MIF在幽门螺杆菌L型相关胃癌中的表达及临床意义

目的研究巨噬细胞移动抑制因子(MIF)在胃癌组织中的表达及临床意义,并探讨其机制。方法应用免疫组化(SP法)和革兰染色检测80例胃癌组和50例对照组的Hp-L型感染,免疫组化(SP法)检测上述组织MIF蛋白表达。结果胃癌组MIF表达阳性率高于其对照组(P<0.05),且表达水平与浸润深度、淋巴结转移、临床分期、分化程度有关(P<0.05),与性别、年龄无关(P>0.05);革兰染色、免疫组化两种方法检测Hp-L型的结果具有一致性(P<0.05),胃癌组与对照组的Hp-L型阳性率相比具有统计学意义(71.25%vs 14.00%,P<0.05)。胃癌组Hp-L型感染阳性率仅与淋巴结转移有关(P<0.05),Hp-L型阳性组的MIF表达阳性率高于Hp-L型阴性组(P<0.05),且Hp-L型阳性和MIF阳性呈正相关(r=0.598,P<0.05)。结论 MIF蛋白表达与胃癌的浸润转移密切相关,其机制可能与幽门螺杆菌L型(Hp-L型)感染有关。     

TLR2在胶质瘤细胞GL261的高表达及其激活后增强迁移的作用

目的探讨小鼠胶质瘤细胞系GL261主要高表达的Toll样受体（TLRs）家族成员及其在GL261细胞迁移中的作用。方法运用实时荧光定量PCR检测TLRs在GL261细胞的表达情况;GL261细胞经TLR2配体Pam3CSK4刺激后,应用Transwell迁移实验检测其对迁移能力的影响;慢病毒稳定干扰TLR2表达后,观察Pam3CSK4刺激对GL261细胞迁移能力的改变。结果GL261细胞主要高表达TLRs家族中的TLR2;Pam3CSK4激活TLR2后显著增强GL261细胞迁移能力;沉默TLR2基因表达后,Pam3CSK4促进GL261细胞迁移能力的作用消失。结论小鼠GL261胶质瘤细胞TLRs家族中主要高表达TLR2,其激活具有促进GL261细胞迁移的作用,下调TLR2可抑制该细胞的迁移。     

一氧化氮介导的脂联素抑制HSC-T6的迁移作用研究

目的探讨脂联素对肝星状细胞-T6细胞系（hepatic stellate cell-T6 cell line,HSC-T6）细胞迁移作用机制,并评价一氧化氮（n itric oxide,NO）在此过程中的作用。方法以脂联素治疗肝HSC-T6细胞,采用qPCR检测肝脏星状细胞的iNOS基因表达水平;W estern b lot法检测iNOS蛋白的表达以及划痕修复实验检测脂联素作用下HSC-T6的迁移情况。结果脂联素作用下,iNOS基因及蛋白表达均明显升高,HSC-T6细胞培养基中NO代谢产物浓度显著升高,HSC-T6细胞迁移显著受抑制。结论脂联素具有抑制HSC-T6细胞迁移的作用,这种作用可能是通过NO介导实现的。     

Glutathione-S-transferase enhances proliferation-migration and protects against shikonin-induced cell death in breast cancer cells

Glutathione-S-transferase (GST) is a cytoplasmic protein responsible for detoxification, but the effect of the enzyme on cell biological events, including proliferation and migration, has never been reported. Thus, we evaluated the detoxification effect of in vitro–applied GST on cancer cell proliferation and migration. Assays for proliferation and migration of human breast cancer cells in the presence of GST were carried out. Binding of GST on the surface of the cancer cells was studied by flow cytometry. Detoxification through GST pathway was studied in the presence of shikonin. The effective dosage of GST in enhancement of cell proliferation was 10–50 nM, and the cell migration could be significantly enhanced after 6 hours in the presence of 2–50 nM GST. Therefore, overall cell proliferation and migration could be enhanced in the presence of 10 nM or greater concentration of GST, and 15 μM shikonin-induced toxification of the cancer cells could be neutralized by 1.0 μM GST. Flow cytometry showed that GST directly bound to the surface of the cancer cells, and this was confirmed by fluorescence confocal microscopic observation. It is concluded that human class π-GST enhances proliferation and migration of human breast cancer cells by means of direct binding to the cell surface and maintaining cell viability by detoxification.     

Nuanced but significant: How ethanol perturbs avian cranial neural crest cell actin cytoskeleton,migration and proliferation

Children with fetal alcohol syndrome (FAS) display striking craniofacial abnormalities. These features are proposed to result from perturbations in the morphology and function of cranial neural crest cells (cNCCs), which contribute significantly to the craniofacial complex. While certain pathways by which this may occur have been suggested, precise teratogenic mechanisms remain intensely investigated, as does the question of the teratogenic dose. The present study focused on examining how avian cNCC actin cytoskeleton, migratory distance, and proliferation are affected ex vivo by exposure to ethanol concentrations that simulate maternal intoxication. Chick cNCCs were cultured in 0.2% and 0.4% v/v ethanol. Distances migrated by both ethanol-treated and control cells at 24 and 48 h were recorded. Following phalloidin immunocytochemistry, treated and control cNCCs were compared morphologically and quantitatively. Apoptosis and proliferation in control versus treated cNCCs were also studied. Chick cNCCs cultured in ethanol lost their spindle-like shapes and their ordered cytoskeleton. There was a significant stage-dependent effect on cNCC migration at 24 h (p = 0.035), which was greatest at stage 10 (HH). Ethanol treatment for 48 h revealed a significant main effect for ethanol, chiefly at the 0.4% level. There was also an interaction effect between ethanol dose and stage of development (stage 9 HH). Actin microfilament disruption was quantitatively increased by ethanol at the doses studied while cNCC proliferation was increased but not significantly. Ethanol had no effect on cNCC apoptosis. At ethanol levels likely to induce human FAS, avian cNCCs exhibit various subtle, potentially significant changes in morphology, migration, and proliferation, with possible consequences for fated structures.     

中高度海拔地区世居与移居汉族慢性胃炎患者胃黏膜内MDA、SOD、XOD、NO含量比较

目的：探讨高原地区世居与移居汉族人群慢性胃炎胃组织内四种体液因子代谢水平差异及其意义.方法：检测高海拔地区（2 000m～2 500m）（西宁市及周边县区）世居与移居汉族人群慢性胃炎胃组织内氧自由基、脂质过氧化物丙二醛（MDA）、超氧化物歧化酶（SOD）、一氧化氮（NO）、黄嘌呤氧化酶（XOD）水平.结果：相同海拔高度地区世居与移居汉族人群胃黏膜内MDA、SOD、NO与XOD含量或活力无明显差异.结论：平原地区人群移居高原后同样发生了严重的氧自由基与氧自由基清除剂失衡,而且高原习服后的上述因子的水平与同海拔高度地区的世居汉族相同.     

壮药扶正方对TAMs与肿瘤细胞共培养体系A549细胞生物学行为的影响

目的:探讨壮药扶正方含药血清对肿瘤相关巨噬细胞（TAMs）与肺癌A549细胞共培养体系肺癌A549细胞增殖、迁移及侵袭等生物学行为影响及可能的作用机制。方法:采用氟波酯（PMA）联合白细胞介素-4、白细胞介素-13诱导人急性白血病单核细胞株（THP-1）建立TAMs体外模型,实时定量PCR及酶联免疫吸附试验法分析含药血清（低、中、高剂量组)及药物处理后24、48、72 h对TAMs表型调节;采用共培养体系,以肺癌A549细胞为研究对象,分别采用MTT法、实时细胞分析技术（RTCA）、Transwell法检测不同条件下A549细胞增殖、迁移及侵袭情况。结果:壮药扶正方含药血清能显著上调TMAs中M1型标志物白细胞介素-12、诱导型一氧化氮合酶表达水平,下调M2型标志物白细胞介素-10、CD206表达水平;与空白对照组比较,共培养组A549细胞增殖明显增加,细胞迁移及侵袭能力均明显升高,差异有统计学意义（P<0.05）;与共培养组比较,壮药扶正方含药血清低、中、高剂量组A549细胞增殖、迁移和侵袭作用受抑制,且呈剂量依赖性（P<0.05）。结论:壮药扶正方含药血清在共培养体系中能够抑制肺癌A549细胞的增殖、迁移及侵袭作用,机制可能与其调节肿瘤微环境TAMs表型有关。     

长链非编码RNA LOC101927476抑制卵巢癌细胞侵袭迁移和增殖

目的探讨长链非编码RNA(LncRNA)LOC101927476在卵巢癌细胞中的表达及其对卵巢癌生物学特征的影响。方法选取2018—2019年于中国医学科学院肿瘤医院手术治疗的卵巢癌患者。采用实时荧光定量PCR(real-time PCR)检测卵巢癌细胞系3AO、OVCA429、TOV21G、A2780、SKOV3以及22例卵巢癌原发瘤和转移瘤组织中LncRNA LOC101927476的表达水平, 采用TCGA数据库中卵巢癌转录组测序数据验证LncRNA LOC101927476和GATA4的表达, 采用慢病毒包装体系构建过表达LOC101927476(OE-LOC101927476组)和空载(OE-EV组)慢病毒并转染至3AO细胞。设计单链向导RNA(sgRNA), 利用CRISPR/Cas9构建LncRNA LOC101927476敲除细胞系。采用Transwell法和划痕实验检测LncRNA LOC101927476对卵巢癌细胞侵袭和转移能力的影响。采用细胞计数盒8(CCK-8)法检测细胞的增殖能力。结果 real-time PCR显示, 22例卵巢癌患者中, 20例转移灶中L...