RKIP对前列腺癌PC3细胞侵袭、转移的影响及机制
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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(qRTPCR).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.
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(qRTPCR).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.
引文
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[14]黄海,何旺,赖义明,等.PSMA激活PI3K/Akt通路对前列腺癌细胞增殖与凋亡及迁移能力的影响[J].中山大学学报:医学科学版,2016,37(4):490-496.
[15]贾艳菊,季晖.PI3K/Akt/mTOR信号传导通路与前列腺癌关系的研究进展[J].药学与临床研究,2015,23(2):160-163.
[16]BUFU T,DI X,YILIN Z,et al.Celastrol inhibits colorectal cancer cell proliferation and migration through suppression of MMP3and MMP7by the PI3K/Akt signaling pathway[J].Anticancer Drugs,2018,29(6):530-538.
[17]CHIU CT,CHEN JH,CHOU FP,et al.Hibiscus sabdariffa leaf extract inhibits human prostate cancer cell invasion via downregulation of Akt/NF-κB/MMP-9pathway[J].Nutrients,2015,7(7):5065-5087.
[18]LIU H,LI P,LI B,et al.RKIP promotes cisplatin-induced gastric cancer cell death through NF-κB/Snail pathway[J].Tumor Biol,2015,36(3):1445-1453.
[2]司富春,杜超飞.前列腺癌的中医证候和方药规律分析[J].中华中医药杂志,2015(2):581-585.
[3]谭书韬,吴斌.前列腺癌主动监测的研究进展[J].现代泌尿外科杂志,2016,21(1):68-72.
[4]YANG K,LI Y,LIAN G,et al.KRAS promotes tumor metastasis and chemoresistance by repressing RKIP via the MAPK-ERKpathway in pancreatic cancer[J].Int J Cancer,2018,142(11):2323-2334.
[5]MARTINHO O,FALOPPA CC,NETO CS,et al.Loss of RKIPexpression during the carcinogenic evolution of endometrial cancer[J].J Clin Pathol,2012,65(2):122-128.
[6]张璐,周亚敏,张凡,等.50岁以下年轻前列腺癌患者的临床特征及预后分析[J].现代泌尿外科杂志,2018,23(3):200-204.
[7]陈业辉,聂品,江文,等.前列腺癌骨转移的预测因素[J].南方医科大学学报,2016,36(2):205-209.
[8]HATZL S,GEIGER O,KUEPPER MK,et al.Increased expression of mir-23amediates a loss of expression in the RAF kinase inhibitor protein RKIP[J].Cancer Res,2016,76(12):3644-3654.
[9]蔡琴,王杰.Raf激酶抑制蛋白在恶性肿瘤中的研究进展[J].沈阳医学院学报,2016,18(4):284-286.
[10]张晓梅,谷欢,严璐,等.RKIP基因对胃癌细胞生物学行为的影响[J].中国现代医学杂志,2014,24(36):1-8.
[11]褚静,王立芹,曾娟.RKIP基因表达上调对宫颈癌细胞生物学行为影响[J].中国医药导刊,2016,18(8):833-834.
[12]HUANG Q,BAI F,NIE J,et al.Didymin ameliorates hepatic injury through inhibition of MAPK and NF-κB pathways by upregulating RKIP expression[J].Int Immunopharmacol,2017,42(1):130-138.
[13]FAES S,DORMOND O.PI3K and Akt:unfaithful partners in cancer[J].Int J Mol Sci,2015,16(9):21138-21152.
[14]黄海,何旺,赖义明,等.PSMA激活PI3K/Akt通路对前列腺癌细胞增殖与凋亡及迁移能力的影响[J].中山大学学报:医学科学版,2016,37(4):490-496.
[15]贾艳菊,季晖.PI3K/Akt/mTOR信号传导通路与前列腺癌关系的研究进展[J].药学与临床研究,2015,23(2):160-163.
[16]BUFU T,DI X,YILIN Z,et al.Celastrol inhibits colorectal cancer cell proliferation and migration through suppression of MMP3and MMP7by the PI3K/Akt signaling pathway[J].Anticancer Drugs,2018,29(6):530-538.
[17]CHIU CT,CHEN JH,CHOU FP,et al.Hibiscus sabdariffa leaf extract inhibits human prostate cancer cell invasion via downregulation of Akt/NF-κB/MMP-9pathway[J].Nutrients,2015,7(7):5065-5087.
[18]LIU H,LI P,LI B,et al.RKIP promotes cisplatin-induced gastric cancer cell death through NF-κB/Snail pathway[J].Tumor Biol,2015,36(3):1445-1453.