The histone deacetylase inhibitor, 2-propylpentanoic acid, increases the chemosensitivity and radiosensitivity of human glioma cell lines in vitro

Background Treatment for malignant glioma generally consists of cytoreductive surgery followed by radiotherapy and chemotherapy. In this study, we intend to investigate the effects of 2-propylpentanoic acid (VPA), a histone deacetylase inhibitor, on chemosensitivity and radiosensitivity in human glioma cell lines. Methods Human glioma cell lines, T98-G, and SF295, were treated with temozolomide (TMZ) or irradiation (IR), with or without VPA (1.0 mmol/L). Then, cytotoxicity and clonogenic survival assay was performed. Cell cycle stage, apoptosis, and autophagy were also detected using flow cytometry and dansyl monocadaverin (MDC) incorporation assay. One-way analysis of variance (ANOVA) and t-test were used to analyze the differences among variant groups. Results Mild cytotoxicity of VPA was revealed in both cell lines, T98-G and SF295, with the 50% inhibiting concentration (IC50) value of (3.85?0.58) mmol/L and (2.15?0.38) mmol/L, respectively; while the IC50 value of TMZ was (0.20?0.09) mmol/L for T98-G and (0.08?0.02) mmol/L for SF295. Moreover, if combined with VPA (1.0 mmol/L) for 96 hours, the sensitivity of glioma cells to TMZ was significant increased (P <0.05). The surviving fractions at 2 Gy (SF2) of T98-G and SF295 cells exposed to IR alone were 0.52 and 0.58. However, when VPA was combined with IR, the SF2 of T98-G and SF295 dropped to 0.39 (P=0.047) and 0.49 (P=0.049), respectively. Treatment with VPA plus TMZ or IR also resulted in a significant decrease in the proportion of cells in the G2 phase and increased apoptotic rates as well as autophagy in T98-G and SF295 cell lines (P <0.01). Conclusion VPA may enhance the activities of TMZ and IR on glioma cells possibly through cell cycle block and promote autophagy, and thus could be a potential sensitizer of glioma treatment.     

丙戊酸钠诱导人肺癌细胞MICA表达上调增强NK细胞对其杀伤作用的研究

目的 观察丙戊酸钠(valproic acid,VPA)对人肺癌细胞A549、SP-CA-1、NCIH446中的MHCⅠ类链相关基因A(MICA)表达的影响,并比较肺癌细胞经VPA处理前后NK细胞对其杀伤作用的差异.方法 将0.75～12.0 mmol/L VPA作用于A549、SP-CA-1、NCIH446细胞,与未加VPA的对照组进行比较,观察VPA对人肺癌细胞生长的影响.选择对细胞生长无影响的1.50 mmol/L和3.00 mmol/L VPA诱导3株肺癌细胞4 d,通过RT-PCR反应和免疫荧光法分别检测MICA在mRNA和蛋白质水平的变化.LDH法检测VPA处理肺癌细胞前后其被NK细胞杀伤程度的变化.结果 ＞6.0 mmol/L VPA对3株肺癌细胞均表现出抑制其生长.用1.50mmol/L和3.00 mmol/L VPA诱导4 d后的3株肺癌细胞,与对照组比较,其MICA的mRNA转录水平和蛋白质表达提高(P＜0.05);1.50 mmol/L和3.00 mmol/L VPA诱导表达MICA后的肺癌细胞均比未经VPA处理的肺癌细胞被NK杀伤的程度高(P＜0.05).3.00 mmol/L VPA浓度组3株肺癌细胞的MICA mRNA、蛋白质表达水平及其被NK细胞杀伤的程度均高于1.50 mmol/L VPA浓度组(P＜0.05).结论 VPA通过上调MICA抗肺癌作用的新机制,为肺癌临床免疫生物治疗提供了实验依据.     

Inhibition of Ciliogenesis Enhances the Cellular Sensitivity to Temozolomide and Ionizing Radiation in Human Glioblastoma Cells

Objective To investigate the function of primary cilia in regulating the cellular response to temozolomide(TMZ) and ionizing radiation(IR) in glioblastoma(GBM).Methods GBM cells were treated with TMZ or X-ray/carbon ion. The primary cilia were examined by immunostaining with Arl13 b and γ-tubulin, and the cellular resistance ability was measured by cell viability assay or survival fraction assay. Combining with cilia ablation by IFT88 depletion or chloral hydrate and induction by lithium chlorid...     

siRNA干扰微管微丝交连因子1表达增强替莫唑胺诱导的胶质瘤细胞毒性

目的 研究微管微丝交连因子1(MACF1)在胶质母细胞瘤应对TMZ刺激中的作用.方法 替莫唑胺刺激人类胶质母细胞瘤细胞系U87细胞后,通过Western blot、RT-PCR和免疫荧光检测其蛋白表达和细胞内定位.通过RNA干扰技术干扰MACF1的表达.通过裸鼠皮下成瘤和免疫组化检测在活体中TMZ刺激后胶质母细胞瘤中MACF1的表达.结果 在体外培养和体内成瘤中均发现替莫唑胺刺激后胶质母细胞瘤MACF1的表达上调(差异约2倍),胞内分布改变(P＜0.01).敲除MACF1表达的胶质母细胞瘤细胞在TMZ刺激后其增殖能力下调约45％(P＜0.01).替莫唑胺诱导胶质母细胞瘤MACF1表达上调的同时,其细胞骨架发生重组.结论 MACF1可能成为胶质母细胞瘤治疗的一个潜在靶点.     

A 2-way cross-over, open-labeled trial to compare efficacy and safety of insulin Aspart and Novolin R delivered with CSII in 21 Chinese diabetic patients

Background Subcutaneous absorption is accelerated by the monomeric conformation of insulin Aspart, which provides good glycemic control with a lower risk of hypoglycemia and less body weight increase. In the present study we investigated the efficacy and safety of a rapid-acting human insulin analogue （insulin Aspart） delivered with continuous subcutaneous insulin infusion （CSII） into Chinese diabetic patients. Methods A total of 21 patients with type 1 or type 2 diabetes were recruited for the 2-way cross-over, open-labeled trial, and then randomized to Group A （n=-10, treated with insulin Aspart） or Group B （n=11, treated with Novolin R）. Insulin Aspart and Novolin R were administered by CSII. Capillary glucose concentrations were measured at 8 time points, pre-prandial and postprandial, bedtime （10 pm）, midnight （2 am） every day during the treatment. Results The average capillary glucose profiles for the day were much better controlled in Group A than in Group B （P〈0.01）. The blood glucose levels were particularly better controlled in Group A than in Group B at pre-breakfast （（6.72±1.24） mmol/L vs （7.84±1.58） mmol/L, P=0.014）, post-breakfast （（8.96±2.41） mmol/L vs （11.70±3.11） mmol/L, P=0.0028）, post-supper （（8.15±2.10） mmol/L vs （10.07±2.36） mmol/L, P=0.008）, bed time （（7.73±1.72） mmol/L vs （9.39±.2.05） mmol/L, P=0.007） and midnight （（6.32±1.16） mmol/L vs （7.48±1.36） mmol/L, P=0.0049）. There was no significant difference in the frequency of hypoglycemic episodes between the two groups. Conclusion Insulin Aspart results in better control of blood glucose levels than regular human insulin （Novolin R） in diabetic patients during delivery by CSII.     

丙戊酸钠联合全反式维甲酸促进子宫颈癌细胞衰老的发生

目的：研究丙戊酸钠( VPA)联合全反式维甲酸( AT-RA)诱导子宫颈癌细胞衰老作用,并探讨其分子机制。方法实验分为对照组、VPA组、ATRA组、VPA+ ATRA组,采用3 mmol/L VPA、1μmol/L ATRA单独或联合作用于人子宫颈癌HeLa及 SiHa细胞,对照组仅加溶媒;采用 CellTiter 96? AQueous 法检测细胞增殖;β-半乳糖苷酶化学染色法检测细胞衰老;Q-PCR和Western blot法分别检测衰老相关基因P16、P63、hTERT的mRNA和蛋白表达变化。结果 VPA对HeLa及SiHa细胞均有生长抑制作用,与ATRA联合抑制效果明显优于单独用药( P <0．05);β-半乳糖苷酶染色显示,VPA+ATRA 组衰老细胞比例显著增加,与对照组及VPA组、ATRA 组比较,差异有统计学意义( P <0．05);Q-PCR和Western blot法结果显示,VPA单独和联合ATRA可上调P16、P63的表达,降低hTERT的表达,且联合用药优于单独用药及对照组( P<0．05)。结论 VPA联合ATRA可抑制子宫颈癌细胞增殖,上调 P16和 P63的表达,下调hTERT的表达,可能是其诱导细胞衰老的分子机制。     

Growth of G422 glioma implanted in the mouse brain was affected by the immune ability of the host

Background  It is generally accepted that gliomas are the most common primary brain tumors with poor prognosis. We aimed to explore the relationship of the immunity of the central nervous system and the genesis and development of glioma.

Methods  G422 glioma was implanted in the brain of BALB/c mice (immuno-competent mice), nude mice (T cell related immuno-deficient) and complement C3 knock-out mice (complement C3 related immunodeficient). The survival time of the host, growth and histopathology of the tumor, and concentrations of tumor necrosis factor-α (TNF-α) and interferon-γ (INF-γ) in tumor tissues were assessed.

Results  Tumor spheres were formed in all mice after injection, and glial fibrillary acidic protein (GFAP) positive staining of the cells declared their glioma origin. The longest median survival time of (44.3±6.0) days was found in BALB/c mice, followed by (24.8±5.2) days in nude mice and the shortest (18.6±5.8) days in complement C3 knock-out mice. Accordingly, the growth of the tumor was fastest in complement C3 knock-out mice, followed by the nude mice and slowest in the BALB/c mice. Although the proportions of infiltrating CD68⁺ lymphocytes in tumor tissues showed no significant difference (P >0.05), TNF-α level in the nude and C3 knock-out mice, (28.11±4.86) μmol/L and (22.87±6.36) μmol/L respectively, were significantly lower (P <0.01) than that in the BALB/c mice, which was (230.21±39.17) μmol/L. The INF-γ level was highest in the BALB/c mice ((180.76±29.19) μmol/L), followed by the nude mice ((113.46±23.76) μmol/L) and then the C3 knock-out mice ((16.84±4.45) μmol/L). 

Conclusions  The G422 glioma implanted in the brains of mice with different immune ability would be a useful model for studying the relationship of the immune system and tumor in the central nervous system. Furthermore, the T cells and complement C3 compartments of the immune response may affect the growth of implanted tumors and inflammatory factors such as TNF-α and INF-γ.

     

2型糖尿病患者血清铁蛋白、转铁蛋白的水平及与胰岛素抵抗的相关性分析

目的分析2型糖尿病（T2DM）患者血清铁蛋白（SF）、转铁蛋白（TRF）水平及与胰岛素抵抗的相关性。方法选择山西医科大学第一医院T2DM患者60例为T2DM组,选择同期健康体检者56例为对照组,比较两组收缩压（SBP）、舒张压（DBP）、体重指数（BMI）、总胆固醇（TC）、尿酸（UA）、腰臀比（WHR）、三酰甘油（TG）、空腹血糖（FPG）、空腹胰岛素（FINS）、胰岛素抵抗指数（HOMA-IR）、高密度脂蛋白胆固醇（HDL-C）、胰岛β细胞功能（HOMAβ-cell）、血清铁蛋白（SF）、转铁蛋白（TRF）;分析SF、TRF的相关影响因素。结果①两组DBP、TC、UA水平差异无统计学意义（P〉0.05）;T2DM组SBP[（133.95±17.64）mm Hg]、BMI[（24.57±3.04）kg/m2]、WHR（0.91±0.04）、TG[（2.09±0.87）mmol/L]、FPG[（7.90±3.04）mmol/L]、FINS[（13.38±6.27）mU/L]、HOMA-IR[（4.64±2.02）]等指标高于对照组[（125.02±12.56）mm Hg、（23.02±2.93）kg/m2、（0.86±0.05）、（1.55±0.67）mmol/L、（4.83±0.53）mmol/L、（9.84±4.72）mU/L、（2.06±1.03）],差异均有统计学意义（P〈0.05或P〈0.01）;T2DM组HDL-C[（1.17±0.49）mmol/L]、HOMAβ-cell（55.81±25.84）低于对照组[（1.51±0.38）mmol/L、（76.79±51.56）],差异均有统计学意义（P〈0.05或P〈0.01）。②T2DM组血清SF[（5.84±0.69）ng/mL]、TRF[（13.32±5.88）ng/mL]高于对照组[（4.14±0.79）ng/mL、（9.14±2.93）ng/mL],差异有高度统计学意义（P〈0.01）。③2DM组血清SF水平分别与FINS、糖化血红蛋白（HbA1c）、HOMA-IR、BMI、WHR及SBP呈正相关,与HOMAβ-cell呈负相关（P〈0.01）;血清TRF水平分别与FINS、HOMA-IR、TG及WHR呈正相关（P〈0.01）。结论铁参与了T2DM发生、发展,SF、TRF可能是预测T2DM的因子。     

Risk factors of diabetic retinopathy in type 2 diabetic patients

Background Advances in treatment have greatly reduced the risk of blindness from this disease, but because diabetes is so common, diabetic retinopathy remains an important problem. The purpose of this study is to investigate the risk factors of diabetic retinopathy （DR） in Chinese type 2 diabetic patients.     

Proton magnetic resonance spectroscopy of normal human brain and glioma: a quantitive in vivo study

Invivoprotonmagneticresonancespectroscopy(MRS)providesanoninvasivemethodexaminingawidevarietyofcerebralmetabolitesin bothhealthysubjectsandpatientswithvariousbrain diseases.Thepossibilitythattumorshaveunique metabolicfingerprintsissupportedbyavarietyo studies.Inaqualitativecomparisonofinvivoproton MRSfindingsandhistologicaldatafromavarietyo braintumors,Bruhnetal1foundthathistologically similartumorsexhibitsimilarspectra,while histologicallydifferenttumorsexhibitquitedifferen spectralpatterns…     

UW液、Celsior液和HTK液低温保存生物人工肝用L-02细胞的效果比较

目的 比较UW液、Celsior液和HTK液4℃常规低温保存生物人工肝用L-02细胞的效果。方法 制备好的L-02细胞悬液分以下3组：UW液保存组（UW液组）;Celsior液保存组（CS液组）;HTK液保存组（HTK液组）。各组细胞于4℃低温保存72h后,分别测定细胞存活率及死亡率（流式细胞术）,ALT及LDH释放,尿素合成功能及白蛋白分泌功能。结果 UW液较Celsior液和HTK液显著提高了4℃常规低温保存保存72h的L-02细胞的存活率[(60.05 ± 4.23)% vs (50.12 ± 3.99)%、(44.20 ±4.67)%],均P<0.05;降低了细胞死亡率[(39.95±4.23)%vs (49.88 ±3.99)%、(55.80%±4.67)%], 均P<0.05;抑制了ALT、LDH释放(均P <0.05);更好地维持了L-02细胞尿素合成功能[ (1.03 ± 0.23)mmol/L vs (0.80± 0.14)mmol/L、(0.48± 0.05)mmol/L]和白蛋白分泌功能[(8.36 ±1.38 )mg/L vs (6.41±1.25)mg/L、(5.19±0.41)mg/L), 均P<0.05。结论 同Celsior液和HTK液相比,使用UW液4℃常规低温保存L-02肝细胞可以明显的提高复温后细胞存活率,降低低温损伤引起的ALT、LDH释放,有效的保护肝细胞尿素合成功能和白蛋白分泌功能,但保存时间不应超过72h。     

维持性血液透析患者透析前后血生化指标变化对自主神经功能的影响

目的探讨维持性血液透析(MHD)患者血液透析(HD)前后血生化指标变化对自主神经功能是否产生影响。方法选取2011年8月至2012年8月在广西壮族自治区人民医院进行治疗的80例MHD患者,HD前后测定心率变异性(HRV)时域指标及血生化指标。结果 MHD患者HD前后HRV时域指标的差异无统计学意义(P>0.05)。血钾(K+)、血尿素氮(BUN)、肌酐(Cr)HD前分别为(4.4±0.7)mmol/L、(21.6±7.5)mmol/L、(735.1±203.7)mmol/L,HD后分别为(3.6±0.5)mmol/L、(7.3±3.5)mmol/L、(271.0±73.2)mmol/L,HD后血K+、BUN、Cr均显著低于HD前(P<0.05)。血Ca2+、HCO3-HD前分别为(2.0±0.3)mmol/L、(21.9±2.5)mmol/L,HD后分别为(2.4±0.3)mmol/L、(27.6±2.1)mmol/L,HD后血Ca2+、HCO3-显著高于HD前(P<0.05)。各项生化指标与HRV各项指标间无显著相关性(P>0.05)。结论 MHD患者HD的过程不能改善心脏自主神经功能,HD前后血生化指标的快速变化对自主神经功能无明显影响。     

氨氯地平、瑞舒伐他汀联合非诺贝特治疗高血压合并冠心病的临床研究

目的 探讨氨氯地平、瑞舒伐他汀联合非诺贝特治疗高血压合并冠心病的临床疗效.方法 选择我院内科于2014年3月至2016年3月期间收治的98例高血压合并冠心病的患者为研究对象,根据随机数表法分为观察组和对照组,每组49例,对照组予氨氯地平及瑞舒伐他汀治疗,观察组在此基础上联合应用非诺贝特治疗,疗程3个月.比较两组患者治疗前后的血压、血脂及颈动脉内膜中层厚度(IMT)的变化.结果 治疗后,两组患者的收缩压、舒张压均较治疗前显著降低(P<0.05),但是组间比较差异均无统计学意义(P>0.05);观察组患者的甘油三酯(TG)、胆固醇(TC)、低密度蛋白-胆固醇(LDL-C)及颈动脉IMT分别为(1.84±0.57)mmol/L、(3.98±0.92)mmol/L、(2.25±0.68)mmol/L和(0.78±0.08)mm,均明显低于对照组的(2.21±0.79)mmol/L、(4.46±1.01)mmol/L、(3.19±0.79)mmol/L和(0.84±0.08)mm,高密度脂蛋白-胆固醇(HDL-C)为(1.39±0.21)mmol/L,明显高于对照组的(1.25±0.20)mmol/L,差异均有统计学意义(P<0.05).结论 氨氯地平、瑞舒伐他汀联合非诺贝特治疗高血压合并冠心病不仅可以降低患者血压,还可以有效调节血脂,延缓动脉粥样硬化进展.     

丙戊酸钠抑制肺癌细胞株A549增殖的研究

目的研究丙戊酸钠（sodium valproate,VPA）对肺癌细胞株A549细胞增殖的影响。方法体外不同浓度VPA不同作用时间处理人肺癌细胞株A549,倒置显微镜观察细胞形态变化,MTT比色法检测细胞增殖,流式细胞技术分析细胞周期与细胞凋亡的改变。结果2.0 mmol/L VPA作用72 h细胞形态明显改变、细胞数明显减少。1.0mmol/L VPA作用72 h后,细胞增殖被抑制,抑制率为（15.8±2.8）%,与同期对照组相比,差异具有统计学意义（P〈0.05）;提高处理浓度或延长作用时间,抑制作用有加强趋势。1.0 mmol/L VPA作用72 h后可改变细胞周期分布,G1期细胞比例为（72.2±3.4）%,与同期对照组相比差异显著;2.0 mmol/LVPA处理24～72 h后G1细胞比例由（64.5±3.7）%增加至（79.8±4.4）%,而S期细胞由（30.7±5.17）%降低至（14.2±3.37）%;提高处理浓度或延长作用时间均使G1期细胞比例增高,S期细胞比例减少。2 mmol/L VPA作用24～72 h可提高细胞凋亡率,由（7.30±1.87）%增加到（19.85±2.40）%。结论VPA可抑制肺癌A549细胞生长,改变细胞周期分布、促进细胞凋亡。     

儿童幽门螺杆菌感染与缺铁性贫血的相关性

目的 研究儿童幽门螺杆菌感染与缺铁性贫血的相关性.方法 选取我院2015年1月至2016年8月收治的120例儿童,依据胃镜及Hp检测结果将这些患儿分为Hp阳性慢性胃炎组(n=40)、Hp阳性胃镜下无损害组(n=40)和Hp阴性慢性胃炎组(n=40)三组,对三组患儿的缺铁性贫血指标、缺铁性贫血发生情况进行统计分析.结果 Hp阳性慢性胃炎组、Hp阳性胃镜下无损害组患儿的血红蛋白(Hb)、平均红细胞体积(MCV)、血清铁(SI)、血清铁蛋白(SF)水平[(113.3±11.4)g/L、(77.5±6.5)fl、(12.3±11.5)μmol/L、(16.5±8.2)μg/L和(116.3±13.2)g/L、(76.6±6.5)fl、(12.5±6.5)μmol/L、(20.3±12.5)μg/L]均显著低于Hp阴性慢性胃炎组[(133.5±7.5)g/L、(87.2±6.3)fl、(19.5±5.3)μmol/L、(35.2±11.5)μg/L](P<0.05),总铁结合力(TIBC)水平[(67.5±18.2)mmol/L、(73.5±18.2)mmol/L]均显著高于Hp阴性慢性胃炎组[(54.2±15.2)mmol/L],缺铁性贫血发生率[42.5%(17/40)、47.5%(19/40)]均显著高于Hp阴性慢性胃炎组[10.0%(4/40)],差异均有统计学意义(P<0.05),但Hp阳性慢性胃炎组、Hp阳性胃镜下无损害组患儿的Hb、MCV、SI、TIBC、SF水平、缺铁性贫血发生率比较差异均无统计学意义(P>0.05).结论 儿童幽门螺杆菌感染与缺铁性贫血密切相关,值得临床充分重视.     

青蒿琥酯增加人脑胶质瘤细胞CHG-5对β射线的敏感性

目的从青蒿素及其衍生物中筛选出对人脑胶质瘤细胞CHG-5及U87具有较好放射增敏效果的药物,并初步探讨其放射增敏机制。方法①采用MTT法测定上述青蒿素衍生物对2株胶质瘤细胞的IC50,观察以20%IC50浓度的青蒿素衍生物联合β射线对CHG-5及U87的放射增敏效果,从中筛选出放射增敏效果最好的药物;②通过克隆形成法计算目标药物的放射增敏比;③划痕法观察它对β射线照射后CHG-5细胞迁移的影响;④检测胞内还原型GSH水平;⑤流式细胞术检测细胞凋亡及细胞周期分布情况。结果青蒿素(artemisinin,ART)、双氢青蒿素(dihydroartemisinin,DHA)、青蒿琥酯(artesunate,AS)对CHG-5的IC50分别为:1.47、0.20、0.29mmol/L;对U87,则分别为1.61、0.25、0.46mmol/L。以20%IC50为β射线的协同给药浓度,仅有AS可以同时抑制经β射线照射后的CHG-5及U87细胞的增殖。且0.06mmol/L的AS对CHG-5细胞的放射增敏比为1.25,该浓度的AS可以明显地抑制CHG-5细胞的迁移。GSH结果表明0.06mmol/LAS处理组较未处理组细...     

单极与双极切割系统经尿道前列腺剜除术的比较

目的 探讨并比较单极与双极切割系统在经尿道前列腺解剖性剜除术(TUAEP)中的安全性和疗效。 方法 分别采用2种切割系统对良性前列腺增生(BPH)患者行TUAEP,单极98例,双极83例,比较2组的手术时间、前列腺切除质量、血红蛋白下降值、血Na⁺浓度、最大尿流率(Qmax)、残余尿(PVR)、国际前列腺症状评分(IPSS)、生活质量评分(QOL)等围手术期安全及疗效指标。 结果 采用2种切割系统均能将增生的前列腺腺体完整剜除,未发生输血、前列腺包膜穿孔和电切综合征等严重并发症。单极组及双极组的手术时间分别为(55.1±6.1)和(58.3±5.4)min,前列腺切除质量分别为(35.2±5.8)和(36.5±7.4)g,术后4 h血Na⁺浓度分别为(139.96±3.59)和(140.52±4.31)mmol/L,血红蛋白分别为(129.46±12.58)和(128.79±0.5)g/L,2组比较差别均无统计学意义(P>0.05)。术后6月,2组的前列腺特异抗原(PSA)、前列腺体积、Qmax、IPSS、QOL差别均无统计学意义(P>0.05),但与同组术前比较,PSA、前列腺体积、残余尿、Qmax、IPSS、QOL均较术前明显改善,差别有统计学意义(P<0.01)。 结论 采用单极或双极切割系统均可安全有效实施TUAEP,疗效满意,单极切割系统并不增加风险。     

替莫唑胺-尼莫地平-O6-苄基鸟嘌呤联用对胶质瘤细胞株U251抑制作用的研究

目的观察替莫唑胺（TMZ）、尼莫地平（NIM）、O6-苄基鸟嘌呤（O6-BG）联用对脑胶质瘤细胞株的抑制作用。探讨胶质瘤耐药发生的机制,寻找有效的多药耐药逆转剂,克服耐药性的产生,提高化疗效果,增加肢质瘤细胞对化疗药物的敏感性,最大限度地抑制胶质瘤细胞增殖,延缓肿瘤复发。方法用免疫细胞化学法检测胶质瘤细胞中MGMT蛋白表达,观察比较MGMT在用O6-BG处理前后表达的差异。分别以尼莫地平和O6-BG作为耐药逆转剂,与替莫唑胺单独或联合应用于体外培养的脑胶质瘤细胞U251。用MTT法检测细胞抑制率;观察TMZ、NIM、O6-BG联用对脑胶质瘤细胞的抑制作用。结果与对照组相比,MGMT在用O6-BG处理后表达明显减少。MTT比色法分别检测TMZ、TMZ＋NIM、TMZ＋O6-BG组及TMZ＋NIM＋O6-BG组对胶质瘤细胞U251的抑制作用,结果显示各组对胶质瘤细胞的抑制作用为TMZ＋NIM＋O6-BG联合组〉TMZ＋O6-BG〉TMZ＋NIM〉TMZ。与TMZ组比较,TMZ＋NIM组、TMZ＋O6-BG组对U251细胞株的抑制率显著升高,差异有统计学意义（P〈0．01）;同样,与TMZ组比较,TMZ＋NIM＋O6-BG组对U251细胞株的抑制率也显著升高,差异有统计学意义（P〈O．05）。结论O6-BG能够有效抑制MGMT表达,从而能逆转人脑胶质瘤细胞株U251对替莫唑胺的耐药性。尼莫地平、O6-BG二者均可增强替莫唑胺时脑胶质瘤细胞株U251的细胞毒作用,对于替莫唑胺的耐药有较好的逆转效果,因而可望作为临床化疗增敏剂与替莫唑胺联合使用。替莫唑胺、尼莫地平、O6-BG三者联用对肢质瘤细胞株U251有更好的抑制作用,其效果优于二者联合应用,强于单独应用替莫唑胺。     

丙戊酸钠在体内外影响NK细胞对人肺癌细胞杀伤作用的研究

目的观察丙戊酸钠(valproic acid,VPA)对人肺癌细胞A549、SP-CA-1、NCIH446中的MHCⅠ类链相关基因A(MICA)表达的影响,并比较经VPA处理前后NK细胞对肺癌细胞杀伤作用的差异,及肺癌荷瘤小鼠动物模型瘤组织重量、大小的差异。方法将0.75～12.0mmol/L VPA作用于A549、SP-CA-1、NCIH446细胞,与未加VPA的对照组进行比较,观察VPA对人肺癌细胞生长的影响。选择对细胞生长无影响的1.50mmol/L和3.00mmol/L VPA诱导三株肺癌细胞4d,通过RT-PCR反应检测MICA在mRNA水平的变化。LDH法检测VPA处理肺癌细胞前后其被NK细胞杀伤程度的变化。建立肺癌荷瘤小鼠动物模型,加入不同剂量VPA,体内观察VPA对肺癌瘤组织重量、大小的影响。结果用1.50mmol/L和3.00mmol/L VPA诱导4d后的三株肺癌细胞,与对照组比较,其MICA的mRNA转录水平提高,被NK杀伤的程度提高(P<0.05);高VPA浓度组三株肺癌细胞的mRNA水平及其被NK细胞杀伤的程度均高于低浓度组(P<0.05);VPA高剂量和/或低剂量组抑制荷瘤鼠皮下肿瘤的生长作用与对照组差异有统计学意义,P<0.05,抑瘤率分别为52.9%和35.9%。结论 VPA通过上调MICA抗肺癌作用的新机制,为肺癌临床免疫生物治疗提供了实验依据,具有治疗肺癌的潜在价值,VPA在体内对肺癌细胞A549的增殖也存在明显的抑制作用,为探讨肺癌治疗新方法及其机制提供一个新途径。     

RNA-binding Protein UNR Promotes Glioma Cell Migration and Regulates the Expression of Ribosomal Protein L9

Objective To investigate the role of RNA binding protein—upstream-of-N-Ras (UNR) in the development of glioma and its molecular mechanism.Methods First, bioinformatics analysis of CGGA database was performed to detect UNR expression level and prognosis of patients with glioma. Western blot and real-time PCR were used to detect UNR expression level in glioma cell lines and tissues. Next, UNR siRNAs were transfected in glioma cells, and MTS assay and scratch wound-healing assay were used to detect changes in cell proliferation and migration. Then, the candidate UNR target mRNAs were identified by analyzing the sequencing data of UNR iCLIP-seq, RNA sequencing and ribosome profiling databases of human melanoma. RNA immunoprecipitation and biotin pull-down assays were used to identify the UNR target mRNAs in glioma cells. Finally, western blot was used to detect the effect of UNR knockdown on ribosomal protein L9 (RPL9) and RPL9 protein expression level in glioma cell lines. RPL9 siRNA was transfected in A172 and T98G and the expression of vimentin in the cells was detected with western blot.Results Bioinformatics analysis showed that UNR mRNA expression level was significantly higher in high-grade glioma [Grade Ⅱ (n=126), Grade Ⅲ (n=51), Grade Ⅳ (n=128), P<0.001]. UNR high expression levels were associated with poor prognosis (P=0.0177). UNR had high expression level in glioma cell lines and patient samples compared with normal cell lines and normal brain samples (P<0.01). Knockdown of UNR inhibited glioma cells migration (P<0.05), but did not inhibit glioma cells growth in three glioma cell lines. UNR binded the 3’ untranslated region (UTR) of PTEN and RPL9 mRNAs. RPL9 protein was significantly highly expressed in most glioma cell lines (n=9) and knockdown of UNR resulted in a downregulation of RPL9 protein expression. Epithelial-mesenchymal transition (EMT)-related marker—vimentin was positively regulated by RPL9.Conclusions UNR could bind to the 3’UTR of PTEN and RPL9 in glioma cell lines, therefore promoting glioma cell migration and regulating the expression of RPL9. Here, we establish a link between UNR and RPL9 protein, which will provide new ideas for the further study of glioma.