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蛋白质p53

蛋白质p53的相关文献在1995年到2018年内共计370篇,主要集中在肿瘤学、基础医学、内科学 等领域,其中期刊论文369篇、会议论文1篇、专利文献113305篇;相关期刊115种,包括中国病理生理杂志、中华病理学杂志、医学临床研究等; 相关会议1种,包括贵州省第三届科技创新论坛暨第二届青年科技论坛等;蛋白质p53的相关文献由1445位作者贡献,包括于燕妮、张鹏飞、朱明华等。

蛋白质p53—发文量

期刊论文>

论文:369 占比:0.32%

会议论文>

论文:1 占比:0.00%

专利文献>

论文:113305 占比:99.67%

总计:113675篇

蛋白质p53—发文趋势图

蛋白质p53

-研究学者

  • 于燕妮
  • 张鹏飞
  • 朱明华
  • 李一伟
  • 陈一峰
  • 丁钢
  • 倪灿荣
  • 刘斌
  • 刘鹏
  • 卢虹
  • 期刊论文
  • 会议论文
  • 专利文献

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    • 田超; 王玥; 李吉平; 王声远; 夏冰; 杨越; 李昶; 王琪; 吴永会
    • 摘要: 目的 研究抗坏血酸(Ascorbic acid,AA)对镍染毒小鼠胚胎成纤维细胞(NIH/3T3)P53、Bcl-2蛋白表达的影响.方法 研究采用染毒浓度为50 μg/mL镍精炼尘诱导建立损伤模型,通过测定细胞相对存活率、超氧化物歧化酶(SOD)、乳酸脱氢酶(LDH)和谷胱甘肽过氧化物酶(GSH-Px)活力、过氧化氢(H2O2)和丙二醛(MDA)含量以及p53(野生型)、Bcl-2蛋白表达情况,探讨不同剂量抗坏血酸(25、50、100 mmol/L)对镍精炼尘诱导NIH/3T3细胞损伤的保护作用.结果 抗坏血酸Ⅲ组可以使NIH/3T3细胞存活率显著提高;细胞凋亡率降低;SOD和GSH-Px的活力明显增加,差异有统计学意义(P<0.05);同时可以使MDA和H2O2水平,以及细胞外LDH酶活力明显降低,差异有统计学意义(P<0.05).抗坏血酸Ⅱ、Ⅲ组Bcl-2蛋白表达水平均高于镍精炼尘组,差异有统计学意义(P<0.05);抗坏血酸各剂量组p53蛋白表达水平均低于镍精炼尘组,差异有统计学意义(P<0.05).结论 随着抗坏血酸浓度的增加,氧化损伤水平降低,抗氧化酶水平上升,细胞凋亡减少,p53表达降低,Bcl-2表达增加,表明抗坏血酸对镍精炼尘染毒NIH/3T3细胞损伤具有一定的保护作用.%Objective To study the protective effect of Ascorbic acid (AA) on the injury of nickelexposed mouse embryonic fibroblasts (NIH/3T3).Methods A model of damage induced by 50 μg/mL nickel refining dust was established to determine the relative survival rate of cells,superoxide dismutase (SOD),lactate dehydrogenase (LDH) and glutathione peroxidase.(GSH-Px) activity,hydrogen peroxide (H2O2) and malondialdehyde (MDA) content,and p53 (wild-type),Bcl-2 protein expression.To investigate the protective effect of different doses of ascorbic acid (25,50,100 mmol/L) on nickel-refined dust-induced NIH/3T3 cell injury.Results The study showed that ascorbic acid Ⅲ group can make the NIH/3T3 cell survival rate increased significantly;Apoptosis rate was reduced;The vitality of SOD and GSH-Px increased significantly,and the difference was statistically significant (P<0.05).At the same time,the level of MDA and H2O2 and the activity of extracellular LDH enzyme were significantly reduced,and the difference was statistically significant (P<0.05).The results showed that nickel refining dust induced cell damage through up-regulation of p53 protein and down-regulation of Bcl-2 protein expression;ascorbic acid interventions,the expression level of Bcl-2 protein in ascorbic acid Ⅱ and Ⅲ groups was higher than that of nickel refining dust group,and the difference was statistically significant (P<0.05);The expression level of p53 protein in each dose group of ascorbic acid was lower than that of nickel refined dust group,and the difference was statistically significant (P<0.05).Conclusion With the increase of concentration of ascorbic acid,oxidative damage levels,antioxidant enzyme levels,reduce cell apoptosis,reduce expression of p53,increased expression of Bcl-2.It showed that ascorbic acid had protective effect on NIH/3T3 cell injury induced by nickel refining dust.
    • 张云鹤; 李广太; 王红霞
    • 摘要: 目的 探讨外阴上皮内非瘤变(nonneoplastic epithelialdisorders of vulva,NNEDV)患者PTEN、p53蛋白质表达水平变化及意义,以了解其发病机制,寻求有效治疗方案.方法 选择外阴上皮内非瘤变组织40例,其中硬化性苔癣(LS)14例,鳞状上皮细胞增生(SH)21例,SH合并LS5例,另选择因外阴整形或阴道前后壁修补术者标本40例为正常对照组.应用免疫组化方法检测PTEN、p53蛋白质的表达,并分析其与外阴上皮内非瘤变发病的关系.结果 发现在正常外阴皮肤组织未能见p53蛋白表达,在NNEDV中表达率为25%,和正常皮肤比较,差异有显著性(P<0.05).LS和SH组织中,p53蛋白阳性表达率分别为35.7%和23.8%,两者比较,差异无显著性(P>0.05);PTEN在正常皮肤中的表达100%,而在NNEDV组织中仅仅85% (P=0.026),显示差异有显著性.LS和SH组织中PTEN蛋白阳性表达率分别为76.2%和92.8%,两者比较,差异无显著性(P>0.05).结论 NNEDV组织中PTEN和p53蛋白异常表达,推测PTEN和p53基因参与了外阴异常增殖过程,其突变或缺失在NNEDV甚至外阴鳞癌的发生、发展过程中起重要作用.
    • 万玲玲; 梁芸; 李玉雪; 梁翠娟; 李立新
    • 摘要: Objective To investigate the clinical diagnosis value of P16 gene methylation and P53 antibody in serum for non-small cell lung cancer (NSCLC).Methods The P16 gene methylation and P53 antibody level in serum from 98 NSCLC patients and 60 normal controls were tested with methylationspecific polymerase chain reaction and enzyme linked immunosorbent assay.Results The sensitivities of P16 gene methylation and P53 antibody qualitative detection were 70.41% and 58.16% for prediction diagnosis of NSCLC,the specificities were 85.00% and 93.33%,the correct indexes were 0.554 and 0.515.The positive rates of P16 gene methylation in serum of NSCLC patients and healthy controls were 70.41% and 15.00% (x2 =45.709,P <0.001).The positive rates of P53 antibody in serum of NSCLC patients and healthy controls were 58.16% and 6.67% (x2=41.638,P <0.001).The result of receiver operating characteristic curve in the diagnosis of NSCLC showed that the quantitative detection of serum P53 antibody in the diagnosis of NSCLC was statistically significant.The sensitivity and specificity of combined detection of P16 methylation and P53 antibody were 82.56% and 78.33% for NSCLC diagnosis,the correct index was 0.610.The sensitivity of combined detection of P16 methylation and P53 antibody in NSCLC was higher than that of single detection (x2=14.105,P =0.001).Conclusions The detection of P16 gene methylation and P53 antibody in serum of NSCLC patients is of practical significance.The sensitivity of combined detection is higher than that of single detection.%目的 探讨血清P16基因甲基化与P53抗体水平的检测对非小细胞肺癌(NSCLC)的诊断价值和临床意义.方法 选择98例NSCLC患者为实验组,以60名健康者作为对照,分别采用甲基化特异性PCR法检测其血清P16基因甲基化,用酶联免疫吸附试验法检测其血清P53抗体.结果 P16基因甲基化和P53抗体定性检测对NSCLC预测诊断的敏感度分别为70.41%和58.16%,特异度分别为85.00%和93.33%,正确指数为0.554和0.515;P16基因甲基化在NSCLC患者和健康对照者血清中阳性率分别为70.41%和15.00% (x2=45.709,P<0.001);P53蛋白抗体在NSCLC患者和健康对照者血清中阳性率分别为58.16%和6.67%(x2=41.638,P<0.001);对NSCLC诊断做ROC曲线结果显示血清学P53抗体定量检测对NSCLC预测诊断有统计学意义;P16基因甲基化和P53抗体定性联合检测结果显示对NSCLC诊断的敏感度为82.56%,特异度为78.33%,正确指数为0.610.P16基因甲基化和P53蛋白抗体联合检测的敏感度高于单独检测(x2=14.105,P=0.001).结论 血清中P16基因甲基化与P53抗体的检测对NSCLC患者的预测诊断具有实际意义,联合检测的敏感度高于单独检测,可在临床上予以推广.
    • 杨宣琴; 步鹏; 连婧; 解立武; 徐恩伟
    • 摘要: Objective To investigate the expression of metadherin (MTDH) and p53 gene in esophageal squa-mous cell carcinoma (ESCC) and its relationship with clinicopathological features. Methods Immunohistochemistry was used to determine the expression levels of MTDH and P53 protein in the tissues of 78 cases of ESCC, and 25 cases of intraepithelial neoplasia and normal esophageal mucosa. Results The positive expression rate of MTDH in the tissues of normal esophageal mucosa, high grade intraepithelial neoplasia and ESCC was 8%, 64% and 72%, re-spectively (P0.05). The protein expression of MTDH and P53 were not related to age, gender and invasion depth of patients. Conclusion The overexpression of MTDH and P53 protein is closely related to the occurrence and development of ESCC, which play an important role in the invasion and metastasis of esophageal cancer, and can be used as an important indicator for the prognosis of esophageal cancer.%目的:研究异黏蛋白(MTDH)和p53基因在食管鳞状细胞癌中的表达及其与临床病理特点之间的关系。方法采用免疫组织化学方法检测78例食管鳞癌患者的食管鳞状细胞癌组织、25例上皮内瘤变和25例正常食管黏膜组织中MTDH和P53蛋白的表达情况。结果 MTDH在正常食管黏膜组织、高级别上皮内瘤变、食管鳞状细胞癌组织中的阳性表达率分别为8%、64%和72%(P<0.01),MTDH与食管鳞状细胞癌的分化程度、淋巴结转移差异有统计学意义(P<0.05)。P53蛋白在正常食管黏膜组织、上皮内瘤变、食管鳞状细胞癌组织中的表达亦逐渐升高,分别为4%、48%、58%(P<0.01)。 P53蛋白与食管鳞状细胞癌组织分化程度差异有统计学意义(P<0.05),有淋巴结转移食管癌患者P53蛋白阳性率高于无淋巴结转移者(65%、47%),但2组间比较差异无统计学意义(P>0.05)。 MTDH和P53蛋白的表达与患者年龄、性别、浸润深度无关。结论 MTDH和P53蛋白的过度表达与食管鳞状细胞癌的发生、发展密切相关,在食管癌的侵袭和转移过程中发挥了重要作用,可作为预测食管癌预后的重要指标。
    • 甘伟平; 罗陈飘; 陈喜志; 吴东海; 王立生
    • 摘要: 目的 分析研究P21、P27和P53蛋白在结肠癌组织中的表达水平及关联.方法 选择2011年2月至2014年11月来我院就诊的144例经外科手术切除的原发性结肠癌组织标本进行分析研究.采用免疫组织化学染色方法进行染色,根据相关评价标准计算出阳性细胞率和染色强度,并最终确定阳性率等级.结果 P21蛋白在有淋巴结转移的结肠癌组织中的阳性率达70.5%,高于无淋巴结转移的结肠癌的阳性率(30.3%),差异具有统计学意义(P<0.05),与其他临床病理因素的关联性很小(P>0.05).P27蛋白在无淋巴结转移的结肠癌组织中的阳性率达84.8%,高于有淋巴结转移的结肠癌的阳性率(55.1%),差异具有统计学意义(P<0.05);在结肠癌高中分化阳性率(80.6%)高于低分化阳性率(69.4%),差异具有统计学意义(P<0.05),但是与其他临床病理因素的关联性很小(P>0.05).P53蛋白在结肠癌组织的阳性表达率与TNM分期、分化程度及浸润深度相关性大(P<0.05),但是与其他临床病理因素的关联性小(P>0.05).结论 P21、P27、P53蛋白表达可以作为结肠癌的辅助诊断指标,3种蛋白在判断结肠癌的生物学行为上具有协同作用.
    • 原向伟; 黄秀芳; 陈忠羡; 梁胜根; 廖威明
    • 摘要: Objective To investigate the effect of IFNαon doxorubicin-induced apoptosis in human osteosarcoma cells with its molecular mechanisms to provide evidences for improving the treatment of osteosarcoma.Methods Osteosacoma U2OS and MG63 cells were treated with IFNαand Doxorubicin, alone or in combination, for 72 h .Cytotoxicity was determined with MTT.Apoptosis was evaluated through fluorescence-activated cell sorting, Hoechst33258 staining and DNA ladder assay.The expression of p53, Bax, Bcl-2, Mdm2, p21, caspase-3 and PARP was determined with Western blot.siRNA interference was used to silence p53.Results IFNαtreatment for 72 h did not induce cytotoxicity but greatly enhanced doxorubicin-induced cytotoxicity and apoptosis in p53-wild U2OS cells but not p53-mutant MG63 cells. Compared with other groups, the combination of IFNαand doxorubicin induced more obvious apoptotic morphological changes and DNA ladder.IFNαdid not alter the expression of the indicate genes.The expression of p53, Bax, Mdm2 and p21 was up-regulated by doxorubicin and further increased in response to combination.The expression of Bcl-2 was down-regulated by doxorubicin and further decreased in response to combination.There were no differences among groups in MG63 cells.The expression of p53 was effectively blocked by p53-siRNA in U2OS cells.The p53 silencing greatly reduced the cytotoxicity mediated by combination for 72 h, compared with non-and control-siRNA groups.The activation of caspase-3 and PARP mediated by combination was largely suppressed by p53 silence.Conclusion IFNαsensitizes human osteosarcoma cells to doxorubicin-induced apoptosis through p53-dependent pathway.The combination of IFNαand traditional chemotherapy can be used in osteosarcoma treatment.%目的:探讨α干扰素(IFNα)对阿霉素诱导骨肉瘤细胞凋亡的影响及其分子机制。方法采用α干扰素(IFN)和阿霉素单独或联合处理骨肉瘤 U2OS 和 MG63细胞72 h,通过 MTT 法测定细胞生长抑制,应用流式细胞术、Hochest 33258荧光染色和 DNA Ladder 检测细胞凋亡,免疫印迹法检测聚腺苷酸二磷酸核糖转移酶(PARP)、凋亡相关蛋白(p53、Bax、Bcl-2、Mdm2、p21、Caspase-3)和PARP 的表达,实时聚合酶链反应(RT-PCR)检测 p53在 mRNA 水平的表达,通过 siRNA 干扰进行 p53的表达沉默。结果IFNα本身对 U2OS 和 MG63无细胞毒作用,但在 p53正常的 U2OS 而非 p53突变的 MG63细胞中增强阿霉素引起的细胞毒作用和凋亡,在 IFNα+阿霉素组的 U2OS 细胞出现更明显的凋亡形态学改变和 DNA 梯形条带。在 U2OS 细胞中,IFNα对各基因的表达无影响,p53、Bax、Mdm2、p21可被阿霉素上调,并被联合用药进一步增强。而 Bcl-2在阿霉素作用后下调,并被联合用药进一步下调,在 MG63细胞中,各基因的表达无明显变化。在 U2OS 细胞中转染 p53-siRNA 阻断了p53的表达,经联合用药处理72 h 后,较空白组和对照 siRNA 组,转染 p53-siRNA 组的细胞活力明显增高明显,并且相对于对照 siRNA 组,联合用药引起的 caspase-3和 PARP 的活化也在 p53-siRNA 组明显减弱。结论IFNα可通过 p53依赖性凋亡途径增强阿霉素诱导的骨肉瘤细胞凋亡,联合 IFNα生物治疗和化疗可望应用于骨肉瘤的治疗。
    • 张永侠; 张彬; 周慧芳; 刘文胜; 陈汶; 徐震纲
    • 摘要: Objective To analyze the clinical characteristics,prognosis and molecular biological changes of tonsillar squamous cell carcinoma(TSCC).Methods Retrospective analysis of 61 TSCC cases treated from January 1999 to December 2012.Demographic data and clinical charts,including histologic grade of tumor,treatment and outcome of the patients,were reviewed.Human papillomavirus (HPV)-DNA were detected using SPF10-DNA enzyme immunoassay and LiPA genotyping method.Expressions of p16 and p53 proteins were examinated by immunohistochemistry.Survival rate was calculated with SPSS 19.0 software using the Kaplan-Meier method.Results There were 55 males and 6 females,with a median age of 57 years.Of the 61 TSCC,21 were with well differentiation,19 with moderate differentiation and 21 with poor differentiation,including 7 patients at stage Ⅱ,10 at stage Ⅲ and 44 at stage Ⅳ.HPV-positive rate of TSCC was 29.5% (18/61) and high-risk HPV-16 subtype accounted for 72.2% (13/18).The percentage of famel patients in HPV-positive TSCC was higher than HPV-negative TSCC(22.2% vs 4.7%).HPV-positive TSCC was more common in non-smoking patients (50.0% vs 79.1%,x2 =5,155,P =0.023) and non-drinking patients (27.8% vs 51.2%,x2 =4.346,P =0.037).HPV-positive TSCC mostly presented with high expression of p16 protein (88.9% vs 16.3%,x2 =28.481,P =0.000),and low expression of p53 protein(72.7% vs 46.5%,x2 =5.028,P =0.025).The prognosis of patients with HPV-associated TSCC was significantly better than non-HPV-associated TSCC,and The 3-year and 5-year overall survival rates of patients with HPV-positive TSCC were higher than those of patients with HPV-negative TSCC (87.7% vs 49.5% and 78.9% vs 33.0%,respectively).Conclusion HPV-associated TSCC had unique clinicopathological and molecular biological features,showing better prognosis compared to HPV-negative TSCC.%目的 总结分析人乳头状瘤病毒(human papillomavirus,HPV)相关扁桃体鳞状细胞癌(鳞癌)患者的临床病理特征、预后特点及分子生物学改变.方法 回顾性分析1999年1月至2012年12月期间中国医学科学院肿瘤医院收治的符合入组条件的61例扁桃体鳞癌患者的临床病理资料,应用HPV SPF10 PCR/DEIA检测和LiPA分型方法进行HPV-DNA检测及分型;采用免疫组织化学法检测p16及p53蛋白表达情况.采用Kaplan-Meier法计算生存率.结果 61例扁桃体鳞癌患者中,男55例,女6例;中位发病年龄57岁.肿瘤临床分期为Ⅰ期0例,Ⅱ期7例,Ⅲ期10例,Ⅳ期44例.肿瘤的分化程度为高分化21例,中分化19例,低分化21例.扁桃体鳞癌HPV总感染率为29.5%(18/61),男性14例,女性4例,其中HPV-16亚型占全部感染的72.2% (13/18).HPV相关扁桃体鳞癌女性患者比例高于非HPV相关扁桃体鳞癌(22.2%比4.7%),且更易发生于非吸烟及非饮酒患者(吸烟:50.0%比79.1%,x2=5.155,P=0.023;酗酒:27.8%比51.2%,x2=4.346,P=0.037).HPV相关扁桃体鳞癌p16蛋白多为高表达(88.9%比16.3%,x2=28.481,P=0.000),而p53蛋白多为低表达(72.7%比46.5%,x2=5.028,P=0.025).Kaplan-Meier法计算HPV相关扁桃体鳞癌3年和5年总生存率分别为87.7%和78.9%,明显高于非HPV相关扁桃体鳞癌的3年总生存率49.5%和5年生存率33.0%,差异有统计学意义(x2=9.139,P=0.003).结论 HPV相关扁桃体鳞癌具有特殊的临床病理特征及分子生物学改变,且预后显著好于非HPV相关扁桃体鳞癌.
    • 魏炜; 肖康; 张宝云; 张乃嵩; 董小平; 石琦
    • 摘要: Objective To analyze the expression,mutation and codon 72 polymorphism in the Chinese laryngeal squamous cell carcinoma patients and the relationship with HPV infection.Methods The paraffin-embedded tissue blocks from 64 pathological-diagnosed laryngeal squamous cell carcinoma patients were included in this study.PCR and direct sequencing were used to screen the mutations in TP53 gene and immunohistochemistry was used to evaluate the expressing profiles of p53 proteins.Functional analysis software online were used to analyze the function changes of mutated p53 proteins.The relationship between TP53 mutation and phenotypes of SNP72 with other features of the patients was analyzed by Chisquare and Fisher' s exact test.Results Out of 64 laryngeal squamous cell carcinoma patients,sequencing confirmed 30 TP53 mutations in 22 cases (73.3%) and p53 immunohistochemistry (IHC) confirmed 20 (31.3%) positive staining.Online software analysis showed that most of the mutated p53 proteins lost their normal function.Analyses of SNP72 polymorphism revealed that the patients with Arg phenotype had significantly older onset ages than those with Pro phenotype but without statistic difference.Conclusion TP53 mutations are frequently observed in Chinese patients with laryngeal squamous cell carcinoma and this is one of the pathogenesis for laryngeal squamous cell carcinoma.%目的 了解喉鳞状细胞癌患者p53表达、突变,72位氨基酸多态性与HPV感染的关系方法 在该研究中收集了64份石蜡包埋的病理标本,用PCR的方法对p53基因进行扩增、测序和分析,用免疫组织化学的方法对p53蛋白进行检测,并且应用功能分析软件对突变蛋白进行功能分析.应用统计学软件分析p53的突变与72位密码子多态性以及其他因素的关系.结果 在64例临床诊断的喉癌病例中,通过PCR及测序的方法发现在p53基因高变区内22例病例具有30个突变.利用免疫组织化学的方法发现20例病例出现阳性改变.通过功能预测发现,大部分突变后的p53蛋白均丧失了其功能,只有4个突变点没有影响功能.72位密码子为精氨酸组的病例年龄(60岁)比脯氨酸组的病例年龄(55岁)偏大,但无统计学差异.结论 喉癌病例中p53基因的突变是常见的现象,是喉癌发生的原因之一.
    • 张馨; 毛大华; 邓英蕾
    • 摘要: 目的:探讨人乳腺癌组织中多药耐药相关蛋白(MRP)与P53蛋白的表达及关系.方法:采用免疫组化方法检测43例手术切除的乳腺癌标本中MRP与P53蛋白的表达,17例乳腺纤维腺瘤标本作为对照.结果:乳腺癌组织中MRP、P53蛋白的阳性率显著高于乳腺纤维腺瘤组(P<0.05);MRP的表达水平与年龄、月经状况、肿瘤大小、有无淋巴结转移、病理类型及TNM分期无关(P>0.05);P53蛋白的表达在有淋巴结转移时显著高于无淋巴结转移时,与年龄、月经状况、肿瘤大小、病理类型及TNM分期无关(P>0.05);MRP的阳性表达率与P53蛋白表达高低呈正相关(r=0.396).结论:MRP表达程度不受乳腺癌病程发展的影响,P53蛋白与乳腺癌的发生、发展有关,对评估患者预后可能具有重要意义,P53蛋白对乳腺癌中MRP的表达可能起调控作用.
    • 原向伟; 黄秀芳; 陈忠羡; 梁胜根; 廖威明
    • 摘要: Objective To explore the effects of tumor suppressor p14ARF on chemosensitivity of human osteosarcoma U2OS cells to cisplatin and elucidate its molecular mechanism.Methods U2OS cells expressing no p14ARF and U2OS-ARF cells expressing p14ARF stably through stable transfection were treated with cisplatin.Cell viability and IC50 were assayed with methyl thiazolyl tetrazolium (MTT).Apoptosis was examined by fluorescence-activated cell sorting and Hoechst33258 staining.The expressions of p53,Bax,p21,Mdm2 and Fas were detected by Western blot.And colorimetry was used to determine the activities of caspase-3,caspase-8 and caspase-9.Results The viability was 84.8% ± 4.4%,86.9% ± 5.0% and 66.7% ±4.6% respectively in U2OS,U2OS-vec and U2OS-ARF cells.The values of IC50 were (15.8 ± 0.9) μmol/L,(16.3 ± O.6) and (8.9 ± 0.8) μmol/L respectively in U2OS,U2OS-vec and U2OS-ARF cells.The levels of viability and IC50 obviously decreased in U2OS-ARF cells in response to cisplatin (P < 0.05).There were higher apoptotic rate and more obvious apoptotic morphological changes in U2OS-ARF cells than U2OS and U2OS-vec cells.The basal levels of p53,Mdm2 and p21 in U2OS-ARF cells were slightly higher than those in U2OS-vec cells.Cisplatin up-regulated p53,Mdm2 and p21 in both cell lines.However,the up-regulation was more pronounced in U2OS-ARF cells.Cisplatin did not change the levels of Bax and Fas in U2OS-vec cells.Bax protein was up-regulated in U2OS-ARF cells while the level of Fas remained constant,p14ARF also enhanced the activities of caspase-9 and caspase-3 in response to cisplatin.Conclusion p14ARF enhances the chemosensitivity to cisplatin in human osteosarcoma U2OS cells through p53 apoptotic pathway.And intrinsic mitochondrial apoptosis is involved.%目的 探讨抑癌基因p14ARF增强骨肉瘤U2OS细胞对顺铂的敏感性及其作用机制.方法 使用顺铂处理不表达p14ARF的U2OS细胞和稳定表达p14ARF的U2OS-ARF细胞,采用噻唑蓝(MTT)比色法测定细胞毒作用和半数抑制浓度值;流式细胞术和Hoechst 33258荧光染色检测细胞凋亡;免疫印迹法检测p53以及其下游基因Bax、p21、Mdm2、Fas的表达;比色法检测半胱氨酸天门冬氨酸蛋白酶(caspase)-3、caspase-8、caspase-9蛋白酶的活性.结果 顺铂处理72 h后,U2OS、U2OS-vec和U2OS-ARF组的细胞活力分别为(84.8%±4.4%)、(86.9%±5.0%)和(66.7%±4.6%),相比U2OS-ARF的细胞活力明显降低,U2OS、U2OS-vec和U2OS-ARF组的IC50分别为(15.8±0.9)、(16.3±0.6)μmol/L和(8.9±0.8)μmol/L,相比U2OS-ARF的IC50明显降低(P<0.05).流式细胞术和形态学鉴定表明相对于U2OS和U2OS-vec细胞,U2OS-ARF细胞表现更为明显的凋亡比例和凋亡特异性形态学变化.在U2OS-ARF细胞中,p53、Mdm2和p21的基础表达水平稍稍高于U2OS-vec对照细胞,顺铂处理在U2OS-vec和U2OS-ARF细胞均明显激活p53、Mdm2和p21的表达,但在U2OS-ARF细胞中更加明显;另外,顺铂处理对U2OS-vec细胞中Bax和Fas的表达没有影响,却在U2OS-ARF细胞中明显增强了Bax的表达,但对Fas无影响.p14ARF还增强顺铂对caspase-9和caspase-3的活化.结论 p14ARF通过p53凋亡通路增强骨肉瘤U2OS细胞对顺铂的敏感性,并与内源性线粒体通路有关.
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