HPV16 E6/E7 Negatively Affect Radiosensitivity of Lung Cancer Cells
Lu Lu, Qinghui Meng, Ming Cui, Xiaofei Chu, Shuyi Zhao, Huiwen Xiao, Jiali Dong, Eliot Marty Rosen, Saijun Fan
2016, 40(2): 85-90. doi: 10.3760/cma.j.issn.1673-4114.2016.02.001
Objective Lung cancer cells associated with radioresistance are likely to give rise to local recurrence and distant metastatic relapse, but little is known about its underlying mechanisms. In the present paper, the effects of the HPV16 E6 and HPV16 E7 oncoprotein on the radiosensitivity of lung cancer cell lines were investigated. Methods The HPV16 E6 or HPV16 E7 oncoprotein was expressed by a transient transfection with pcDNA3-HPV16 E6 or pcDNA3-HPV16 E7 expression vector. Human lung cancer H2179 cells and mouse lung cancer Lewis cells were exposed to a γ-ray radiation source, cellular survival was evaluated by using a colony formation assay. The expression of HPV16 oncoproteins E6/E7, extracellular signal-regulated kinases 1/2(ERK1/2) and AKT signaling was determined by Western blot assay. VEGF secretion was determined by ELISA. Results Both HPV16 oncoproteins E6 and E7 significantly decreased radiosensitivity of H2179 cells, associated with a promotion of the ERK1/2 and AKT phosphorylation. A decrease of reactive oxygen species(ROS) and an increase of VEGF levels were observed in the cells expressing the HPV16 oncoproteins E6 and E7. Furthermore, a similar reduction of radiosensitivity mediated by the HPV16 oncoproteins E6 and E7 was also observed in a mouse lung cancer Lewis cells. Conclusion The findings indicate that the HPV16 oncoproteins E6 and E7 negatively affects susceptibility of lung cancer cells to radiotherapy via regulation of the ERK1/2 and Akt signaling pathway and VEGF expression.
关键词: Papillomavirus, human, Lung neoplasms, Radiosensitivity, ERK1/2 and AKT, ROS
RBAP96 Mediates Radiosensitivity of Breast Cancer Cells via Interacting with Retinoblastoma Protein
Junling Zhang, Xiaolei Xue, Qinghui Meng, Lu Lu, Ming Cui, Saijun Fan
2016, 40(5): 323-328. doi: 10.3760/cma.j.issn.1673-4114.2016.05.001
Objective To identify a novel retinoblastoma protein(RB)-associated protein(RBAP 96) and to explore the impact of RBAP96 on radiosensitivity of human breast cancer cells. Methods An in vivo and in vitro association of RBAP96 with RB was determined by immunoprecipitation-Western blotting and GST pull-down assay. Protein expression was measured by Western blot assay. Cellular survival was evaluated by using a colony formation assay. Results In both in vitro and in vivo assays, we found that the RBAP96 and RB interaction required a 513LXCXE517 motif on the RBAP96 protein and an intact A/B binding pocket of RB. RBAP96 enhances RB-mediated transcriptional repression. Finally, enforced expression of RBAP96 caused an elevated radiosensitivity of human breast cancer cells bearing wtRB, but did not affect radiosensitivity of breast cancer cells bearing mutant RB. Expression of a full-length RBAP96 with an 513LXCXE517 inactivating mutation(LXCXE→RXRXH) failed to result in any radiosensitivity alteration. Conclusion This study for the first time characterizes a novel RB-interacting protein RBAP96 and demonstrates that enforced expression of RBAP96 causes an increase of RBAP96-mediated transcription activation and radiosensitivity via a RB-interacting dependent manner.
关键词: RBAP96, Retinoblasloma protein, Breast neoplasms, Transcription repression, Radiation tolerance
Ionizing Radiation Induces HMGB1 Cyto-plasmic Translocation and Extracellular Release
Lili Wang, Li He, Guoqiang Bao, Xin He, Saijun Fan, Haichao Wang
2016, 40(2): 91-99. doi: 10.3760/cma.j.issn.1673-4114.2016.02.002
Objective A nucleosomal protein, HMGB1, can be secreted by activated immune cells or passively released by dying cells, thereby amplifying rigorous inflammatory responses. In this study we aimed to test the possibility that radiation similarly induces cytoplasmic HMGB1 translocation and release. Methods Human skin fibroblast (GM0639) and bronchial epithelial (16HBE) cells and rats were exposed to X-ray radiation, and HMGB1 translocation and release were then assessed by immunocytochemistry and immunoassay, respectively. Results At a wide dose range(4.0-12.0 Gy), X-ray radiation induced a dramatic cytoplasmic HMGB1 translocation, and triggered a time- and dose-dependent HMGB1 release both in vitro and in vivo. The radiation-mediated HMGB1 release was also associated with noticeable chromosomal DNA damage and loss of cell viability. Conclusions Radiation induces HMGB1 cytoplasmic translocation and extracellular release through active secretion and passive leakage processes.
关键词: X-ray, HMGB1, Tumor cells, Inflammatory response, Damage-associated molecule pattern molecules
降低B7-H3蛋白对受照肺癌细胞A549细胞周期和凋亡的影响
董佳丽, 罗丹, 李源, 肖惠文, 路璐, 崔明, 樊赛军
2017, 41(3): 193-198. doi: 10.3760/cma.j.issn.1673-4114.2017.03.007
目的观察用小干扰RNA(siRNA)敲降B7-H3蛋白的表达对人肺癌细胞A549的细胞周期和凋亡的影响。 方法培养人肺癌A549细胞,将B7-H3蛋白siRNA瞬时转染于A549细胞(称为siB7-H3转染组)。实验分为4组,即对照组、siB7-H3转染组、照射组、照射+siB7-H3转染组。采用137Cs γ射线一次性照射,照射剂量为4 Gy;用Western blotting法和实时定量PCR分别检测B7-H3蛋白和mRNA的表达;流式细胞仪检测细胞周期和细胞凋亡的改变。 结果与对照组相比,siB7-H3转染组的B7-H3蛋白水平明显降低,mRNA表达量也明显低于对照组且差异有统计学意义(t=-4.222,P=0.013)。siB7-H3转染组细胞的G0/G1期细胞阻滞,S和G2/M期细胞数量减少;与对照组相比,照射组出现轻微的G0/G1期阻滞和明显的G2/M期细胞阻滞,照射+siB7-H3转染组的G0/G1、G2/M期均有明显的阻滞。照射后48 h,与对照组相比,照射组细胞的坏死率和凋亡率明显升高,siB7-H3转染组和照射+siB7-H3转染组的坏死率和凋亡率均无明显改变。 结论降低肺癌A549细胞中B7-H3蛋白表达水平可明显增加辐射诱导的G0/G1期阻滞,从而提示B7-H3表达水平的改变可能通过调节G0/G1细胞周期检查点而对肺癌细胞放射敏感性产生重要的调节功能。
关键词: 肺肿瘤, 癌, 非小细胞肺, 细胞周期, 细胞凋亡, B7-H3, siRNA
应用生物信息学确定结直肠癌辐射抗性细胞的差异表达基因
朱长春, 冯国兴, 董佳丽, 姜勉, 贺俊博, 樊赛军
2018, 42(4): 340-345, 351. doi: 10.3760/cma.j.issn.1673-4114.2018.04.010
目的基于生物信息学的方法,筛选结直肠癌辐射抗性细胞中的差异表达基因,从分子水平上初步探讨与结直肠癌抗辐射相关的潜在基因。方法从基因芯片公共数据库(GEO)中下载耐辐射的结直肠癌细胞基因表达谱数据(GSE43206),并利用R语言中的limma包进行差异基因筛选。对差异基因中的编码基因分别进行基因本体论(GO)富集分析、京都基因和基因组百科全书(KEGG)通路分析以及蛋白相互作用(PPI)分析,进一步筛选出PPI网络中的关键基因。通过实时荧光定量PCR实验确定5 Gy γ射线照射后人结肠癌HCT116细胞中关键基因的mRNA相对表达水平。采用Student t-test检验进行统计学分析,P < 0.05表示差异有统计学意义。结果共筛选出101个差异基因,包含67个上调基因,34个下调基因。GO富集分析发现这些差异基因在细胞迁移、DNA复制等生物学过程中富集。KEGG通路分析证实这些差异基因主要富集在乏氧诱导因子1信号通路。通过构建PPI网络,筛选出NDRG1PAG1LRP1PIM1LDLRPLAUR共6个与结直肠癌抗辐射相关的潜在基因。实时荧光定量PCR实验结果显示,与照射前比较,照射后人结肠癌HCT116细胞中NDRG1PAG1LRP1PIM1LDLRNDRG1PAG1LRP1PIM1LDLR关键基因的mRNA表达量显著上升,差异均有统计学意义(t=49.981,P < 0.01;t=26.420、28.698、21.358、23.545,均P < 0.05;t=50.601,P < 0.01)。结论利用生物信息学能够快速地筛选出与结直肠癌抗辐射相关的潜在基因,且潜在基因在结直肠癌HCT116细胞中差异表达。
关键词: 结直肠肿瘤, 辐射抗性, 差异表达基因, 生物信息学
自噬对肿瘤放射治疗疗效的影响
路璐, 樊赛军
2019, 43(1): 61-67. doi: 10.3760/cma.j.issn.1673-4114.2019.01.011
自噬是一种重要的分解代谢过程,细胞消化和再循环自身的细胞质内容物以维持细胞稳态。自噬可以通过各种信号通路在肿瘤的发生发展中起到促进和抑制的双重作用。作为研究热点,自噬正被学者们从各个方面进行探索。然而,目前缺乏关于自噬与放疗关系的系统性总结。因此,笔者将从自噬对不同类型肿瘤放射敏感性和对放疗疗效的影响及自噬修饰用于改善肿瘤放疗疗效和预后的未来发展作一综述。
关键词: 自噬, 放射疗法, 辐射耐受性, 肿瘤
HBXIP蛋白表达对宫颈癌细胞的增殖能力及放射敏感性的影响
姜勉, 董佳丽, 李航, 樊赛军
2017, 41(5): 340-346. doi: 10.3760/cma.j.issn.1673-4114.2017.05.007
目的探讨用siRNA敲降乙肝病毒X蛋白结合蛋白(HBXIP)的表达对宫颈癌ME-180细胞的增殖能力及放射敏感性的影响。方法根据不同的处理方法,分别按两种分组方式进行分组。(1)把宫颈癌ME-180细胞分为4组:空白对照组、4 Gy γ射线照射组、HBXIP-siRNA转染组以及HBXIP-siRNA+γ射线照射联合组。采用MTT和克隆形成实验法来检测细胞增殖;采用qRTPCR检测凋亡相关蛋白Bcl-2及Bid的表达;Western blot检测蛋白激酶AKT的磷酸化水平。(2)把宫颈癌ME-180细胞分为3组:空白对照组、HBXIP-siRNA单独处理组、HBXIP-siRNA和AKT共转染组。对3组细胞进行不同剂量的γ射线照射,并采用克隆形成实验方法检测细胞生长。采用Student t-test对数据进行统计学分析,P < 0.05表示差异有统计学意义。结果 MTT实验和克隆形成实验结果显示,与γ射线照射组相比,HBXIP-siRNA转染+γ射线照射联合组的宫颈癌ME-180细胞增殖率明显降低(t=11.63、12.17,均P < 0.01),并伴随抑凋亡蛋白Bcl-2表达的降低(t=10.88,P < 0.01)和促凋亡蛋白Bid表达的增高(t=9.31,P < 0.01)。γ射线照射明显上调了HBXIP蛋白的表达水平和AKT蛋白的磷酸化水平,而转染HBXIP-siRNA则抑制了γ射线照射导致的AKT磷酸化水平的升高。另外,与HBXIP-siRNA单独处理组相比,HBXIP-siRNA和AKT共转染组中HBXIP-siRNA对宫颈癌ME-180细胞增殖的影响显著降低(t=8.96,P < 0.01)。结论降低HBXIP蛋白表达可以抑制辐照诱导的AKT磷酸化水平,进而降低宫颈癌ME-180细胞的增殖能力,同时增强其放射敏感性。
关键词: HBXIP, RNA, 小分子干扰, 宫颈肿瘤, 细胞增殖, 辐射耐受性
新型辐射增敏药RRx-001
董正川, 段玉清, 樊赛军, 李祎亮
2017, 41(3): 214-219. doi: 10.3760/cma.j.issn.1673-4114.2017.03.011
辐射增敏药可提高射线对肿瘤细胞,尤其是乏氧细胞的杀伤率,增强放疗效果,且其对有氧正常组织危害小,使用方便,因而有望成为放疗中的重要辅助药物。新型二硝基氮杂环丁烷辐射增敏药RRx-001源自航空产业。作为一氧化氮的供体分子,RRx-001可透过红细胞膜,与血红蛋白的β半胱氨酸93结合,并在乏氧环境中大量释放一氧化氮,从而提高乏氧细胞对照射的敏感度。临床实验显示RRx-001具有良好的安全性和耐受性。目前其对胆管癌、结直肠癌等肿瘤的治疗正在进行临床Ⅱ期实验。
关键词: 辐射增敏药, 一氧化氮, RRx-001
长链非编码RNA NBR2对乳腺癌细胞放射敏感性的影响
李航, 姜勉, 樊赛军
2018, 42(2): 121-128. doi: 10.3760/cma.j.issn.1673-4114.2018.02.005
目的探讨长链非编码RNA(lncRNA)BRCA1相邻基因2(NBR2)(BRCA1为乳腺癌易感基因1)对乳腺癌MCF-7和MDA-MB-231细胞放射敏感性的影响。方法根据处理方法的不同,分别按以下方式将乳腺癌细胞MCF-7和MDA-MB-231进行分组。(1)分为3组:空白对照组、4 Gy γ射线照射组和8 Gy γ射线照射组,采用实时定量PCR检测lncRNA NBR2的表达。(2)分为4组:空白对照组、NBR2转染组、4 Gy γ射线照射组以及NBR2转染+γ射线照射联合组,采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)方法来检测细胞增殖情况。(3)分为3组:空白对照组、NBR2单独转染组、NBR2+B细胞淋巴瘤2(BCL2)共转染组,对3组细胞进行不同剂量的γ射线照射,并采用MTT和克隆形成实验方法检测细胞生长情况。采用Student t-test对数据进行统计学分析,P<0.05表示差异有统计学意义。结果实时定量PCR结果显示,与空白对照组相比,4 Gy γ射线照射和8 Gy γ射线照射能够显著下调lncRNA NBR2的表达水平,差异有统计学意义(MCF-7:t=10.75、11.17,MDA-MB-231:t=11.22、12.31,均P<0.01)。MTT实验结果显示,与4 Gy γ射线照射组相比,NBR2转染+γ射线照射联合组的乳腺癌细胞增殖率明显降低,差异有统计学意义(MCF-7:t=10.55,MDA-MB-231:t=11.97,均P<0.01)。同时,lncRNA NBR2过表达可以明显下调BCL2的mRNA及蛋白表达水平。另外,与NBR2单独转染组相比,NBR2+BCL2共转染组中NBR2对细胞增殖的影响显著降低,差异有统计学意义(MCF-7:t=10.87,MDA-MB-231:t=11.37,均P<0.01)。结论辐照可以诱导lncRNA NBR2的表达水平降低,人为过表达NBR2能够抑制BCL2的蛋白表达水平,进而降低乳腺癌MCF-7和MDA-MB-231细胞的增殖能力,同时增强其放射敏感性。
关键词: RNA, 长链非编码, 乳腺肿瘤, 细胞增殖, 辐射耐受性, BRCA1相邻基因2
MiR-148a对肺癌细胞放射敏感性的影响
李航, 姜勉, 樊赛军
2018, 42(3): 248-256. doi: 10.3760/cma.j.issn.1673-4114.2018.03.010
目的探讨miR-148a对肺癌A549、H460以及H1299细胞放射敏感性的影响。方法根据不同的处理方法,分别按以下方式将肺癌A549、H460和H1299细胞进行分组。(1)将肺癌A549和H460细胞分为3组:空白对照组、4 Gy γ射线照射组和8 Gy γ射线照射组。采用实时定量PCR检测miR-148a的表达水平;(2)将肺癌A549细胞分为2组:空白对照组、miR-148a转染组;同时,将肺癌H460细胞分为2组:空白对照组、anti-miR-148a转染组。分别对转染2组细胞进行不同剂量的γ射线照射,并采用克隆形成实验方法来检测细胞增殖;(3)将肺癌A549和H1299细胞分为3组:空白对照组、miR-148a单独处理组、miR-148a和雌激素受体(ER)共转染组。分别对3组细胞进行不同剂量的γ射线照射,并采用克隆形成实验方法检测细胞生长情况。采用Student t-test对数据进行统计学分析,P < 0.05表示差异有统计学意义。结果实时定量PCR实验结果显示,γ射线照射能够显著下调肺癌A549和H460细胞中miR-148a的表达水平。克隆形成实验结果显示,与空白对照组相比,miR-148a转染能够显著增强肺癌A549细胞的放射敏感性,差异有统计学意义(t=12.16,P < 0.01),而anti-miR-148a转染能够显著降低肺癌H460细胞的放射敏感性,差异有统计学意义(t=11.93,P < 0.01)。同时,miR-148a过表达可以明显下调肺癌A549细胞中ER的mRNA及蛋白表达水平;而anti-miR-148a能够显著上调肺癌H460细胞中ER的mRNA及蛋白表达水平。另外,与miR-148a单独处理组相比,miR-148a和ER共转染组中miR-148a对肺癌A549和H1299细胞增殖的影响显著降低,差异有统计学意义(t=11.34、12.68,均P < 0.01)。结论照射可以诱导miR-148a的表达水平降低,人为过表达miR-148a能够抑制ER的蛋白表达水平,进而降低肺癌A549和H1299细胞的增殖能力,同时增强其放射敏感性。
关键词: 肺肿瘤, 辐射耐受性, MiRNA-148a, 雌激素受体, 细胞增殖
  • 首页
  • 上一页
  • 1
  • 2
  • 末页
  • 共:2页
  • 跳转
  • Go