ZNRD1 might mediate UV irradiation related DNA damage and repair in human esophageal cancer cells by regulation of ERCC1

The downregulation of zinc ribbon domain‐containing 1 (ZNRD1) protein was recently found to partially reverse the resistance of human leukemia cells toward chemical therapeutic drugs. Therefore, the ZNRD1 protein might be involved in the process of DNA damage and repair. To explore the possible protective effects of ZNRD1 on DNA damage induced by ultraviolet (UV)‐C irradiation in human esophageal squamous cancer cell line EC109, we designed and transfected a expression vector into EC109 cells, and established an overexpression cell line. The single‐cell gel electrophoresis (comet assay) was used to investigate the DNA damage and repair in UV‐C‐irradiated control and transfected cells. It was found that the ZNRD1‐expressing cells exhibited a significant enhanced DNA repair capacity. Moreover, the overexpression of ZNRD1 could upregulate the expression of excision repair cross‐complementing 1 (ERCC1) gene. Collectively, these findings suggested that ZNRD1 might play an important role in the process of DNA damage and repair by regulating the expression of ERCC1.     

Significance of perivascular tumour cells defined by CD109 expression in progression of glioma

In the progression of glioma, tumour cells often exploit the perivascular microenvironment to promote their survival and resistance to conventional therapies. Some of these cells are considered to be brain tumour stem cells (BTSCs); however, the molecular nature of perivascular tumour cells has not been specifically clarified because of the complexity of glioma. Here, we identified CD109, a glycosylphosphatidylinositol‐anchored protein and regulator of multiple signalling pathways, as a critical regulator of the progression of lower‐grade glioma (World Health Organization grade II/III) by clinicopathological and whole‐genome sequencing analysis of tissues from human glioma. The importance of CD109‐positive perivascular tumour cells was confirmed not only in human lower‐grade glioma tissues but also in a mouse model that recapitulated human glioma. Intriguingly, BTSCs isolated from mouse glioma expressed high levels of CD109. CD109‐positive BTSCs exerted a proliferative effect on differentiated glioma cells treated with temozolomide. These data reveal the significance of tumour cells that populate perivascular regions during glioma progression, and indicate that CD109 is a potential therapeutic target for the disease. Copyright © 2017 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

食管鳞癌中Klf17蛋白表达及其对Eca109细胞迁移和侵袭能力的影响

目的探讨KLF17与临床病理特征的关系;探讨KLF17对Eca109细胞迁移和侵袭能力的影响及其作用机制。方法采用免疫组化法,检测KLF17在食管鳞癌及癌旁正常食管上皮中的表达;构建KLF17慢病毒过表达载体感染Eca109细胞,细胞分组为转染组(CON)、空载体转染组(Ad-GFP)和慢病毒转染组(Ad-KLF17)。Real timePCR检测KLF17 mRNA的表达、Western blot检测KLF17、E-cadherin和N-cadherin蛋白的表达;细胞划痕实验及体外Transwell实验观察Eca109细胞迁移和侵袭能力。结果 KLF17蛋白表达与侵袭深度、TNM分期显著相关(P0.05);real time-PCR及Western blot结果显示Ad-KLF17组KLF17及上皮标志物E-cadherin表达明显高于Ad-GFP组及对照组(P0.05),而间质标志物N-cadherin表达明显低于Ad-GFP组及对照组(P0.05);划痕实验与体外Transwell实验结果显示Ad-KLF17组细胞迁移距离及细胞数量明显短于和少于Ad-GFP组和对照组(P0.05)。结论 KLF17能抑制Eca109细胞发生上皮-间质转化,与其迁移、侵袭能力相关。     

MKK6过表达调控SIRT1影响食管癌细胞肿瘤行为学的体外研究

目的 向食管癌Eca109细胞中转染pcDNA3.1（+）/myc-His A-丝裂原活化蛋白激酶6（MKK6）重组过表达载体,观察过表达MKK6影响沉默信息转录调控因子（SIRT1）表达后肿瘤细胞增殖、凋亡以及侵袭力发生的变化,探讨p38-MAPK-SIRT1调节轴的存在性及其对Eca109细胞各项肿瘤行为学造成的影响。 方法 根据GenBank中MKK6基因的mRNA序列设计引物,构建MKK6过表达载体;将Eca109细胞分空载体转染组、MKK6过表达载体转染组、MKK6-SIRT1 ShRNA共转染组和MKK6-RES（白藜芦醇）组,相应处理后,采用Western blot测定各组Eca109细胞的SIRT1及MKK6表达水平,MTT法检测各组Eca109细胞的增殖活力,Transwell法观察各组Eca109细胞的侵袭力,流式细胞术检测各组Eca109细胞的凋亡情况。 结果 测序结果表明成功构建MKK6过表达载体;转染MKK6过表达载体可降低Eca109细胞的内源性SIRT1表达,减弱Eca109细胞的增殖活力,促其凋亡,同时可使Eca109细胞的侵袭力增强,以上各项改变均与SIRT1的表达水平有关。 结论 Eca109细胞内可能存在p38-MAPK-SIRT1调节轴,MKK6及其下游因子对SIRT1可能具有反馈调节作用,可引起Eca109细胞各项肿瘤行为学的变化。     

ｐ２７Ｖａｌ１０９Ｇｌｙ基因多态性在肿瘤发生发展中的作用机制

ｐ２７是细胞周期蛋白依赖性激酶抑制剂（ＣＤＫＩ）家族中的重要成员,能阻滞细胞进入Ｇ１期或Ｓ期,其蛋白表达水平及活性改变与肿瘤发生发展有关。ｐ２７基因存在多个单核苷酸多态性（ＳＮＰｓ）位点,其中ｐ２７Ｖａｌ１０９Ｇｌｙ（ｒｓ２０６６８２７）基因多态性位于其降解蛋白Ｊａｂ１结合区,导致ｐ２７与降解蛋白Ｊａｂ１的不同结合活性,从而影响其在肿瘤发生发展中的作用,因此被认为是ｐ２７基因最重要的多态性。本文拟综述ｐ２７Ｖａｌ１０９Ｇｌｙ基因多态性在肿瘤发生发展中的分子机制。     

RNA干扰HIF-1α对Eca109食管癌细胞放疗敏感性的影响

目的探讨RNA干扰乏氧诱导因子1α(HIF-1α)对Eca109食管癌细胞放疗敏感性的影响。方法构建干扰HIF-1α质粒pSilencer 2.1 HIF-1α,采用实时荧光定量Real-time PCR、Western Blot方法验证干扰效果,通过MTT、流式细胞仪和克隆形成试验观察RNA干扰HIF-1α对Eca109食管癌细胞的放疗敏感性的影响。结果 RNA干扰可抑制Eca109细胞中HIF-1α和VEGF mRNA转录和蛋白的表达;HIF-1α基因沉默后细胞凋亡增加,且肿瘤细胞对放疗的敏感性增加。结论靶向HIF-1α的shRNA能有效抑制Eca109食管癌细胞的HIF-1α和VEGF基因表达,促进细胞凋亡,增加肿瘤细胞的放疗敏感性。     

脂质体槲皮素对食管癌细胞增殖及蛋白表达的影响及机制

目的比较不同脂质体槲皮素对人食管癌Eca109和Eca9706细胞增殖的作用,并初步探讨其作用机制。方法采用旋转蒸发法制备氯仿和甲醇溶解类脂物质的脂质体槲皮素LQ1和按同比例的氯仿和DMSO作为溶剂的脂质体槲皮素LQ2,用DMSO直接溶解槲皮素制备非脂质体槲皮素nLQ,分别作用于Eca109和Eca9706细胞（分别作为LQ1、LQ2及nLQ组）,同时设立对照组。MTT实验检测各组细胞抑制率,免疫组织化学染色和免疫印迹法检测磷酸酶基因（PTEN）和细胞周期素D1（Cychn D1）蛋白表达。结果各组细胞增殖的抑制效应、上调PTEN和下调Cyclin D1蛋白表达的效应呈现同一趋势,即LQ2组〉LQ1组〉nLQ组〉对照组,P〈0．05。结论脂质体槲皮素对食管癌细胞增殖有抑制作用,LQ2的抑制率高于LQ1;上调PTEN表达、下调Cyclin D1表达可能为其作用机制之一。