 and P-gp protein in D group was lower than that in B1 and C1 group,and the fluorescence intensity of Rh123 in B1 and C1 group was highr than that in B1 and C1 group(P<0.05).There was no significant difference in fluorescence intensity between the two groups(P>0.05).EGFR and P-gp expression in A2780/Taxol+CRM197 group were lower than those in A2780/Taxol group(P<0.05).Conclusion In the clinical treatment of EOC patients,EGF can induce resistance to paclitaxel by regulating EGFR,NF-κB,P-gp signaling pathways,and is related to NF-κB,that is,the expression of NF-κB is related to paclitaxel resistance of EOC.