Contribution of Decreased Expression of Ku70 to Enhanced Radiosensitivity by Sodium Butyrate in Glioblastoma Cell Line（U251）

The present study investigated the enhanced radiosensitivity of U-251 cells induced by sodium butyrate(NaB) and its possible mechanisms.Increased radiosensitivity of U251 cells was examined by clonogenic cell survival assays.The expression of Ku70 mRNA and protein was detected by using RT-PCR and Western blotting respectively.γ-H2AX foci were measured at different time points after ionizing irradiation alone or combined with NaB treatment.The results showed that cell survival rate was significantly reduced,both D0 and Dq values were decreased(D0:1.43 Gy vs.1.76 Gy;Dq:1.22 Gy vs.2.05 Gy) after the combined treatment as compared with irradiation alone,and sensitivity enhancing ratio(SER) reached 1.23.The average number of γ-H2AX foci per cell receiving the combined treatment was significantly increased at different time points,and the expression levels of Ku70 mRNA and protein were suppressed by NaB in a dose-dependent manner.It was concluded that enhanced radiosensitivity induced by NaB involves an inhibited expression of Ku70 and an increase in γ-H2AX foci,which suggests decreased ability in DSB repair.     

Effect of prolonging interval time between coronary angiography and percutaneous coronary intervention on X-ray-induced DNA double-strand breaks in blood lymphocytes

Background It is desirable to minimize the risk of adverse radiation effects associated with percutaneous coronary intervention.The aim of this study was to determine the impact of prolonging the interval between coronary angiography and percutaneous coronary intervention on X-ray-induced DNA double-strand breaks in blood lymphocytes using γ-H2AX immunofluorescence microscopy.Methods Blood samples of eight patients were taken before the first exposure to ionizing radiation,10 minutes,20 minutes,30 minutes,1 hour,and 24 hours after the last exposure to determine the γ-H2AX foci repair kinetics.Fifty-eight patients undergoing percutaneous coronary intervention were randomized to an intermittent radiation exposure group and a continuous radiation exposure group.Blood samples were taken before coronary angiography and 15 minutes after the last exposure.By enumerating γ-H2AX foci,the impact of prolonging the interval on DNA double-strand breaks was investigated.Student t-test was used to compare the difference in DNA double-strand breaks between the two groups.Results An increase in foci was found in all patients received percutaneous coronary intervention.The maximum number of γ-H2AX foci was found 10-20 minutes after the end of the last exposure.There was no statistically significant difference between the two groups in γ-H2AX foci at baseline.On average there were （0.79±0.15） γ-H2AX foci induced by interventional X-rays per lymphocyte in the continuous radiation exposure group and （0.66±0.21） in the intermittent radiation exposure group after exposure （P〈0.05）.Conclusions A significant number of γ-H2AX foci develop following the percutaneous coronary intervention procedures.The number of X-ray-induced DNA double-strand breaks may be decreased by prolonging the interval time between coronary angiography and percutaneous coronary intervention to 30 minutes.     

Mitogen-activated protein kinase-activated protein kinase 2 regulates tumor necrosis factor-induced interleukin-6 expression via human antigen R

Background Human antigen R （HuR） is a ubiquitously expressed member of the ELAV family, and has relatively high cytoplasmic abundance in lung tissue regenerating after injury. In this study, we investigated whether mitogen-activated protein kinase （MAPK）-activated protein kinase 2 （MK2） and HuR participate in the tumor necrosis factor （TNF）-induced expression of interleukin-6 （IL-6）. Methods Human pulmonary microvascular endothelial cells were treated with TNF following short interfering RNAmediated knockdown of MK2 or HuR. Cell supernatants were collected to detect the mRNA and protein expression of IL-6 at different time points, The expression and half-life of IL-6 mRNA were then determined in cells that had been treated with actinomycin D. Finally, after knockdown of MK2, the cytoplasmic expression of HuR protein was analyzed using Western blotting. Results MK2 or HuR knockdown decreased both the mRNA and protein expression of IL-6 in TNF-stimulated cells. In MK2 knockdown cells, the half-life of IL-6 mRNA was reduced to 36 minutes, compared with 67 minutes in the control group. In HuR knockdown cells, the half-life of IL-6 mRNA decreased from 62 minutes to 24 minutes. Further analysis revealed that knockdown of MK2 resulted in reduced HuR protein expression in the cytoplasm. Conclusions MK2 regulates the TNF-induced expression of IL-6 by influencing the cytoplasmic levels of HuR.     

Cigarette smoke extract promotes human pulmonary artery smooth muscle cells proliferation through protein kinase C alpha-dependent induction of cyclin D1

Background Exposure to cigarette smoke stimulates the proliferation of human pulmonary artery smooth muscle cells （HPASMCs） in vivo and in vitro. However, the molecular mechanism remains unclear. This study aimed at investigating the role of signaling pathways involving protein kinase C alpha （PKCα） and cyclin D1 in the cigarette smoke extract （CSE）-induced HPASMCs proliferation.Methods Synchronized HPASMCs were treated with different concentrations of CSE. Cell proliferation was evaluated by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyttetrazolium bromide （MTT） assay and cell counting. Cell cycle was analyzed by flow cytometry with propidium iodide staining. Activation of PKCα was measured by detecting the expression of PKCαprotein in the cytosolic and membrane fractions using Western blotting analysis. Small interfering RNA （siRNA） was used to knockdown PKCα and cyclin D1. The cyclin D1 mRNA was assessed by real-time RT-PCR. The PKCα and cyclin D1protein levels were detected by Western blotting.Results Low concentrations of CSE （1%-10%） stimulated proliferation of HPASMCs, with its maximal effect at 5%.CSE （5%） led to PKCα activation. Inhibition of PKCα activity using G（o） 6976 or siRNA-mediated knockdown of PKCα significantly attenuated CSE-induced cell proliferation and G1/S transition. Cyclin D1, one of key regulators of G1/S transition, was found to be upregulated by 5% CSE at both the mRNA and protein levels. CSE-stimulated cellproliferation and G1/S transition was abolished by cyclin D1 siRNA. Moreover, G（o） 6976 or PKCα siRNA significantlysuppressed CSE-induced upregulation of cyclin D1 at both the mRNA and protein levels.Conclusion PKCα-cyclin D1 pathway at least partially mediates the CSE-induced proliferation in HPASMCs.     

Vascular Endothelial Growth Factor165-regulated Nasopharyngeal Carcinoma Cell Lines Invasion and Migration Involve Expression and Activation of Matrix Metalloproteinase-2

The effect of vascular endothelial growth factor （VEGF） overexpression on matrix metalloproteinase-2 （MMP-2） in nasopharyngeal carcinoma （NPC） cells in vitro and the possible mechanism involved were investigated, and the correlation between the expression of VEGF and MMP-2 in NPC evaluated. The NPC cells were transfected with PAd-trackVEGF165 plasmid. The expression levels of VEGF and MMP-2 mRNA and protein in NPC cells were detected by semi-quantitative RT-PCR and Western blot respectively. It was found that the expression of VEGF and MMP-2 mRNA and protein was significantly increased in NPC cells after transfection of VEGF 165. It was concluded that the expression of VEGF was correlated to the in vitro invasion of NPC cells, and the induction of MMP-2 by VEGF was a key process of NPC cell invasion.     

Vascular Endothelial Growth Factorl65-regulated Nasopharyngeal Carcinoma Cell Lines Invasion and Migration Involve Expression and Activation of Matrix Metalloproteinase-2

The effect of vascular endothelial growth factor (VEGF) overexpression on matrix met-alloproteinase-2 (MMP-2) in nasopharyngeal carcinoma (NPC) cells in vitro and the possible mechanism involved were investigated, and the correlation between the expression of VEGF and MMP-2 in NPC evaluated. The NPC cells were transfected with PAd-trackVEGF165 plasmid. The expression levels of VEGF and MMP-2 mRNA and protein in NPC cells were detected by semi-quantitative RT-PCR and Western blot respectively. It was found that the expression of VEGF and MMP-2 mRNA and protein was significantly increased in NPC cells after transfection of VEGF165. It was concluded that the expression of VEGF was correlated to the in vitro invasion of NPC cells, and the induction of MMP-2 by VEGF was a key process of NPC cell invasion.     

Growth and activation of PI-3K/PKB and Akt by stromal cell-derived factor 1α in endometrial carcinoma cells with expression of suppressor endoprotein PTEN

Background Mutation or deletion in the phosphatase and tensin homologue deleted on chromosome ten （PTEN） gene has been identified as an important cause of endometrial carcinoma; stromal cell derived factor-1α （SDF-1α） exerts growth-promoting effects on endometrial cancer cells through activation of the PI-3 kinase/Akt pathway and downstream effectors such as extracellular-responsive kinase （ERK）. In this study, a plasmid containing the PTEN gene was transfected into Ishikawa cells to investigate the difference in growth and signal transduction between Ishikawa-PTEN and Ishikawa cells after SDF-1α stimulation, and to study mechanisms of the involvement of PTEN protein in endometrial carcinoma development. Methods Ishikawa cells were transfected with a plasmid （pLXSN-PTEN） containing the PTEN gene and a plasmid （pLXSN-EGFP） with enhanced green fluorescent protein （EGFP）. Cells were then screened to obtain Ishikawa-PTEN cells and Ishikawa-neo cells that can both stably express PTEN protein and EGFP. Expression of PTEN protein, phosphorylation levels of AKT and ERK （pAKT and pERK） and growth differences in Ishikawa-PTEN, Ishikawa-neo and Ishikawa cells before and after SDF-1α stimulation were then determined by Western blots and MTT assays. Results Western blot analysis showed that Ishikawa cells produced PTEN after transfection with the PTEN gene. At 15 minutes after SDF-1α stimulation, the pAKT level of Ishikawa-PTEN cells was lower than that of Ishikawa-neo cells and Ishikawa cells. There was no significant difference in pERK levels among the three cell lines. The positive effect of SDF-1α on Ishikawa-PTEN cells growth was markedly less than the effect on Ishikawa-neo and Ishikawa cells. However, in the absence of SDF-1α stimulation （baseline）, the pAKT level in Ishikawa-PTEN cells was less than that in Ishikawa cells. There was a significant difference in growth between the Ishikawa-PTEN cells and the Ishikawa-neo cells. Conclusions PTEN gene transfection can regulate the level of pAKT but not pERK in Ishikawa-PTEN cells. PTEN protein may suppress the growth-promoting effect of SDF-1α on endometrial carcinoma by inhibiting the PI-3K/AKT signal transduction pathway.     

安罗替尼对肺腺癌细胞株A549放射敏感性的影响及机制

  目的  研究安罗替尼对肺腺癌细胞放疗增敏作用及机制。  方法  采用安罗替尼处理人肺腺癌细胞株A549，使用CCK8法测定安罗替尼对细胞增殖的影响；采用克隆形成实验测定安罗替尼联合放疗对细胞的生长抑制作用；使用流式细胞术测定细胞周期和细胞凋亡的变化；采用免疫荧光测定安罗替尼联合放疗对细胞内DNA损伤的影响；采用Western blot检测细胞内DNA损伤标志因子DNA-PKcs的表达变化。  结果  安罗替尼对人肺腺癌细胞A549增殖具有抑制作用，第2、3、4、5天，（P < 0.05）。体外培养克隆形成实验显示，安罗替尼联合放疗组的准予剂量（Dq）、平均致死剂量（D0）及4 Gy照射时的存活分数（SF）均明显低于单纯放疗组（P < 0.05）。安罗替尼能够使细胞阻滞于G1/G0期，与放疗联合作用后，进一步降低了G2和S期细胞比例。安罗替尼能够增加放疗诱导的细胞凋亡比例（31.94±2.25）%和（44.44±2.30）%，（P < 0.001），增加γH2AX阳性细胞数量（25.67±2.52）%和(54.67±3.79）%，（P < 0.001）。安罗替尼能够降低放疗诱导的DNA断裂损伤标志因子DNA-PKcs的表达强度（0.90±0.06）和（0.40±0.06），（P < 0.001）。  结论  安罗替尼具有放疗增敏作用，其机制可能与减少G2/S期细胞、增加细胞凋亡和维持DNA持续损伤有关。     

电离辐射对沉默ATRX的HeLa细胞DNA损伤修复的影响

目的：靶向沉默宫颈癌HeLa细胞中α-地中海贫血/精神发育迟滞综合征X染色相关蛋白（ATRX）,检测电离辐射对ATRX、γH2AX和Rad51蛋白表达及γH2AX和Rad51焦点数的影响,探讨ATRX参与辐射后HeLa细胞DNA损伤修复的作用。方法： 3条ATRX-shRNA和阴性对照（Control-shRNA）的慢病毒载体转染293T细胞,收集慢病毒并感染HeLa细胞,利用puromycin筛选获得稳定沉默ATRX的细胞系,分别命名为shA₁-HeLa、shA₂-HeLa、shA₃-HeLa和shCon-HeLa,采用Western blotting法检测沉默ATRX效率以及电离辐射后ATRX、γH2AX和Rad51蛋白的表达,采用免疫荧光技术观察shCon-HeLa和shA₁-HeLa组中γH2AX和Rad51焦点并计数其数量。结果： shCon-HeLa细胞中可见ATRX蛋白表达,而shA₁-HeLa、shA₂-HeLa和shA₃-HeLa细胞中均无ATRX蛋白表达,表明沉默效率较高。在2和8 Gy剂量照射后1、6和24 h,shCon-HeLa组ATRX蛋白表达量逐渐升高,24 h时表达量最高,且8 Gy照射后1、6和24 h表达量均较高。4 Gy照射后0~6 h,与shCon-HeLa组比较,shA₁-HeLa组γH2AX焦点数在1 h明显升高（P<0.05）,而后逐渐降低,但在6 h焦点数仍明显高于shCon-HeLa组（P<0.01）;Rad51焦点数与γH2AX焦点数变化相一致,与shCon-HeLa组比较,shA₁-HeLa组Rad51焦点数在1 h明显升高（P<0.05）,在6 h时shA₁-HeLa焦点数仍明显高于shCon-HeLa组（P<0.01）。4 Gy照射后0~16 h,shA₁-HeLa组细胞中γH2AX和Rad51蛋白表达量均较shCon-HeLa组增加。结论：成功获得稳定沉默ATRX的HeLa细胞模型,电离辐射可诱导ATRX蛋白表达量增加,且沉默ATRX的HeLa细胞中γH2AX和Rad51焦点数及蛋白表达量均高于对照组,提示ATRX参与了辐射诱导的DNA损伤修复过程。     

ABT737通过延迟宫颈癌细胞放射后DNA损伤修复及诱导凋亡而增敏放疗

【目的】 探讨类似BH-3的小分子物质ABT737诱导增敏放疗的作用及其分子机制?【方法】噻唑蓝法(MTT)检测不同浓度ABT737对HeLa细胞的生长抑制作用;细胞克隆形成实验检测ABT737联合放疗对放疗的增敏作用;免疫荧光法检测γ-H2AX观察DNA损伤修复情况;流式细胞术检测细胞凋亡;免疫印迹法检测Caspase-3?PARP的表达观察凋亡?【结果】与对照组相比,ABT737能显著抑制宫颈癌HeLa细胞的增殖(P < 0.05),并呈浓度依赖性,其IC50为15.7 μmol/L?体外培养克隆形成实验结果显示放疗+ABT737不同用药时间组的DEF值均大于1,放疗+8 μmol/L ABT737持续用药组为1.88,放疗+12 μmol/L ABT737用药72 h组为1.13,细胞存活分数(SF值)持续用药组为0.84,用药72 h组SF值为0.82;免疫荧光结果显示ABT737联合放疗处理HeLa细胞1 h后,放射线导致的γ-H2AX聚焦点数量及有γ-H2AX聚焦点生成的细胞数量均明显增加,上述处理24 h后,单纯放疗组γ-H2AX焦点消失,而联用ABT737处理组仍可观察到γ-H2AX焦点聚集?流式细胞术结果显示,单纯放疗组早期凋亡率(Annexin V+,PI-)为23.3%,ABT737联合放疗可以明显提高放射线诱导的细胞凋亡,早期凋亡率最高达50.3%?免疫印迹结果显示10 ?滋mol/L ABT737与2 Gy放射联合作用于Hela细胞后,凋亡蛋白cleaved Caspase-3与cleaved PARP的表达较单纯放疗组增加?【结论】 ABT737对宫颈癌Hela细胞具有放疗增敏作用,其机制与ABT737可延迟宫颈癌细胞放疗后DNA损伤修复及诱导凋亡有关?     

沉默MSH3表达增强舌癌细胞对顺铂的敏感性

目的设计靶向DNA错配修复基因MSH3的siRNA干扰片段,观察MSH3有效沉默对舌癌细胞顺铂耐药性的影响。方 法针对MSH3基因CDS区序列,设计3条siRNA片段,体外化学合成小干扰RNA（siRNA）片段,通过脂质体介导转染人舌癌细 胞CAL27。Real-time PCR及Western 检测干扰后MSH3 的表达。MTS,凋亡双染法及细胞免疫荧光检测顺铂处理后细胞存 活、凋亡及DNA双链断裂的情况。结果3 条siRNA 片段转染细胞后,与对照组相比,3 号siRNA 片段转染后能显著降低 MSH3 mRNA及蛋白质表达水平,该片段被选为后续实验。MSH3表达下调后,顺铂的半数抑制浓度（IC50值）分别由21.32降 至13.95 μmol/L（P<0.05）,凋亡指数分别由4.23±1.27升至11.32±1.82（P<0.05）,舌癌细胞中γ-H2AX焦簇（Foci）的数量显著增 加。结论MSH3表达下调可以明显增加舌癌细胞对顺铂的敏感性,减少DNA双链断裂修复是其主要机制之一。     

肿瘤坏死因子-α对人肺癌A549细胞的放疗增敏作用

目的探讨肿瘤坏死因子-α(TNF-α)在人肺癌A549细胞放射治疗中的增敏作用。方法单独γ射线辐照细胞或联合应用TNF-α处理细胞,通过MTT法检测人肺癌A549细胞存活生长曲线,采用流式细胞术分别检测细胞凋亡和细胞周期的变化,并利用Western blot法分析细胞中Caspase-3蛋白表达的变化。结果与单独γ射线作用相比,TNF-α和γ射线联合应用能明显抑制人肺癌A549细胞的生长,同时诱导和促进A549细胞的凋亡发生,调整细胞周期的变化,这一过程与细胞中Caspase-3蛋白表达的增加相关。结论 TNF-α可以增加人肺癌A549细胞的放疗敏感性,提高γ射线对人肺癌A549细胞的放射治疗效果。     

SET8 Inhibition Potentiates Radiotherapy by Suppressing DNA Damage Repair in Carcinomas

Objective SET8 is a member of the SET domain-containing family and the only known lysine methyltransferase(KMT) that monomethylates lysine 20 of histone H4(H4 K20 me1). SET8 has been implicated in many essential cellular processes, including cell cycle regulation, DNA replication, DNA damage response, and carcinogenesis. There is no conclusive evidence, however, regarding the effect of SET8 on radiotherapy. In the current study we determined the efficacy of SET8 inhibition on radiotherapy of tum...     

蛋白激酶CK2α对喉癌细胞凋亡和超微结构的影响

目的 探讨蛋白激酶CK2α对人喉癌细胞凋亡和超微结构的影响及其可能机制.方法 用脂质体转染法分别将蛋白激酶CK2α特异性siRNA表达质粒psiRNA-hH1neo-CK2α及非特异性siRNA表达质粒psiRNA-hH1neo-cont转染人喉癌Hep-2细胞.Western印迹法检测转染细胞蛋白激酶CK2α蛋白表达,膜联蛋白V-异硫氰酸荧光素(Annexin V-FITC)和碘化丙啶(PI)双染色法检测转染细胞凋亡率的变化,透射电镜观察转染细胞形态学变化,Western印迹法检测转染细胞bcl-2和Bax蛋白的表达.结果 转染psiRNA-hH1neo-CK2α质粒后,Hep-2细胞蛋白激酶CK2α蛋白表达明显下降(P＜0.01).和未转染细胞组和psiRNA-hH1neo-cont转染细胞组比较,psiRNA-hH1neo-CK2α转染组细胞出现典型的凋亡征象,如核固缩、染色质凝集靠近核膜和凋亡小体形成.psiRNA-hH1neo-CK2α转染细胞凋亡率明显高于未转染细胞组和psiRNA-hH1neo-cont转染细胞组(25.66%±0.83%比3.66%±0.43%、5.18%±0.22%,均P＜0.05);与其他2组比较,psiRNA-hH1neo-CK2α转染组细胞bcl-2蛋白表达较低(相对吸光度比值为0.20±0.09 vs 0.72±0.16、0.56±0.11,均P＜0.01),Bax蛋白表达较高(相对吸光度比值为0.81±0.17 vs 0.26±0.12、0.33±0.17,均P＜0.01),bcl-2/Bax较低(0.25±0.05 vs 2.76±0.21、1.70±0.22,均P＜0.01).结论 蛋白激酶CK2α与喉癌细胞凋亡密切相关,该作用可能与bcl-2/Bax降低有关,蛋白激酶CK2α可能是一个有潜力的喉癌治疗靶点.     

TGF －β1／Smad3对辐射所致鼻咽癌细胞凋亡及细胞周期分布的影响

目的：探讨TGF－β1／Smad3在辐射所致鼻咽癌细胞CNE－2凋亡中的作用及其机制。方法 CNE－2细胞分为6 Gy照射组、TGF－β1＋6 Gy照射组以及对照组,免疫荧光方法检测γH2AX焦点形成试验,流式细胞仪检测凋亡细胞及细胞周期分布,MTS法检测细胞生长生长活力,绘制生长曲线并计算生长增殖率。 Western blot 检测P－ATM、pCHK1、pCHK2、CDC25A、p－Smad3、Smad3、P15和CDK4蛋白表达水平。结果照射组及TGF－β1＋6 Gy照射组凋亡细胞数明显增多,细胞核体积缩小,可见浓染致密的γH2AX表达。 TGF－β1＋6 Gy 照射组的γH2AX表达相对于对照组与6 Gy照射组明显增加。6 Gy照射组和TGF－β1＋6 Gy照射组的G2／M期细胞比例均明显高于对照组（P＜0.05）。 TGF－β1＋6 Gy照射组与6 Gy照射组相比,S期明显增加（P＜0.05）。 TGF－β1＋6 Gy照射组和6 Gy照射组的凋亡及坏死细胞比对照组的明显增加,差异有统计学意义（ P＜0.01）。 TGF－β1＋6 Gy照射组的凋亡和坏死细胞比例明显比6 Gy照射组增加,差异有统计学意义（ t P＜0.05）。对照组、6 Gy照射组和TGF－β1＋6 Gy组的CNE－2细胞克隆形成率分别为0.60、0.03和0.01,与对照组相比,6 Gy照射组和TGF－β1＋6 Gy照射组的细胞克隆形成明显减少,差异有统计学意义（ P＜0.01）。与6 Gy照射组相比,TGF－β1＋6 Gy照射组的细胞形成率减少,差异有统计学意义（ P＜0.01）。 Western blot 结果显示TGF－β1处理组相对于单独6 Gy照射组,p－ATM、p－CHK1、p－CHK2、P15、p－Smad3、CDC25A及CDK4表达下调,Smad3表达无明显变化。结论 TGF－β1可增加照射所致鼻咽癌细胞的凋亡,可能与增加S期细胞比例以及抑制Smad3磷酸化同时促进ATM磷酸化有关。     

PRMT1通过促进RRM2表达抑制鼻咽癌细胞凋亡

目的 探讨PRMT1能否通过促进RRM2抑制鼻咽癌细胞凋亡。方法 免疫组化和Western blot共同检测鼻咽癌和癌旁组织中PRMT1及RRM2的相对表达量;以人正常鼻黏膜上皮细胞系（HNEpC）作为对照,Western blot实验检测不同鼻咽癌细胞系中PRMT1和RRM2蛋白的相对表达量;对CNE-2细胞中PRMT1分别进行过表达和下调,过表达实验设阴性对照组（oe NC：CNE-2细胞中转染pcDNA3.1质粒）、过表达PRMT1组（oe PRMT1：CNE-2细胞中转染pcDNA3.1 PRMT1质粒）、下调实验设阴性对照组（si-NC：CNE-2细胞中转染NC无关序列）及PRMT1小干涉RNA组（si-PRMT1：CNE-2细胞中转染PRMT1小干涉RNA）,对CNE-2细胞中RRM2分别进行过表达和下调,过表达实验设阴性对照组（oe NC：CNE-2细胞中转染pcDNA3.1质粒）、过表达RRM2组（oe RRM2：CNE-2细胞中转染pcDNA3.1 RRM2质粒）、下调实验设阴性对照组（si-NC：CNE-2细胞中转染NC无关序列）及RRM2小干涉RNA组（si-RRM2：CNE-2细胞中转染RRM2小干涉RNA）。Western blot验证PRMT1和RRM2的表达关系;AnnexinV-FITC/PI凋亡检测试剂盒检测细胞凋亡;活性氧试剂盒检测细胞活性氧。结果 与癌旁组织和HNEpC细胞相比,鼻咽癌组织和CNE-2细胞中的PRMT1和RRM2相对表达量均显著增加（P<0.05）;Western blot证明PRMT1可促进RRM2表达,过表达PRMT1或RRM2,CNE-2细胞活性氧产生量与凋亡率均降低（P<0.05）,下调PRMT1或RRM2表达,CNE-2细胞活性氧产生量与凋亡率凋亡率均增加（P<0.05）。Western blot显示,过表达PRMT1或RRM2表达,Cleaved caspase-3和Cleaved caspase-8蛋白表达减少（P<0.05）,下调PRMT1或RRM2表达,Cleaved caspase-3和Cleaved caspase-8蛋白表达增加（P<0.05）;当同时下调PRMT1和过表达RRM2表达,与单独下调PRMT1组比,CNE-2细胞活性氧产生量与凋亡率均显著降低（P<0.05）,Cleaved caspase-3和Cleaved caspase-8蛋白表达均减少（P均<0.05）。结论 PRMT1通过促进RRM2表达影响CNE-2细胞凋亡。