摘要
目的探究Raf激酶抑制蛋白(RKIP)过表达对前列腺癌PC3细胞侵袭、转移及磷脂酰肌醇激酶(PI3K)/苏氨酸蛋白激酶(Akt)/核转录因子(NF-kB)通路的影响。方法蛋白免疫印迹(Western blot)检测前列腺癌肿瘤组织、癌旁组织中RKIP蛋白表达情况。将pcDNA3.0-RKIP、pcDNA3.0、RKIP-shRNA和shRNA-NC转染至前列腺癌细胞,命名为pcDNA3.0-RKIP组、pcDNA3.0组、RKIP-shRNA组、shRNA-NC组,以未经转染的PC3细胞作为对照组,采用实时荧光定量PCR(qRT-PCR)检测PC3中RKIP mRNA表达情况,分别采用transwell小室和划痕实验检测PC3细胞侵袭和转移能力,蛋白Western blot检测PC3细胞中PI3K、Akt、及NF-kB表达情况。结果前列腺癌肿瘤组织RKIP蛋白表达水平显著低于癌旁组织(P<0.05);pcDNA3.0-RKIP组PC3细胞中RKIP mRNA表达水平显著高于RKIP-shRNA组和对照组(P<0.05);pcDNA3.0-RKIP组PC3细胞侵袭和迁移能力显著低于RKIP-shRNA组和对照组(P<0.05);pcDNA3.0-RKIP组PC3细胞中PI3K、p-Akt和NF-kB的表达水平显著低于RKIP-shRNA组和对照组(P<0.05)。结论 RKIP过表达能够抑制前列腺癌细胞侵袭和转移,并能够抑制PI3K/Akt/NF-κB通路相关蛋白的表达,为前列腺癌的靶向治疗提供一定的理论依据。
Objective To investigate the effects of overexpression of Raf kinase inhibitor(RKIP)on the invasion,metastasis and phosphatidylinositol kinase(PI3K)/threonine protein kinase(Akt)/nuclear transcription factor(NF-kB)pathway in prostate cancer PC3 cells.Methods The protein expressions of RKIP in prostate cancer and paracancerous tissues were detected with Western blot.PC3 cells were transfected with pcDNA3.0-RKIP,pcDNA3.0,RKIP-shRNA and shRNA-NC,and divided into pcDNA3.0-RKIP group,pcDNA3.0 group,RKIP-shRNA group and shRNA-NC group,and non-transfected PC3 cells served as the control group.The mRNA expressions of RKIP were detected with real-time fluorescence quantitative PCR(qRT-PCR).The invasion and metastasis of PC3 cells were assessed with Transwell assay and scratch test.The expressions of PI3K,Akt,p-Akt and NF-kB were determined with Western blot.Results The protein expression of RKIP in cancer tissues was significantly lower than that in paracancerous tissues(P<0.05).The mRNA expression of RKIP in pcDNA3.0-RKIP group was significantly higher than that in RKIP-shRNA group and control group(P<0.05).The invasion and migration of pcDNA3.0-RKIP group were significantly lower than those in RKIP-shRNA group and control group(P<0.05).The expressions of PI3K,p-Akt and NF-kB in pcDNA3.0-RKIP group were lower than those in the RKIP-shRNA group and control group(P<0.05).Conclusion Overexpression of RKIP can inhibit the invasion and metastasis of prostate cancer cells,and inhibit the expression of PI3K/Akt/NF-kB pathway-related proteins.Therefore,it can provide a theoretical basis for the targeted treatment of prostate cancer.
作者
赵鹏程
杨栋
任乐
何朝宏
ZHAO Peng-cheng;YANG Dong;REN Le;HE Chao-hong(Department of Urology,Henan Cancer Hospital,Affiliated Cancer Hospital of Zhengzhou University,Henan 450003,China)
出处
《现代泌尿外科杂志》
CAS
2018年第12期956-960,974,共6页
Journal of Modern Urology
