细胞外囊泡装载仑伐替尼对肝癌细胞株HepG2增殖、侵袭及凋亡的影响

目的 探讨细胞外囊泡装载仑伐替尼对肝癌细胞株HepG2增殖、侵袭及凋亡的影响。方法 设肝癌细胞株HepG2组、细胞外囊泡组(extracellular vesicles, EVs混悬液5 mL、106个/mL)、仑伐替尼组(仑伐替尼5 mL、30 mg/L)、仑伐替尼载药囊泡组(EVs混悬液5 mL、106个/mL+仑伐替尼5 mL、30 mg/L),以上各组细胞每孔设6个平行样,培养72 h。培养结束后,测定细胞增殖、侵袭、迁移、凋亡水平,应用RT-PCR法和蛋白印迹法测定各组细胞miR-482、CYR61表达水平。结果 与肝癌细胞株HepG2组比较,细胞外囊泡组OD值、存活率、单克隆形成数目、穿膜数、迁移距离、凋亡率、miR-482、CYR61 mRNA和蛋白水平差异无统计学意义(P>0.05)。与肝癌细胞株HepG2组和细胞外囊泡组比较,仑伐替尼组、仑伐替尼载药囊泡组OD值、存活率、单克隆形成数目、穿膜数、迁移距离、CYR61 mRNA和蛋白水平降低,凋亡率、miR-482升高(P<0.05)。与仑伐替尼组比较,仑伐替尼载药囊泡组OD值、存活率、单克隆形成数目、穿膜数、迁移距离、CYR61 mRNA和蛋白水平降低,凋亡率、miR-482升高(P<0.05)。结论 细胞外囊泡装载仑伐替尼后能明显增强仑伐替尼对肝癌细胞株HepG2的杀伤力,增加其抑制增殖、侵袭与促凋亡作用;其机制可能与细胞外囊泡装载仑伐替尼能促进肝癌细胞株HepG2高表达miR-482,低表达CYR61有关。

FCRL5蛋白对肝癌细胞增殖与侵袭迁移能力的作用及机制

目的探讨FCRL5蛋白对肝癌细胞增殖与侵袭迁移能力的作用及机制。方法测定104例肝癌组织及癌旁组织FCRL5蛋白的表达水平,同时将SMMC-7721肝癌细胞株分为对照组(SMMC-7721细胞常规培养)和FCRL5转染组(FCRL5-mimics转染SMMC-7721细胞、FCRL5-inhibitor转染SMMC-7721细胞),MTT法检测肝癌SMMC-7721细胞活力,RT-PCR法测定FCRL5 mRNA表达水平,酶联免疫吸附试验(ELISA)测定FCRL5、基质金属蛋白酶(MMP)-9、G1/S-特异性周期蛋白(cyclin)D1、血管内皮生长因子(VEGF)蛋白表达水平,Transwell细胞迁移实验测定SMMC-7721细胞侵袭迁移能力。结果肝癌组FCRL5蛋白表达水平、半定量水平显著高于癌旁组(P<0.05);肝癌组FCRL5蛋白阳性率显著高于癌旁组(P<0.05)。FCRL5转染组、存活率、单克隆形成数目显著高于对照组,凋亡率显著低于对照组(P<0.05);FCRL5转染组FCRL mRNA、FCRL蛋白表达显著高于对照组(P<0.05);FCRL5转染组的穿膜数、侵袭距离显著高于对照组(P<0.05);FCRL5转染组MMP-9、cyclinD1、VEGF表达显著高于对照组(P<0.05)。FCRL5 inhibitor组单克隆形成数目、穿膜数、FCRL5 mRNA、FCRL5蛋白、MMP-9、cyclinD1、VEGF表达水平显著低于对照组、FCRL5转染组(P<0.05),凋亡率显著高于对照组、FCRL5转染组(P<0.05)。结论FCRL5在肝癌组织中高度表达;FCRL5具有促进肝癌细胞增殖与侵袭迁移能力的作用,其机制与FCRL5可能诱导肝癌细胞高表达MMP-9、cyclinD1、VEGF有关。

复方苦参注射液联合阿帕替尼治疗非小细胞肺癌的疗效观察

目的探讨复方苦参注射液联合阿帕替尼治疗非小细胞肺癌患者的临床疗效。方法选取2016年1月至2020年1月间重庆市合川区人民医院收治的102例非小细胞肺癌患者,采用随机抽签法分为观察组和对照组,每组51例。对照组患者采用阿帕替尼治疗,观察组患者采用复方苦参注射液联合阿帕替尼治疗,比较两组患者治疗效果、不良反应及肿瘤标志物。结果观察组患者近期治疗有效率为35.3%,对照组为17.6%,两组比较,差异有统计学意义(P <0.05)。观察组患者骨髓抑制、血红蛋白减少和肝肾功能损伤不良反应率均低于对照组,差异均有统计学意义(均P <0.05)。治疗前,两组患者癌胚抗原(CEA)、糖蛋白抗原125 (CA125)和细胞角单白-19片段(CYFRA21-1)比较,差异无统计学意义(P> 0.05)。治疗后,观察组患者CEA、CA125和CYFRA21-1均低于对照组患者,差异均有统计学意义(均P <0.05)。结论非小细胞肺癌患者采用复方苦参注射液联合阿帕替尼治疗,可获得良好的疗效,确保瘤体缩小或消失,且减轻药物不良反应,值得推广。

Chemokine SR-PSOX/CXCL16 expression in peripheral blood of patients with acute coronary syndrome

Background Scavenger receptor that binds phosphatidylserine and oxidized lipoprotein/CXC chemokine ligand 16 （SR-PSOX/CXCL16） promotes foam cell formation through the tumor necrosis factor （TNF）-α mediated mechanism. Because chemokine CXCL16 could be expressed in atherosclerotic lesions and induce smooth muscle cell （SMC） proliferation, we＇presume that the monocyte SR-PSOX/CXCL16 detection in the patients＇ peripheral blood will be important for early diagnosis and prognosis of atherosclerosis （AS）. Methods Enrolled in this study were 40 patients with acute coronary syndrome （ACS）, including 20 patients with acute myocardial infarction （AMI） and 20 patients with unstable angina pectoris （UAP）, and 20 normal controls. Monocytes in the peripheral blood were isolated, and the changes of expression of CXCL16/SR-PSOX mRNA were compared using reverse transcription-polymerase chain reaction （RT-PCR）, with β-actin as internal control. We compared the expression of CXCL16/SR-PSOX in the ACS subgroups, using Western-blot to analyze protein expression levels. Tissue sections were made from biopsy specimens taken from patients with infective endocarditis, liver cirrhosis, and lung cancer as well as normal controls. And the expression of CXCL16/SR- PSOX was analyzed with a confocal microscope. Results The expression of CXCL16/SR-PSOX mRNA and protein in the monocytes of peripheral blood was significantly higher in ACS patients than in normal controls （P〈0.05）; however, there was no significant difference in CXCL16/SR-PSOX expression between UAP group and AMI group （P〉0.05）. Immunofluorescence showed that there were low expression of SR-PSOX in normal vascular endothelial cells and enhanced expression in every layer of the infected vessels, while spreading from endothelial cells to surrounding tissues as infection worsens. Confocal microscopy showed that the expression of SR-PSOX was enhanced in the infiltrated lymphocytes in liver cirrhosis, and that the expression level was proportionate to the degree of inflammation in the portal hepatis and folia. Conclusions The expression of CXCL16/SR-PSOX in the monocytes of peripheral blood was significantly higher in ACS patients than in the controls. CXCL16/SR-PSOX-mediated inflammation may contribute to the pathogenesis of ACS, and CXCL16 may play an important role in the pathogenesis and development of AS in humans.

Fc受体样4蛋白在肝癌组织中的表达及对癌细胞Huh7生物学行为的影响

目的探讨Fc受体样4蛋白(FCRL4)在肝癌组织中的表达及对癌细胞Huh7生物学行为的影响。方法选取本院肿瘤科冰冻肝癌肿瘤组织及其邻近癌旁正常组织各40份,免疫组化法测定癌旁组、肝癌组中FCRL4的表达水平。设Huh7细胞组、FCRL4mimics组、FCRL4inhibitor组,培养72h后,测定细胞活力、凋亡、侵袭水平,RT-PCR法及蛋白印迹法测定各组细胞FCRL4基因和蛋白水平。结果肝癌组FCRL4蛋白阳性率明显高于癌旁组(P<0.05)。FCRL4mimics组OD值、存活率、穿膜数、FCRL4 mRNA和蛋白水平高于Huh7细胞组,FCRL4inhibitor组OD值、存活率、穿膜数、FCRL4mRNA和蛋白水平低于Huh7细胞组和FCRL4inhibitor组(P<0.05)。FCRL4mimics组凋亡率低于Huh7细胞组,FCRL4inhibitor组凋亡率高于Huh7细胞组和FCRL4inhibitor组(P<0.05)。结论肝癌中FCRL4过表达,FCRL4过表达能促进Huh7癌细胞增殖、侵袭,抑制其凋亡。