Inhibitory Effects of Parthenolide on the Angiogenesis Induced by Human Multiple Myeloma Cells and the Mechanism

The inhibitory effects of parthenolide （PTL） on angiogenesis induced by multiple myeloma （MM） cells in vitro and the mechanism were investigated. Human MM line RPMI8226 cells were cultured in vitro. The effects of MM culture supernatant on the migration and tubule formation ability of human umbilical vein endothelial cells （HUVECs） treated with PTL were observed. By using Western blot, the expression of p65 and IкB-α in MM cells was detected. RT-PCR was used to assay the expression of VEGF, IL-6, MMP2 and MMP9 mRNA in MM cells. ELISA was used to measure the levels of VEGF and IL-6 in MM cell culture supernatant. The expression of MMP2 and MMP9 in MM cells was examined by immunohistochemistry. （1） In 3.5, 5.0, 7.5 and 10 μmol/L PTL groups the number of migrated cells was 310±56, 207±28, 127±21 and 49±10 respectively, which was significantly different from that in positive control group （598±47） （P〈0.01）. In 3.5 and 5.0 μmol/L PTL groups the areas of capillary-like structures were 0.092±0.003 and 0.063±0.002 mm2, significantly less than in positive control group （0.262±0.012 mm2） （P〈0.01）, but in 7.5 and 10 μmol/L PTL groups no capillary-like structures were found; （2） After treatment with different concentrations of PTL for 48 h, the expression of p65 protein was gradually decreased, while that of IкB-α was gradually enhanced with the increased concentration of PTL; （3） After treatment with 3.5, 5.0, 7.5 and 10 μmol/L PTL for 48 h, the VEGF levels in the supernatant were 2373.4±392.2, 1982.3±293.3, 1247.0±338.4 and 936.5±168.5 pg/mL respectively, significantly different from those in positive control group （2729±440.0 pg/mL） （P〈0.05）. After treatment with 7.5 and 10 μmol/L PTL, the IL-6 levels in the culture supernatant were 59.6±2.8 and 41.4±9.8 pg/mL respectively, signifi- cantly lower than in positive control group （1287.3±43.5 pg/mL） （P〈0.05）; （4） RT-PCR revealed that PTL could significantly inhibit the expression of V     

不同代次骨髓间充质干细胞成骨分化潜能的体外研究

目的观察不同代次人骨髓间充质干细胞(MSC)体外成骨分化能力,为优化组织工程种子细胞提供更多参数。方法应用全骨髓直接贴壁法分离培养10例健康志愿者MSC,依照年龄将MSC分为4组,<20岁,20岁～,30～40岁,>40岁组。取P1(passage 1)～P5诱导成骨分化,诱导分化14d予以碱性磷酸酶染色,21d茜素红染色,Image-Pro Plus软件分析细胞染色阳性区域平均吸光度值。结果 <20岁组P1～P5成骨分化能力无显著差异(P>0.05);30～40岁组P3分化能力低于P1、P2(P<0.05);>40岁组P3分化能力下降更明显(P<0.05or P<0.01),P4、P5几乎分化受阻。结论不同代次的MSC分化能力具有异质性,与年龄有关。>40岁组MSC在超过P3代时,细胞倍增时间明显延长,诱导成功率降低,成骨分化潜能明显下降,选择P2代作为种子细胞更有利于细胞治疗的安全及质量控制。     

EGCG对多发性骨髓瘤细胞KM3促血管新生作用的抑制效应及机制初探

本研究探讨绿茶茶多酚成分之一表没食子儿茶素没食子酸酯[(-)-epigallocatechin-3-gallate,EGCG)]对多发性骨髓瘤细胞KM3促血管新生作用的影响及其机制。体外培养多发性骨髓瘤细胞KM3,观察不同浓度EGCG作用后的培养上清对血管内皮细胞株HUVEC迁移和形成血管能力的影响。ELISA法检测KM3细胞培养上清中血管内皮生长因子(VEGF)分泌水平;RT-PCR方法检测KM3细胞VEGF mRNA表达水平。结果表明:EGCG以浓度依赖方式抑制KM3细胞培养上清促内皮细胞迁移的作用,5,25,50和100μmol/L的EGCG迁移细胞数目分别为414±27,299±70,202±42及116±13个,且EGCG的浓度与内皮细胞的迁移数目呈负相关(r=-0.952,p<0.05)。同时,内皮细胞形成小管的面积随EGCG浓度的升高而减少,25及50μmol/L时的面积分别是88343.9±3231.1和60897.5±914.1μm2,均明显低于对照组(p<0.01),小管面积与EGCG浓度呈负相关(r=-0.888,p<0.05)。5、25、50、100μmol/L的EGCG作用于KM3细胞48小时后培养上清中VEGF的含量分别为1399.0±47.4pg/ml,660.1±5.7pg/ml,108.5±5.8pg/ml和26.2±18.6pg/ml,与对照组比较,25、50、100μmol/L组均有统计学意义(p<0.01)。RT-PCR结果显示EGCG抑制KM3细胞VEGF的mRNA水平表达,且呈浓度依赖性。结论:EGCG能显著抑制多发性骨髓瘤细胞KM3的促血管新生作用;下调VEGF转录水平的表达,减少VEGF的分泌可能是其药理作用机制之一。     

微泡：肿瘤微环境的信息载体

肿瘤微环境由肿瘤细胞自身及周围的基质细胞组成。肿瘤细胞与微环境的信息交流在肿瘤的发生及进展中起着“帮凶”作用。细胞间直接接触及可溶性细胞因子为其经典的对话方式,而细胞活化后释放的微泡,一种不同于二者的庞大信息载体,选择性组装传递核酸、蛋白等,提供肿瘤与微环境对话的新平台。本文就微泡在肿瘤细胞与肿瘤微环境相互对话中的生物...     

多发性骨髓瘤患者血清OPG检测及临床意义

目的：探讨OPG在多发性骨髓瘤（MM）患者血清中的表达水平及与骨病、预后的关系,初步探索其在MM骨病和预后判断中的作用。方法：采用双抗体夹心酶联免疫吸附试验（ELISA）技术分别检测28例MM患者和28例对照组的血清中OPG的水平。结果：MM患者的血清OPG水平为（222．4&#177;114．8）pg／ml,与对照组相比明显降低（P〈0．01）。0～1个骨损组血清OPG水平为（312．4&#177;129．6）pg／ml,而2～3个骨损组为（179．0&#177;59．4）pg／ml,0～1个骨损组明显高于2～3个骨损组（P〈0．01）。MM患者血清OPG水平与β2-微球蛋白相关（P〈0．01）,与球蛋白相关（P〈0．01）,与白蛋白无关（P〉0．05）,与临床分期、免疫学分型、血红蛋白、血钙未发现有相关性。结论：MM患者血清OPG水平明显低于正常对照,且其降低程度与骨损害的严重程度密切相关;同时,OPG水平与β2-微球蛋白呈负相关,提示血清OPG水平可能对MM的预后具有重要意义。     

Enhanced Response of Acute Monocytic Leukemia Cells to Low-dose Cytarabine by 1,25-dihydroxyvitamin D3

Low-dose cytarabine combined with differentiating or DNA hypomethylating agents, such as vitamin D compounds, is a potential regimen to treat acute myeloid leukemia (AML) patients who are unfit for high-intensity chemotherapy. The present study aimed to determine which subset of AML would be most responsive to low-dose cytarabine with the differentiating agent 1,25-dihydroxyvitamin D3 (1,25-D3). Here, firstly, cBioPortal database was used and we found out that vitamin D receptor (VDR) was highly expressed in acute monocytic leukemia (M5) and high VDR expression was associated with a poor survival of AML patients. Then, we confirmed that 1,25-D3 at clinical available concentration could induce more significant differentiation in acute monocytic leukemia cell lines (U937, MOLM-13, THP-1) and blasts from M5 patients than in non-monocytic cell lines (KGla and K562) and blasts from M2 patient. Finally, it was shown that the combination of 1,25-D3 and low-dose cytarabine further increased the differentiating rate, growth inhibition and G0/G1 arrest, while mild changes were found in the apoptosis in acute monocytic leukemia cell lines. Our study demonstrates that the enhanced response of acute monocytic leukemia cells to low-dose cytarabine by 1,25-D3 might indicate a novel therapeutic direction for patients with acute monocytic leukemia, especially for elderly and frail ones.     

认识意义未明特发性血细胞减少症

意义未明特发性血细胞减少症概念的提出,是为了便于定义临床上排除了其他血液系统和非血液系统疾病、且不符合骨髓增生异常综合征(MDS)诊断标准的一类单系或多系血细胞减少性疾病,有必要正确认识其疾病特点.本文对ICUS的诊断、鉴别及处理原则等方面作一述评.     

氨磷汀在体外减轻足叶乙甙所致的骨髓间充质干细胞损伤

目的研究氨磷汀在体外对化疗药物所致的骨髓间充质干细胞（MSCs）损伤的影响，为其应用于慢性粒细胞白血病（CML）患者的造血干细胞移植提供实验依据。方法采集CML患者的骨髓细胞，应用极限稀释法获取单克隆来源的MSCs，并在低血清培养液中培养和扩增。按实验分组分别加入氨磷汀（氨磷汀组）、足叶乙甙（VP-16组）、氨磷汀和足叶乙甙（联合处理组），并设没有处理的对照组。在药物作用一定时间后，计数MSCs，绘制生长曲线；流式细胞仪检测MSCs的免疫表型及凋亡率；在甲基纤维素半固体培养中检测MSCs作为滋养层支持造血的能力。结果MSCs生长曲线显示，与对照组相比，氨磷汀组的MSCs生长未受明显影响，联合处理组MSCs的生长显著好于沿16组。培养48h后，对照组、氨磷汀组、联合处理组和VP-16组的MSCs凋亡率分别为（4．9±0．8）％、（5．6±1．2）％、（18．7±3．4）％和（33．8±5．1）％，氨磷汀组与对照组相比，差异无统计学意义；联合处理组高于对照组，但明显低于VP-16组（P〈0．05）。形成粒-单核系集落、红系集落和多系集落的产率，氨磷汀组与对照组间的差异无统计学意义，联合处理组低于对照组（P〈0．05），但显著高于VP-16组（P〈0．05）。氨磷汀作用前后未见MSCs免疫表型发生明显改变。结论氨磷汀在体外对CML患者骨髓来源MSCs的增殖、凋亡、造血支持能力以及免疫表型无明显影响，但能显著改善足叶乙甙对MSCs所致的生长抑制及功能损伤，提示在预处理中加用氨磷汀可能成为促进CML患者造血干细胞移植后造血恢复的一种新策略。     

候选抑癌基因RIZ1

视网膜母细胞瘤蛋白质结合的锌指蛋白1( RIZ1)是一种甲基转移酶,其功能单位为特征性的PR锌指结构域.RIZ1能够通过PR结构域甲基化组蛋白3上第9位的赖氨酸,导致相应基因转录受抑,具有抑制肿瘤发生的作用.RIZ1在多种人类肿瘤中表达减低或缺失,其表达水平减低与肿瘤发生、进展等密切相关,被定义为一种候选抑癌基因.RIZ1在肿瘤中表达减低机制既涉及细胞遗传学又与表观遗传学改变密切相关.