siRNA沉默PLK1基因对骨肉瘤细胞生物学特性影响
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摘要
目的探讨小分子干扰RNA(siRNA)沉默Polo样激酶1(PLK1)基因对骨肉瘤细胞生物学特性的影响。方法培养人骨肉瘤MG-63细胞,将其随机分为空白组、siRNA-对照序列组和siRNA-PLK1组。采用实时荧光定量PCR技术检测细胞中PLK1基因表达,Western blot法检测细胞中PLK1蛋白表达,CCK-8法检测细胞增殖活性,流式细胞术检测细胞凋亡情况,Transwell法检测细胞迁移和侵袭能力。结果 siRNA-PLK1组细胞中PLK1mRNA和蛋白相对表达量均低于空白组和siRNA-对照序列组,差异有统计学意义(F=129.125、62.363,P<0.01)。与空白组和siRNA-对照序列组相比,siRNA-PLK1组细胞培养48、72和96h时的增殖活性均降低,差异有统计学意义(F_(组间)=32.021,F_(时间)=357.249,F_(交互)=5.755,P<0.01)。与空白组和siRNA-对照序列组相比,siRNA-PLK1组细胞凋亡率升高,差异有统计学意义(F=69.863,P<0.01)。siRNA-PLK1组迁移细胞数和侵袭细胞数均低于空白组和siRNA-对照序列组,差异有统计学意义(F=28.998、34.783,P<0.01)。结论特异性沉默骨肉瘤细胞中PLK1基因表达可减少细胞增殖、增加细胞凋亡,抑制细胞侵袭、迁移能力。
Objective To investigate the effect of small interfering RNA(siRNA)silencing of polo-like kinase 1(PLK1)on the biological characteristics of osteosarcoma cells. Methods Human osteosarcoma MG-63 cells were cultured and randomly divided into blank group,siRNA-control sequence group,and siRNA-PLK1 group.Quantitative real-time PCR and Western blot were used to measure the mRNA and protein expression of PLK1 in cells;CCK-8 assay was used to measure cell proliferation acti-vity;flow cytometry was used to measure cell apoptosis;the Transwell method was used to measure cell migration and invasion abilities. Results The siRNA-PLK1 group had significantly lower relative mRNA and protein expression of PLK1 in cells than the blank group and the siRNA-control sequence group(F=129.125 and 62.363,P<0.01).Compared with the blank group and the siRNA-control sequence group,the siRNA-PLK1 group had significant reductions in proliferation activity at 48,72,and96 hof culture(F_(group)=32.021,F_(time)=357.249,F_(interaction)=5.755,P<0.01).Compared with the blank group and the siRNA-control sequence group,the siRNA-PLK1 group had a significant increase in cell apoptosis rate(F=69.863,P<0.01).The siRNAPLK1 group had significantly lower numbers of migrating cells and invasive cells than the control group and the siRNA-control sequence group(F=28.998 and 34.783,P<0.01). Conclusion Specific silencing of PLK1 gene expression in osteosarcoma cells can reduce cell proliferation,increase cell apoptosis,and inhibit cell invasion and migration.
Objective To investigate the effect of small interfering RNA(siRNA)silencing of polo-like kinase 1(PLK1)on the biological characteristics of osteosarcoma cells. Methods Human osteosarcoma MG-63 cells were cultured and randomly divided into blank group,siRNA-control sequence group,and siRNA-PLK1 group.Quantitative real-time PCR and Western blot were used to measure the mRNA and protein expression of PLK1 in cells;CCK-8 assay was used to measure cell proliferation acti-vity;flow cytometry was used to measure cell apoptosis;the Transwell method was used to measure cell migration and invasion abilities. Results The siRNA-PLK1 group had significantly lower relative mRNA and protein expression of PLK1 in cells than the blank group and the siRNA-control sequence group(F=129.125 and 62.363,P<0.01).Compared with the blank group and the siRNA-control sequence group,the siRNA-PLK1 group had significant reductions in proliferation activity at 48,72,and96 hof culture(F_(group)=32.021,F_(time)=357.249,F_(interaction)=5.755,P<0.01).Compared with the blank group and the siRNA-control sequence group,the siRNA-PLK1 group had a significant increase in cell apoptosis rate(F=69.863,P<0.01).The siRNAPLK1 group had significantly lower numbers of migrating cells and invasive cells than the control group and the siRNA-control sequence group(F=28.998 and 34.783,P<0.01). Conclusion Specific silencing of PLK1 gene expression in osteosarcoma cells can reduce cell proliferation,increase cell apoptosis,and inhibit cell invasion and migration.
引文
[1]WAN Jia,ZHANG Xianghong,LIU Tang,et al.Strategies and developments of immunotherapies in osteosarcoma[J].Oncology Letters,2016,11(1):511-520.
[2]CHEN Yu,XU Songfeng,XU Ming,et al.Postoperative infection and survival in osteosarcoma patients:reconsideration of immunotherapy for osteosarcoma[J].Mol Clin Oncol,2015,3(3):495-500.
[3]DAI Faxiang,XUAN Yi,JIN Jiejie,et al.CtBP2overexpression promotes tumor cell proliferation and invasion in gastric cancer and is associated with poor prognosis[J].Oncotarget,2017,8(17):28736-28749.
[4]KUMAR S,SHARMA A R,SHARMA G,et al.PLK-1:angel or devil for cell cycle progression[J].Biochimica et Biophysica Acta,2016,1865(2):190-203.
[5]LIU Zhixian,SUN Qingrong,WANG Xiaosheng.PLK1,a potential target for cancer therapy[J].Translational Oncology,2017,10(1):22-32.
[6]司晓辉,孙攀兴,杜增利.沉默WISP3基因抑制子宫内膜癌细胞增殖及侵袭力[J].齐鲁医学杂志,2017,32(6):631-635.
[7]余俊,刘彤鸥,李晓兰.小分子干扰RNA沉默黏着斑激酶基因对人宫颈癌Hela细胞生物学特征的影响[J].新乡医学院学报,2018,35(4):266-271.
[8]李国彬,张占成,王新颜.上调miR-200b对人喉癌Hep-2细胞增殖、迁移和侵袭能力的影响[J].山东大学耳鼻喉眼学报,2018,32(4):53-57.
[9]陈晓琦,蒋晶,陈欣菊,等.冬凌草甲素固体脂质纳米粒干预食管癌细胞的增殖[J].中国组织工程研究,2017,21(34):5460-5466.
[10]周宁,戴旖旎,白旭光,等.姜黄素对肝癌细胞凋亡及TNF-ɑ、IL-1β、IL-6炎性因子影响的机制研究[J].癌症进展,2017,15(11):1282-1285.
[11]李大刚,李辉宗,康乐.Vav3在小细胞肺癌组织中的表达及对细胞迁移和侵袭能力的影响[J].中国现代医学杂志,2018,28(24):32-37.
[12]罗玉政,陈波,李红樱.沉默IGF1R表达对胃癌细胞侵袭与迁移能力的影响及机制[J].中国临床研究,2018,31(2):163-166.
[13]任娟,徐叶峰,匡唐洪,等.104例骨肉瘤肺转移患者的预后及其影响因素[J].中华肿瘤杂志,2017,39(4):263-268.
[14]陈瑞玲,王刚阳,孙梦熊,等.骨肉瘤精准医学的实践与进展[J].中华实验外科杂志,2018,35(6):1190-1194.
[15]方三高,马强,杜娟,等.高级别表面骨肉瘤临床病理分析并文献复习[J].现代肿瘤医学,2018,26(9):1431-1436.
[16]KAMAL A F,WIDYAWARMAN H,HUSODO K,et al.Clinical outcome and survival of osteosarcomapatients in cipto mangunkusumo hospital:limb salvage surgery versus amputation[J].Acta Medica Indonesiana,2016,48(3):175-183.
[17]LU M H,FAN M F,YU X D.NSD2promotes osteosarcoma cell proliferation and metastasis by inhibiting E-cadherin expression[J].European Review for Medical and Pharmacological Sciences,2017,21(5):928-936.
[18]张景航,康小红,路平,等.长链非编码RNA MALAT1在骨肉瘤组织中的表达及其对骨肉瘤细胞侵袭和转移的影响[J].中华病理学杂志,2016,45(8):561-565.
[19]赫玮.Polo样蛋白激酶1参与有丝分裂调控的研究进展[J].医学综述,2015,21(20):3659-3661.
[20]周炳娟,马秋双,孙吉瑞,等.乳腺癌组织中FOXM1和PLK1的表达及意义[J].肿瘤防治研究,2017,44(3):193-196.
[21]WENGNG W T,SHIN J S,ROBERTS T L,et al.Molecular interactions of polo-like kinase 1in human cancers[J].Journal of Clinical Pathology,2016,69(7):557-562.
[22]陈阳,姚燕丹,罗曼莉,等.PLK1调控Pin1促进乳腺癌对他莫昔芬耐药的机制[J].热带医学杂志,2018,18(3):294-297.
[23]屈佳妮,甘润良,唐运莲.PLK1抑制剂Volasertib靶向治疗肿瘤的研究进展[J].中南医学科学杂志,2018,46(2):209-213.
[24]ZHAO C L,JU J Y,GAO W,et al.Downregulation of PLK1by RNAi attenuates the tumorigenicity of esophageal squamous cell carcinoma cells via promoting apoptosis and inhibiting angiogenesis[J].Neoplasma,2015,62(5):748-755.
[25]黄崇新,吕波,王跃,等.PLK1抑制剂对骨肉瘤细胞增殖的影响及其机制研究[J].骨科,2016,7(5):358-363.
[26]MAO Yonghuan,XI Ling,LI Quan,et al.Regulation of cell apoptosis and proliferation in pancreatic cancer through PI3K/Akt pathway via Polo-like kinase 1[J].Oncology Reports,2016,36(1):49-56.
[27]刘晓影,陈丽梅,张宝刚,等.慢病毒介导的人PLK1RNA干扰对食管鳞癌细胞裸鼠移植瘤的抑制作用及机制[J].中国药理学通报,2013,29(11):1528-1532.
[28]毛永欢,李泉,蔡则灵,等.Polo样激酶1对胰腺癌细胞株PANC-1凋亡、侵袭和迁移的影响[J].中华实验外科杂志,2016,33(11):2535-2537.
[29]于慧杰,吴晔,李峰生,等.敲低Plk1对人肝癌血管内皮细胞迁移影响研究[J].中华肿瘤防治杂志,2015,22(8):584-587.
[30]丁克云,徐娟,满昌峰,等.RNA干扰下调PLK1基因对恶性黑素瘤细胞侵袭和失巢凋亡的影响[J].中华皮肤科杂志,2014,47(6):413-416.
[2]CHEN Yu,XU Songfeng,XU Ming,et al.Postoperative infection and survival in osteosarcoma patients:reconsideration of immunotherapy for osteosarcoma[J].Mol Clin Oncol,2015,3(3):495-500.
[3]DAI Faxiang,XUAN Yi,JIN Jiejie,et al.CtBP2overexpression promotes tumor cell proliferation and invasion in gastric cancer and is associated with poor prognosis[J].Oncotarget,2017,8(17):28736-28749.
[4]KUMAR S,SHARMA A R,SHARMA G,et al.PLK-1:angel or devil for cell cycle progression[J].Biochimica et Biophysica Acta,2016,1865(2):190-203.
[5]LIU Zhixian,SUN Qingrong,WANG Xiaosheng.PLK1,a potential target for cancer therapy[J].Translational Oncology,2017,10(1):22-32.
[6]司晓辉,孙攀兴,杜增利.沉默WISP3基因抑制子宫内膜癌细胞增殖及侵袭力[J].齐鲁医学杂志,2017,32(6):631-635.
[7]余俊,刘彤鸥,李晓兰.小分子干扰RNA沉默黏着斑激酶基因对人宫颈癌Hela细胞生物学特征的影响[J].新乡医学院学报,2018,35(4):266-271.
[8]李国彬,张占成,王新颜.上调miR-200b对人喉癌Hep-2细胞增殖、迁移和侵袭能力的影响[J].山东大学耳鼻喉眼学报,2018,32(4):53-57.
[9]陈晓琦,蒋晶,陈欣菊,等.冬凌草甲素固体脂质纳米粒干预食管癌细胞的增殖[J].中国组织工程研究,2017,21(34):5460-5466.
[10]周宁,戴旖旎,白旭光,等.姜黄素对肝癌细胞凋亡及TNF-ɑ、IL-1β、IL-6炎性因子影响的机制研究[J].癌症进展,2017,15(11):1282-1285.
[11]李大刚,李辉宗,康乐.Vav3在小细胞肺癌组织中的表达及对细胞迁移和侵袭能力的影响[J].中国现代医学杂志,2018,28(24):32-37.
[12]罗玉政,陈波,李红樱.沉默IGF1R表达对胃癌细胞侵袭与迁移能力的影响及机制[J].中国临床研究,2018,31(2):163-166.
[13]任娟,徐叶峰,匡唐洪,等.104例骨肉瘤肺转移患者的预后及其影响因素[J].中华肿瘤杂志,2017,39(4):263-268.
[14]陈瑞玲,王刚阳,孙梦熊,等.骨肉瘤精准医学的实践与进展[J].中华实验外科杂志,2018,35(6):1190-1194.
[15]方三高,马强,杜娟,等.高级别表面骨肉瘤临床病理分析并文献复习[J].现代肿瘤医学,2018,26(9):1431-1436.
[16]KAMAL A F,WIDYAWARMAN H,HUSODO K,et al.Clinical outcome and survival of osteosarcomapatients in cipto mangunkusumo hospital:limb salvage surgery versus amputation[J].Acta Medica Indonesiana,2016,48(3):175-183.
[17]LU M H,FAN M F,YU X D.NSD2promotes osteosarcoma cell proliferation and metastasis by inhibiting E-cadherin expression[J].European Review for Medical and Pharmacological Sciences,2017,21(5):928-936.
[18]张景航,康小红,路平,等.长链非编码RNA MALAT1在骨肉瘤组织中的表达及其对骨肉瘤细胞侵袭和转移的影响[J].中华病理学杂志,2016,45(8):561-565.
[19]赫玮.Polo样蛋白激酶1参与有丝分裂调控的研究进展[J].医学综述,2015,21(20):3659-3661.
[20]周炳娟,马秋双,孙吉瑞,等.乳腺癌组织中FOXM1和PLK1的表达及意义[J].肿瘤防治研究,2017,44(3):193-196.
[21]WENGNG W T,SHIN J S,ROBERTS T L,et al.Molecular interactions of polo-like kinase 1in human cancers[J].Journal of Clinical Pathology,2016,69(7):557-562.
[22]陈阳,姚燕丹,罗曼莉,等.PLK1调控Pin1促进乳腺癌对他莫昔芬耐药的机制[J].热带医学杂志,2018,18(3):294-297.
[23]屈佳妮,甘润良,唐运莲.PLK1抑制剂Volasertib靶向治疗肿瘤的研究进展[J].中南医学科学杂志,2018,46(2):209-213.
[24]ZHAO C L,JU J Y,GAO W,et al.Downregulation of PLK1by RNAi attenuates the tumorigenicity of esophageal squamous cell carcinoma cells via promoting apoptosis and inhibiting angiogenesis[J].Neoplasma,2015,62(5):748-755.
[25]黄崇新,吕波,王跃,等.PLK1抑制剂对骨肉瘤细胞增殖的影响及其机制研究[J].骨科,2016,7(5):358-363.
[26]MAO Yonghuan,XI Ling,LI Quan,et al.Regulation of cell apoptosis and proliferation in pancreatic cancer through PI3K/Akt pathway via Polo-like kinase 1[J].Oncology Reports,2016,36(1):49-56.
[27]刘晓影,陈丽梅,张宝刚,等.慢病毒介导的人PLK1RNA干扰对食管鳞癌细胞裸鼠移植瘤的抑制作用及机制[J].中国药理学通报,2013,29(11):1528-1532.
[28]毛永欢,李泉,蔡则灵,等.Polo样激酶1对胰腺癌细胞株PANC-1凋亡、侵袭和迁移的影响[J].中华实验外科杂志,2016,33(11):2535-2537.
[29]于慧杰,吴晔,李峰生,等.敲低Plk1对人肝癌血管内皮细胞迁移影响研究[J].中华肿瘤防治杂志,2015,22(8):584-587.
[30]丁克云,徐娟,满昌峰,等.RNA干扰下调PLK1基因对恶性黑素瘤细胞侵袭和失巢凋亡的影响[J].中华皮肤科杂志,2014,47(6):413-416.