MMP-2、TIMP-2与人胆管癌QBC939裸鼠移植瘤生长相关性的研究

目的探讨人胆管癌QBC939裸鼠移植瘤内基质金属蛋白酶-2(MMP-2)、金属蛋白酶组织抑制剂-2(TIMP-2)的表达与肿瘤发生、发展的关系。方法建立人胆管癌裸鼠移植瘤模型,并随机分成对照组、干预组(羟基喜树碱,2.5 g.kg-1)和实验组(羟基喜树碱,10 g.kg-1),观察裸鼠移植瘤体生长情况及瘤体病理组织学。结果所有移植瘤标本病理组织学检查符合胆管中分化腺癌;与对照组比,实验组可明显提高移植瘤内TIMP-2的含量,降低MMP-2/TIMP-2的比值;移植瘤体积与前者呈负相关,与MMP-2/TIMP-2呈正相关。结论实验组可降低移植瘤MMP-2/TIMP-2比值,抑制移植瘤的生长,对人胆管癌裸鼠移植瘤细胞外基质的降解可能具有抑制作用。     

热化疗对人胆管癌细胞增殖和凋亡的影响

目的探讨表阿霉素(EADM)热化疗对人胆管癌细胞(QBC939)的增殖和凋亡的作用。方法以体外培养的人胆管癌细胞株为研究对象,采用水浴加热法进行体外细胞毒实验(MTT)、透射电镜观察及流式细胞仪检测,观察热疗、化疗、热化疗对人胆管癌细胞的生长抑制和凋亡的影响。结果42℃以上单纯热疗对QBC939细胞有明显杀伤作用(P<0.01),42℃以上热化疗对QBC939细胞有明显的协同或相加作用(P<0.01)。透射电镜和流式细胞术均观察到热疗、化疗、热化疗诱导细胞凋亡的作用(P<0.01);热化疗有协同作用(P<0.01)。结论热疗、化疗、热化疗均抑制QBC939细胞增殖;热疗、化疗、热化疗诱导细胞凋亡为其抗癌机制之一;热疗提高化疗药物的细胞毒作用。     

RNA干扰降低环氧化酶2基因表达对人胆管癌QBC939细胞的体外作用

目的研究小干扰RNA（siRNA）降低环氧化酶2（COX-2）基因对胆管癌QBC939细胞凋亡及凋亡蛋白caspase-3蛋白表达的影响。方法采用RNA干扰的方法,将胆管癌细胞株QBC939细胞分为4组：COX-2 siRNA干预组、阴性对照siRNA组、空脂质体、空白对照组。将COX-2 siRNA转染入QBC939细胞;流式细胞仪检测siRNA降低COX-2基因表达对凋亡的作用,WesternBlot法检测siRNA降低COX-2基因表达对caspase-3表达变化的影响。结果 RNA干扰后QBC939细胞COX-2表达下降至空白对照组的42%,流式细胞仪检测结果显示COX-2 siRNA干预组细胞的凋亡率以及凋亡蛋白caspase-3的表达明显高于其他3个对照组。结论 RNA干扰可抑制细胞COX-2蛋白的表达,促使胆管癌QBC939细胞的凋亡,caspase-3途径可能是其调控通路之     

XPC Lys939Gln polymorphism,smoking and risk of sporadic colorectal cancer among Malaysians

AIM: To investigate the risk association of xeroderma pigmentosum group C (XPC ) Lys939Gln polymorphism alone and in combination with cigarette smoking on colorectal cancer (CRC) predisposition. METHODS: Peripheral blood samples of 510 study subjects (255 CRC patients, 255 controls)were collected. DNA was extracted and genotyping was performed using polymerase chain reaction-restriction fragment length polymorphism. The association between polymorphic genotype and CRC predisposition was determined using the OR and 95%CI. RESULTS: The frequency of the homozygous variant (Gln/Gln) genotype was significantly higher in cases compared with controls (16.0% vs 10.2%, P = 0.049). The Gln/Gln genotype of XPC showed a significantly higher association with the risk of CRC (OR = 1.884; 95%CI: 1.082-3.277; P = 0.025). In the case of allele frequencies, variant allele C was associated with a significantly increased risk of CRC (OR = 1.375; 95%CI: 1.050-1.802; P = 0.020). Moreover, the risk was markedly higher for those who were carriers of the Gln/Gln variant genotype and were also cigarette smokers (OR = 3.409; 95%CI: 1.061-10.949; P = 0.032). CONCLUSION: The XPC Gln/Gln genotype alone and in combination with smoking increases the risk of CRC among Malaysians.     

XAV939抑制人神经母细胞瘤细胞的增殖

目的探讨小分子抑制剂XAV939对人神经母细胞瘤(NB)细胞系SH-SY5Y细胞的抗增殖和诱导凋亡的作用,及其可能机制。方法采用MTT法检测XAV939对SH-SY5Y细胞活力的抑制作用,并确定最佳作用浓度。然后采用Annexin V-FITC法检测XAV939处理后早期凋亡细胞百分比,Hoechst33342染色法观察凋亡细胞核形态,克隆形成实验检测细胞体外增殖能力的变化。Western blotting检测Wnt/β-连环蛋白(β-catenin信号通路的关键蛋白和抗凋亡蛋白Bcl-2的变化。结果 MTT结果显示,1μmol/L XAV939作用24h后,SH-SY5Y细胞增殖的抑制率有明显上升。XAV939处理后48h及72h,凋亡细胞百分比显著高于对照组,且细胞呈现出不同程度的凋亡形态。此外,XAV939可显著降低SH-SY5Y细胞的体外克隆形成率,降低Wnt/β-catenin信号通路关键蛋白β-catenin,Cyclin D1和c-Myc及抗凋亡蛋白Bcl-2的表达。结论 XAV939可能部分通过抑制Wnt/β-catenin信号通路而抑制SH-SY5Y细胞的增殖。     

反义CD147分子对人胆管癌细胞株侵袭性影响的实验研究

目的探讨反义CD147分子对人胆管癌细胞株QBC939侵袭性的影响。方法构建含反义CD147分子片段的重组真核表达质粒,将人胆管癌细胞株QBC939分为三组:(1)反义CD147组,运用重组真核表达质粒as CD147-pc DNA3.1(-)转染QBC939细胞;(2)空质粒组,运用pc DNA3.1(-)空质粒转染QBC939细胞;(3)对照组,运用DMEM常规处理细胞。采用MTT法检测各组QBC939细胞的生长情况,RT-PCR和Western blotting法分别对三组QBC939细胞中的CD147、MMP-2、MMP-9、TIMP-2的m RNA表达水平及其对应蛋白的表达水平进行检测。结果成功构建了携带反义CD147分子片段的重组真核表达质粒。MTT法检测显示:三组QBC939细胞的生长曲线及对数生长期情况无统计学差异(P0.05)。RT-PCR法检测显示:反义CD147组QBC939细胞中CD147和MMP-2分子的m RNA表达均低于空质粒组和对照组(P0.05),且空质粒组与对照组相比无统计学差异(P0.05);反义CD147组QBC939细胞中MMP-9和TIMP-2分子的m RNA表达与空质粒组和对照组相比无统计学差异(P0.05)。Western blotting检测发现:与空质粒组和对照组相比,反义CD147组QBC939细胞中CD147和MMP-2分子的蛋白表达明显降低(P0.05),空质粒组与对照组相比无统计学差异(P0.05);与空质粒组和对照组相比,反义CD147组QBC939细胞株中MMP-9和TIMP-2分子的蛋白表达无统计学差异(P0.05)。结论 (1)反义CD147对胆管癌细胞株QBC939的生长无影响;(2)反义CD147对胆管癌细胞株QBC939中MMP-9和TIMP-2的m RNA表达及对应蛋白表达无影响;(3)反义CD147降低胆管癌细胞株QBC939中CD147和MMP-2的m RNA表达及对应蛋白表达,可能会降低胆管癌细胞株的侵袭性。     

Next generation histone deacetylase inhibitors: the answer to the search for optimized epigenetic therapies?

Introduction: HDAC inhibitors have demonstrated potent anticancer activities in preclinical and clinical studies. Currently, two drugs (SAHA and romidepsin) have gained the FDA approval for the treatment of cutaneous T-cell lymphoma. Clinical efficacy of HDAC inhibitors has been observed in advanced hematological malignancies, while response in other cancers has been in most cases unpredictable and often rather limited. The search for new molecules with the potential to overcome the limitations of the first HDAC inhibitors has become a primary goal in the field of epigenetic drug discovery as well as drugs acting on other chromatin modifying enzymes.

Areas covered: The article shortlists seven new HDAC inhibitors that have recently entered clinical studies as representative examples of next generation drugs. The most recently published preclinical profile is reviewed, together with the first clinical data for these compounds. The article then focuses on challenges faced during the progress of first generation HDAC inhibitors and analyzes whether these new compounds are likely to provide a solution to the existing issues and needs.

Expert opinion: Next generation HDAC inhibitors have the ‘best-in-class’ potential, particularly regarding potency and in vivo exposure. However, several issues remain unresolved. For example, none of the presented compounds appears to have a significantly different selectivity profile towards various HDAC isoforms and, thus, none of them may provide a further elucidation between the toxicity seen in more advanced HDAC inhibitors and isoform selectivity. Additionally, a need for a continuous effort on target validation is seen as a necessary requirement for further progress in the field.     

平消胶囊对胆管癌细胞生长及CD44v6蛋白表达的影响

目的通过观察平消胶囊(郁金、马钱子粉、仙鹤草、五灵脂、白矾、硝石、干漆、枳壳)血清作用于人肝外胆管癌细胞QBC939后,对癌细胞的生长和转移的影响,探讨其抗癌及转移机制。方法将平消胶囊灌喂大鼠,制备含药血清,血清培养胆管癌细胞,MTT法检测不同剂量含药血清对人胆管癌细胞QBC939的生长抑制作用,流式细胞仪检测细胞周期、细胞膜上黏附分子CD44v6表达的变化。结果平消胶囊血清能明显抑制人肝外胆管癌细胞QBC939的增殖,具有时间、剂量依赖关系。药物作用后,QBC939细胞生长受阻于G0/G1期,并且黏附分子CD44v6在细胞膜上表达下调,剂量越大,CD44v6下调越明显。结论平消胶囊能有效地抑制QBC939细胞增殖,通过影响细胞生长、诱导细胞膜上黏附分子CD44v6表达下调,导致细胞凋亡,防止癌细胞转移。     

p27~(kip1)真核表达载体的构建及其对人胆管癌细胞生长的影响

目的：构建p27~(kip1)真核表达载体并进行基因转染,研究其对人胆管癌细胞生长特性的影响。方法：将人p~(27kip1) cDNA克隆至真核表达载体pClneo的Not I酶切位点中,然后应用DOTAP脂质体将重组质粒转染人胆管癌QBC939细胞中,用G418筛选抗性细胞克隆,采用PCR、RT－PCR和Western印迹法检测外源性目的基因在靶细胞基因组中的整合及表达,以及转基因细胞对裸鼠致瘤性能力的改变。结果：成功构建p~(27kip1)基因真核表达载体,并将其导入QBC939细胞中。经G418筛选得到稳定表达p~(27kip1)基因的细胞株,p~(27kip1)基因转染细胞生长速度明显减慢,并出现分化的迹象,其裸鼠致瘤性能力明显降低。结论：p~(27kip1)基因具有抑癌基因的作用,在未来胆管癌的基因治疗中可能具有应用前景。