华蟾素介导miR-497-5p/VEGFA通路调控肺癌细胞的增殖、凋亡及血管生成

目的:探讨华蟾素介导miR-497-5p/VEGFA通路对肺癌细胞增殖、凋亡及血管生成的影响。方法:选取肺癌细胞株A549、H460为研究对象,以不同浓度华蟾素（0、25、50、75、100、125、150 μg/mL）作用细胞株,采用MTT检测细胞活力。随后,设置组别为空白组、华蟾素组（100 μg/mL华蟾素）、华蟾素+inhibitor NC组、华蟾素+miR-497-5p inhibitor组,通过MTT检测细胞活力;克隆形成实验检测克隆形成能力;流式细胞术检测细胞凋亡;Western blot检测细胞中VEGF、VEGFA蛋白表达;RT-qPCR检测细胞中miR-497-5p表达;并采用荧光素酶报告实验确定miR-497-5p靶向VEGFA。结果:随着华蟾素浓度的升高,A549、H460细胞增殖活力明显下降（P＜0.01）;与空白组相比,华蟾素组A549、H460细胞克隆形成能力明显降低、细胞凋亡明显增加、VEGF和VEGFA蛋白表达明显降低、miR-497-5p mRNA表达明显升高（P＜0.01）;荧光素酶报告实验显示miR-497-5p靶向VEGFA;与华蟾素+inhibitor NC组相比,华蟾素+miR-497-5p inhibitor组A549、H460细胞中miR-497-5p mRNA表达明显降低、VEGF和VEGFA蛋白表达明显升高、细胞克隆形成能力明显增加、细胞凋亡明显被抑制、细胞增殖活力明显增加（P＜0.05）。结论:华蟾素可能通过调控miR-497-5p/VEGFA通路从而抑制肺癌细胞增殖和血管生成,并诱导细胞凋亡。     

沉默LncRNA OIP5-AS1通过上调miR-34c-5p表达增加A549R细胞放射敏感性

目的 探讨LncRNA OIP5-AS1对非小细胞肺癌放射抗拒细胞A549R的放射敏感性影响及作用机制。方法 X射线6Gy照射5次A549细胞建立放射抗拒细胞A549R。qRT-PCR检测A549、A549R细胞OIP5-AS1和miR-34c-5p表达水平。转染OIP5-AS1抑制剂或miR-34c-5p模拟物至A549R细胞,OIP5-AS1过表达质粒至A549细胞。流式细胞仪检测细胞凋亡,克隆形成实验检测细胞放射敏感性,蛋白印记检测p-Chk2和p-ATM蛋白表达水平。双荧光素酶实验验证OIP5-AS1与miR-34c-5p之间关系。结果 与A549细胞比,A549R细胞中OIP5-AS1表达上调(1.97±0.11︰1.01±0.05,P<0.05),miR-34c-5p表达下调(0.43±0.02︰1.02±0.06,P<0.05)。沉默OIP5-AS1+6Gy组A549R细胞中p-Chk2和p-ATM蛋白水平低于沉默对照+6Gy组(0.43±0.03︰1.39±0.15和0.51±0.05︰1.21±0.11,P<0.05),但凋亡率增加[(13.29±1.25)%︰(28.47±2.31)%,P<0.05]。过表达OIP5-AS1+6Gy组A549细胞中p-Chk2和p-ATM蛋白水平高于过表达对照+6Gy组(1.23±0.13︰0.75±0.06和1.08±0.11︰0.59±0.04,P<0.05)。抑制miR-34c-5p表达逆转了沉默OIP5-AS1对A549R细胞存活分数影响,增敏比为1.42。OIP5-AS1负调控miR-34c-5p表达。结论 沉默OIP5-AS1通过上调miR-34c-5p表达增强A549R细胞放射敏感性,为放疗提供潜在的靶点。     

沉默LncRNA OIP5-AS1通过上调miR-34c-5p表达增加A549R细胞放射敏感性

目的 探讨LncRNA OIP5-AS1对非小细胞肺癌放射抗拒细胞A549R的放射敏感性影响及作用机制。方法 X射线6Gy照射5次A549细胞建立放射抗拒细胞A549R。qRT-PCR检测A549、A549R细胞OIP5-AS1和miR-34c-5p表达水平。转染OIP5-AS1抑制剂或miR-34c-5p模拟物至A549R细胞,OIP5-AS1过表达质粒至A549细胞。流式细胞仪检测细胞凋亡,克隆形成实验检测细胞放射敏感性,蛋白印记检测p-Chk2和p-ATM蛋白表达水平。双荧光素酶实验验证OIP5-AS1与miR-34c-5p之间关系。结果 与A549细胞比,A549R细胞中OIP5-AS1表达上调(1.97±0.11︰1.01±0.05,P<0.05),miR-34c-5p表达下调(0.43±0.02︰1.02±0.06,P<0.05)。沉默OIP5-AS1+6Gy组A549R细胞中p-Chk2和p-ATM蛋白水平低于沉默对照+6Gy组(0.43±0.03︰1.39±0.15和0.51±0.05︰1.21±0.11,P<0.05),但凋亡率增加[(13.29±1.25)%︰(28.47±2.31)%,P<0.05]。过表达OIP5-AS1+6Gy组A549细胞中p-Chk2和p-ATM蛋白水平高于过表达对照+6Gy组(1.23±0.13︰0.75±0.06和1.08±0.11︰0.59±0.04,P<0.05)。抑制miR-34c-5p表达逆转了沉默OIP5-AS1对A549R细胞存活分数影响,增敏比为1.42。OIP5-AS1负调控miR-34c-5p表达。结论 沉默OIP5-AS1通过上调miR-34c-5p表达增强A549R细胞放射敏感性,为放疗提供潜在的靶点。     

miR-339-5p通过靶向调节p53表达影响脑胶质瘤细胞的转移活性分析

目的 研究miR-339-5p对脑胶质瘤U87细胞的影响,并探讨相关机制。方法 将U87细胞分为4组：空白组、miR-339-5p mimic组、si-p53组、miR-339-5p mimic+si-p53组。通过MTT实验检测细胞增殖率;通过Transwell实验检测U87细胞侵袭能力;通过流式细胞术检测细胞凋亡率;通过荧光素酶报告基因实验检测miR-339-5p与p53的靶向关系;通过实时荧光定量聚合酶链反应和蛋白免疫印迹分析miR-339-5p和p53的表达水平。结果 miR-339-5p mimic组和miR-339-5p mimic+si-p53组细胞的miR-339-5p表达水平高于空白组和si-p53组,miR-339-5p mimic组细胞的p53 mRNA和蛋白水平均高于空白组、si-p53组和miR-339-5p mimic+si-p53组(P<0.05)。miR-339-5p mimic和p53-WT共转染的细胞荧光素酶活性明显增加(P<0.05)。miR-339-5p mimic组细胞增殖率和细胞侵袭数量低于空白组、si-p53组和miR-339-...     

miR-125a-5p通过靶向APAF1 增强非小细胞肺癌细胞吉非替尼耐药性

目的：探讨miR-125a-5p 在诱导非小细胞肺癌（non-small cell lung cancer，NSCLC）细胞吉非替尼（gefitinib，Gef）耐药中的作用及其机制。方法：选用人NSCLC 耐药细胞株A549/GR 和NSCLC细胞株A549，将miR-125a-5p mimic、miR-125a-5p inhibitor、pcDNA3.1-APAF1、空载体pcDNA3.1 转染至A549/GR细胞。用qPCR检测细胞中miR-125a-5p 的表达水平，用MTT法、Transwell 小室法和流式细胞术检测Gef 对细胞增殖、迁移和凋亡的影响。用双荧光素酶报告基因实验验证miR-125a-5p 与细胞凋亡蛋白酶活化因子1（apoptotic peptidase activating factor 1，APAF1）的靶向关系，用Western blotting检测A549/GR细胞中APAF1蛋白水平，用比色法测定细胞中caspase-3 及caspase-9 表达水平。结果：A549/GR 细胞中miR-125a-5p 表达水平显著高于A549 细胞（P<0.01）。敲降miR-125a-5p 显著增强Gef 对A549/GR细胞增殖、迁移的抑制作用（均P<0.05），并促进细胞凋亡（P<0.01）。双荧光素酶报告基因实验证实miR-125a-5p 靶向APAF1，并负调控其表达。进一步实验显示，miR-125a-5p 通过靶向下调APAF1 缓解Gef 对A549/G 细胞增殖、迁移的抑制作用及凋亡的促进作用（均P<0.05），减弱Gef引起的凋亡相关蛋白caspase-3及caspase-9表达的上调（均P<0.05）。结论：miR-125a-5p 促进NSCLC细胞Gef 耐药，其机制是通过靶向APAF1 而促进细胞的增殖、迁移并抑制凋亡。     

miR-361-5p对肾细胞癌ACHN细胞恶性生物学行为的影响

目的：探讨miR-361-5p对肾细胞癌ACHN细胞增殖、侵袭、迁移、凋亡及其细胞周期的影响。方法：将miR-361-5p mimics和miR-361-5p inhibitor分别转染至肾癌ACHN细胞中,用qPCR检测转染细胞中miR-361-5p的表达水平,用MTT法、划痕愈合实验、Transwell实验、流式细胞术分别检测细胞的增殖、迁移、侵袭、细胞周期和凋亡水平。结果：与空白对照组和 Mimics-NC组比较,miR-361-5p mimics组ACHN细胞中miR-361-5p表达水平显著升高（P<0.01）,细胞的增殖、侵袭和迁移能力均显著减弱（均P<0.01）,而凋亡率升高（P<0.01）。与空白对照组或Inhibitor-NC组比较,miR-361-5p inhibitor组细胞中miR-361-5p表达水平显著下降（P<0.01）,细胞的增殖、侵袭和迁移能力均增强（均P<0.01）,细胞周期运转加速（P<0.01）,凋亡率降低（P<0.05）。结论：miR-361-5p可抑制肾癌ACHN细胞的增殖、侵袭和迁移,并诱导细胞凋亡,其在肾癌发生发展过程中发挥重要的抑制作用。     

lncRNA MEG3通过下调miR-7-5p表达对鼻咽癌细胞放射敏感性的影响

目的 探讨lncRNA MEG3对鼻咽癌细胞的放射敏感性影响及潜在作用机制。方法 实验设置过表达对照组、过表达MEG3组、抑制miR-NC组、抑制miR-7-5p组、过表达对照+4 Gy组、过表达MEG3+4 Gy组、抑制miR-NC+4 Gy组、抑制miR-7-5p+4 Gy组、过表达MEG3+过表达miR-NC组、过表达MEG3+过表达miR-7-5p组。采用qRT-PCR检测miR-7-5p和MEG3表达;细胞克隆形成实验检测鼻咽癌细胞放射敏感性;流式细胞术检测细胞凋亡;双荧光素酶报告基因检测实验检测荧光活性。结果 MEG3在鼻咽癌组织和细胞系中低表达;过表达MEG3和抑制miR-7-5p表达可增加鼻咽癌细胞放射敏感性且促进细胞凋亡。MEG3可靶向调节miR-7-5p表达;过表达miR-7-5p逆转了过表达MEG3对鼻咽癌细胞放射增敏和促进细胞凋亡的作用。结论 过表达MEG3增加鼻咽癌细胞放射敏感性,并促进细胞凋亡,其机制可能与下调miR-7-5p有关。     

lncRNA MEG3通过下调miR-7-5p表达对鼻咽癌细胞放射敏感性的影响

目的 探讨lncRNA MEG3对鼻咽癌细胞的放射敏感性影响及潜在作用机制。方法 实验设置过表达对照组、过表达MEG3组、抑制miR-NC组、抑制miR-7-5p组、过表达对照+4 Gy组、过表达MEG3+4 Gy组、抑制miR-NC+4 Gy组、抑制miR-7-5p+4 Gy组、过表达MEG3+过表达miR-NC组、过表达MEG3+过表达miR-7-5p组。采用qRT-PCR检测miR-7-5p和MEG3表达;细胞克隆形成实验检测鼻咽癌细胞放射敏感性;流式细胞术检测细胞凋亡;双荧光素酶报告基因检测实验检测荧光活性。结果 MEG3在鼻咽癌组织和细胞系中低表达;过表达MEG3和抑制miR-7-5p表达可增加鼻咽癌细胞放射敏感性且促进细胞凋亡。MEG3可靶向调节miR-7-5p表达;过表达miR-7-5p逆转了过表达MEG3对鼻咽癌细胞放射增敏和促进细胞凋亡的作用。结论 过表达MEG3增加鼻咽癌细胞放射敏感性,并促进细胞凋亡,其机制可能与下调miR-7-5p有关。     

miR-379-5p靶向CTSL调控肺癌细胞增殖、迁移、侵袭及凋亡的分子机制

目的:探讨微小RNA-379-5p（miR-379-5p）通过靶向组织蛋白酶L(CTSL)调控肺癌细胞增殖、迁移、侵袭及凋亡的作用机制。方法:选择本院收治的肺癌患者57例,取患者肺癌组织与癌旁组织,应用qRT-PCR法检测miR-379-5p与CTSL mRNA的表达水平;免疫组化法检测CTSL蛋白的表达情况。miR-con（miR-con组）、miR-379-5p mimics（miR-379-5p组）分别转染肺癌NC-H446细胞,miR-379-5p mimics与pcDNA（miR-379-5p+pcDNA组）、miR-379-5p mimics与pcDNA-CTSL（miR-379-5p+pcDNA-CTSL组）分别共转染肺癌NC-H446细胞,MTT检测细胞增殖情况;流式细胞术检测细胞凋亡率;Transwell迁移与侵袭实验检测细胞迁移与侵袭能力。双荧光素酶报告实验验证miR-379-5p与CTSL之间的靶向关系。Western blot检测CTSL、Cyclin D1、MMP-2、MMP-9、Cleaved caspase-3蛋白的表达。结果:肺癌组织中miR-379-5p的表达水平显著低于癌旁组织（P＜0.05）,而CTSL mRNA及蛋白阳性率均显著高于癌旁组织（P＜0.05）;与miR-con组比较,miR-379-5p组细胞增殖、迁移、侵袭能力显著降低（P＜0.05）,细胞凋亡率显著增加（P＜0.05）,明显抑制Cyclin D1、MMP-2、MMP-9蛋白表达（P＜0.05）,而促进Cleaved caspase-3蛋白表达（P＜0.05）;双荧光素酶报告实验证明miR-379-5p可负向调控CTSL表达与活性;CTSL过表达可逆转miR-379-5p过表达对肺癌细胞增殖、迁移、侵袭及凋亡的调控作用。结论:miR-379-5p过表达通过靶向CTSL而减弱肺癌细胞增殖、迁移及侵袭能力,诱导细胞凋亡。     

circ_0001429 靶向miR-139-5p/TGIF1 分子轴调控膀胱癌T24 细胞恶行生物学行为

目的：探究circ_0001429 通过调控miR-139-5p/ 转录生长因子影响因子1（TGF-interacting factor 1,TGIF1）分子轴对膀胱癌T24 细胞恶性生物学行为的影响。方法：采用qPCR实验检测circ_0001429 在膀胱癌细胞系SW780、T24、5637 和人膀胱上皮永生化细胞SV-HUC-1 中的表达水平,双荧光素酶报告基因实验验证miR-139-5p、circ_0001429 与TGIF1 之间的靶向调控关系;将T24 细胞分为NC组、sh-circ_0001429 组、miR-139-5p mimics 组、sh-TGIF1 组、pcDNA-circ_0001429+sh-TGIF1 组、miR-139-5p mimics+pcDNA-TGIF1 组以及sh-circ_0001429+miR-139-5p inhibitor 组,Western blotting 检测各组细胞中TGIF1 的表达水平,CCK-8 法、Transwell 实验和流式细胞术分别检测circ_0001429、miR-139-5p 和TGIF1 对T24 细胞增殖、侵袭、迁移和凋亡的影响。结果：circ_0001429 在3 珠膀胱癌细胞系中呈高表达（均P<0.01）,敲降circ_0001429 可以显著抑制T24 细胞增殖、侵袭、迁移并促进细胞凋亡（P<0.05 或P<0.01）;双荧光素酶报告基因验证结果显示,circ_0001429 与miR-139-5p、miR-139-5p 与TGIF1 存在靶向关系;过表达miR-139-5p 显著抑制T24 细胞增殖、侵袭、迁移并促进细胞凋亡（均P<0.01）。回复实验进一步证实circ_0001429 和TGIF1 竞争性结合miR-139-5p 促进T24 细胞增殖、侵袭、迁移且抑制细胞凋亡（均P<0.01）。结论：circ_0001429 与TGIF1 竞争结合miR-139-5p 促进膀胱癌细胞T24 的增殖、侵袭、迁移且抑制细胞凋亡。     

Comparative Antitumor Activity and Intestinal Toxicity of 5'-Deoxy-5-fluorouridine and Its Prodrug Trimethoxybenzoyl-5'-deoxy-5-fluorocytidine

N⁴-Trimethoxybenzoyl-5'-deoxy-5-fluorocytidine (Ro 09–1390), a prodrug of the cytostatic 5'-deoxy-5-fluorouridine (5'-DFUR), was synthesized with the aim of reducing of the dose-limiting toxicity of 5'-DFUR, which is diarrhea. In mice bearing Lewis lung carcinoma, 5'-DFUR given po produced a substantial amount of 5-fluorouracil (5-FU) in the intestinal tract as well as in tumors, where the enzyme pyrimidine nucleoside phosphorylase, essential for conversion of 5'-DFUR to 5-FU, is predominantly located. With the oral administration of Ro 09–1390 only a small amount of 5-FU was formed in the intestine; however, the administration of Ro 09–1390 and 5'-DFUR at the same dose produced similar amounts of 5-FU in tumor tissues. These differences in metabolism were reflected in their toxicity and antitumor efficacy. The administration of 5'-DFUR resulted in damage to the intestinal mucosal membrane and diarrhea in normal mice, whereas Ro 09–1390 was much less toxic to the intestinal tract. As regards antitumor activity, Ro 09–1390 and 5'-DFUR at equivalent doses inhibited the growth of Lewis lung carcinoma to similar extents. Since Ro 09–1390 was much less toxic to the intestinal tract than 5'-DFUR, mice bearing Lewis lung carcinoma could be given Ro 09–1390 daily over a longer period and at a higher dose, resulting in a longer survival time.     

Down-regulation of KLF5 in cancer-associated fibroblasts inhibit gastric cancer cells progression by CCL5/CCR5 axis

It was well known that cancer-associated fibroblasts (CAFs) were an essential factor in tumor progression. However, the actual mechanism of stromal fibroblasts activation and tumor promoting effects remain unclear. Here, we showed that KLF5 expression was more frequently observed in gastric cancer-associated fibroblasts compared with normal mucosal fibroblasts. Moreover, KLF5 expression in tumor stroma was closely associated with clinicopathological features such as tumor size, invasion depth, cell grade and lymph node metastasis, as well as poor prognosis in patients with gastric cancer. In addition, we further demonstrated that KLF5-regulating CAFs affect gastric cancer cells progression by CCL5 secretion and activation of CCR5. Taken together, we concluded that KLF5 expression in gastric cancer-associated fibroblasts contribute to poor survival and promote cancer cells progression by activation of CCL5/CCR5 axis, which suggesting that KLF5 in CAFs might be considered as a promising target for the treatment of gastric cancer.     

Comparative Antitumor Activity of 5-Fluorouracil and 5'-Deoxy-5-fluorouridine in Combination with Radiation Therapy in Mice Bearing Colon 26 Adcnocarcinoma

The present study compared the antitumor activities of chemotherapy with 5-fluorouracil (5-FU) and with its prodrug 5'-deoxy-5-fluorouridine (5'-DFUR) in combination with radiotherapy on a solid colon 26 adenocarcinoma in the mouse. A single administration of 5'-DFUR immediately after local irradiation on day 10 after tumor inoculation produced more than additive antitumor effects, while only an additive effect was observed in the combined treatment with 5-FU and radiation. This over-additive effect of 5'-DFUR was more obvious in a fractionated-dose treatment schedule, where the same combined modality treatment was given three times on days 6, 10 and 14 after inoculation of the tumor cells. 5'-DFUR enhanced the radiation effects on the tumor in terms of the delay in tumor growth as well as the increase in the survival time. 5-FU produced only a marginal additive antitumor effect. Furthermore, radiation damage to normal tissues (skin damage by local irradiation and bone marrow and spleen damage by whole-body irradiation) was not enhanced by 5'-DFUR, though radiation damage to the thymus was additive. On the other hand, 5-FU produced toxic effects that were additive for all normal tissues tested. Thus, at doses that were the most effective against tumors, relative therapeutic gain factors (the ratio of the effect on tumors to that on the bone marrow) of 5'-DFUR and 5-FU were 1.24 and 0.49, respectively. These results suggest that 5'-DFUR will have a greater potential than 5-FU in combined modality treatment of cancer patients.     

Effect of 5-Fluorocytosine and 5-Fluorouracil on Human and Rat Hepatic Cytochrome P 450 Die Wirkung von 5-Fluorcytosin und 5-Fluoruracil auf Leber-Cytochrom-P 450 in Mensch und Ratte

Hepatotoxicity is a well-known side effect of the antifungal drug 5-fluorocytosine. The underlying mechanisms of this toxicity are unknown. The present in vitro study was, therefore, designed to assess the influence of 5-fluorocytosine and 5-fluorouracil on the hepatic cytochrome P 450 concentration in human and rat liver microsomes. Incubation of human or rat hepatic microsomes for 1 h with 5-fluorocytosine up to 500 micrograms/ml or with 5-fluorouracil up to 200 micrograms/ml did not influence the cytochrome P 450 concentration. In comparison the amount of cytochrome P 450 in human liver, however, was lower than in rat liver microsomes and a more interindividual variation was observed.     

Wnt5a在肿瘤研究中的进展

近年来研究发现WnL5a与肿瘤发生、发展关系密切.根据细胞类型不同,Wnt5a既可以起到促癌作用,又可以起到抑癌作用,其信号级联反应复杂,受多种因子调节.     

Wnt5a在肿瘤研究中的进展

近年来研究发现WnL5a与肿瘤发生、发展关系密切.根据细胞类型不同,Wnt5a既可以起到促癌作用,又可以起到抑癌作用,其信号级联反应复杂,受多种因子调节.     

Wnt5a在肿瘤研究中的进展

近年来研究发现WnL5a与肿瘤发生、发展关系密切.根据细胞类型不同,Wnt5a既可以起到促癌作用,又可以起到抑癌作用,其信号级联反应复杂,受多种因子调节.     

Wnt5a在肿瘤研究中的进展

近年来研究发现WnL5a与肿瘤发生、发展关系密切.根据细胞类型不同,Wnt5a既可以起到促癌作用,又可以起到抑癌作用,其信号级联反应复杂,受多种因子调节.     

Wnt5a在肿瘤研究中的进展

近年来研究发现WnL5a与肿瘤发生、发展关系密切.根据细胞类型不同,Wnt5a既可以起到促癌作用,又可以起到抑癌作用,其信号级联反应复杂,受多种因子调节.