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1.
刘恒  王钢乐  李秀楠 《西部医学》2021,33(5):650-654
【摘要】目的 探讨microRNA-301a(miR-301a)对三阴性乳腺癌(TNBC)细胞的侵袭、增殖的调控作用及潜在的作用机制。方法 应用real-time PCR检测miR-301a在MDA-MB-231细胞与MCF-10A细胞中的表达;转染MDA-MB-231,按转染质粒分为Mimics miR-301a组、Mimics NC组、 Inhibitor miR-301a组、Inhibitor NC组,通过Transwell小室实验、MTT实验检测miR-301a对MDA-MB-231细胞侵袭及增殖能力的影响;应用生物信息学软件预测miR-301a的下游靶点,并应用荧光素酶报告实验以及Western blot分析miR-301a对靶点的调节作用。结果 miR-301a在MDA-MB-231细胞中的表达明显高于正常乳腺细胞(P<0.05);Transwell小室实验提示过表达miR-301a能够增强MDA-MB-231细胞的侵袭能力(P<0.05);MTT实验提示过表达miR-301a能够增强MDA-MB-231细胞的增殖能力(P<0.05);荧光素酶报告实验以及western blot实验证实miR-301a与MEOX2的靶向关系, Mimics miR-301a组MEOX2蛋白表达量明显低于Mimics NC组和空白组(P<0.05),Inhibitor miR-301a组MEOX2蛋白表达量明显高于Inhibitor NC组和空白组(P<0.05)。结论 miR-301a在TNBC中高表达,并可通过靶向作用于MEOX2促进TNBC细胞的侵袭、增殖,从而作为TNBC的促癌基因发挥作用。  相似文献   

2.
目的 研究miR-630在人体三阴性乳腺癌组织中的表达情况,探讨miR-630是否通过下调Sox4影响三阴性乳腺癌细胞系MDA-MB-231细胞的侵袭.方法 收集157例行乳腺癌根治术患者癌组织标本及癌旁正常乳腺组织,其中三阴性乳腺癌36例,通过实时荧光定量PCR(RT-PCR)检测各组(正常乳腺组、非三阴性乳腺癌组、三阴性乳腺癌组)组织中与乳腺癌相关的miR-630、miR-21、miR-195、miR-134、miR-200a、miR-381、miR-1228的表达情况;蛋白免疫印迹试验(Western blot)检测3组标本中细胞侵袭相关蛋白COL1 A1、COL1A5、MMP-2、MMP-9的表达情况;进一步在细胞实验中,通过转染miR-630mimic或空质粒后,再转染过表达Sox4的质粒或空质粒至离体培养的细胞中,Western blot检测细胞中COL1A1、COL1A5、MMP-2、MMP-9、Sox4的表达情况,划痕实验和Transwell法观察细胞迁移侵袭情况,利用荧光素酶实验验证Sox4是miR-630的靶基因.结果 相比于正常乳腺组,miR-630、miR-21在三阴性乳腺癌组织中的表达降低(P<0.01);miR-195、miR-134、miR-200a、miR-381、miR-1228的表达在3组间差异无统计学意义(P>0.05);相比于正常乳腺组及非三阴性乳腺癌组,三阴性乳腺癌组织中COL1A1、COL1A5、MMP-2、MMP-9、Sox4的表达增多(P<0.05).在细胞实验中,相比于空质粒组,转染miR-630 mimic后,MDA-MB-231细胞中COL1 A1、COL1A5、MMP-2、MMP-9、Sox4的表达减少(P<0.05),MDA-MB-231细胞的迁移侵袭能力减弱(P<0.01);荧光素酶实验结果显示,miR-630能够显著降低Sox4-3’-UTR质粒的荧光素活性(P<0.05);相比于过表达miR-630+空质粒组,转染过表达miR-630+过表达Sox4组,MDA-MB-231细胞中COL1A1、COL1A5、MMP-2和MMP-9表达增加(P<0.05),细胞的迁移侵袭能力增强(P<0.01).结论 在三阴性乳腺癌组织中,miR-630低表达;miR-630可以通过下调Sox4从而抑制MDA-MB-231的迁移和侵袭.  相似文献   

3.
目的 构建IRF-4结合蛋白(IBP)基因RNA干扰 (RNAi) 的真核表达载体,观察其对MDA-MB-231乳腺癌细胞内IBP基因表达及细胞增殖和侵袭力的影响.方法 以IBP为靶基因,以pcDNATM6.2-GW/EmGFPmiR质粒为载体,设计构建4对重组体,并进行DNA测序鉴定.重组载体转染MDA-MB-231细胞后,选择转染效果最好的1对重组体建立稳定转染的细胞株,经RT-PCR和Western blot检测重组表达质粒对IBP mRNA和蛋白表达的抑制效果;MTT法检测对各组细胞生长的影响;细胞体外侵袭实验测定对侵袭力的影响.结果 重组体测序结果与目的序列完全一致;重组体转染MDA-MB-231细胞后IBP的mRNA和蛋白表达降低约70%;IBP表达下调后乳腺癌细胞增殖减缓(P<0.05);同时体外侵袭实验显示转染后乳腺癌发生迁移细胞数明显低于未转染组和空质粒对照组[分别为(25±7)、(67±6)、(68±5),P<0.05].结论 下调IBP能有效降低乳腺癌细胞的增殖和侵袭力.  相似文献   

4.
目的 探究miR-378在乳腺癌细胞系中的表达及作用机制。方法 利用TargetScan分析miR-378的作用靶基因RAB11A。利用RT-PCR检测乳腺癌细胞系中miR-378和RAB11A的mRNA水平表达。转染MCF-7和MDA-MB-231细胞miR-378,NC(normal control)对照,采用MTT法和Transwell法检测乳腺癌细胞增殖和迁移能力的变化;采用RT-PCR和Western blot检测RAB11A的表达。荧光素酶报告分析miR-378与靶基因RAB11A之间的信号。结果 miR-378在乳腺癌细胞MCF-7,MDA-MB-231中表达上调;RAB11A在乳腺癌细胞MCF-7,MDA-MB-231中表达下调;在MCF-7,MDA-MB-231细胞中敲降miR-378后,乳腺癌细胞增殖和迁移能力与对照组相比被显著抑制,RAB11A的mRNA和蛋白表达水平明显增加,荧光素酶活性增加。结论 在乳腺癌细胞中,miR-378可通过靶向RAB11A促进乳腺癌细胞的增殖和迁移。  相似文献   

5.
谢铭芳  唐蒙蒙  丁伟 《浙江医学》2017,39(12):1005-1008
目的研究miR-630在人肺癌组织中的表达,探讨miR-630对肺腺癌细胞系(A549)细胞迁移侵袭的影响及其机制。方法收集37例肺腺癌组织标本及癌旁正常肺组织,通过RT-PCR检测各标本中miR-630、SOX4mRNA的表达情况,Pearson相关分析检验miR-630与SOX4的相关性;通过转染miR-630mimic和miR-630NC至离体培养的A549细胞中,Westernblot检测A549细胞中COL1A1、COL1A5、SOX4的表达情况,划痕试验和Transwell法观察细胞迁移侵袭情况,荧光素酶素试验验证SOX4是miR-630的靶基因。结果与癌旁正常肺组织比较,miR-630在肺腺癌组织中的表达降低(P<0.05);肺腺癌组织中SOX4mRNA的表达增加(P<0.01);miR-630与SOX4mRNA的表达量呈负相关(r=-0.344,P<0.01)。与对照组比较,转染miR-630mimic后A549细胞的迁移侵袭减少(P<0.01),细胞中COL1A1、COL1A5、SOX4的表达减少(均P<0.05);荧光素酶试验结果显示,miR-630能够降低SOX4-3′-UTR质粒的荧光素活性(P<0.05)。结论miR-630在肺腺癌组织中低表达;miR-630可能是通过下调SOX4抑制A549细胞的迁移和侵袭。  相似文献   

6.
目的探讨微小核糖核酸101(miRNA-101)在人乳腺癌MDA-MB-231细胞中的表达及其对MDAMB-231 细胞增殖的影响。方法实时荧光定量聚合酶链反应(qRT-PCR)检测乳腺癌细胞MDA-MB-231和正常乳腺上皮细胞MCF-10A 中miRNA-101 的表达。采用Lipofectamine TM 2000 将miRNA-101-mimic/inhibitor/NC 分别转染至MDA-MB-231 细胞中,通过qRT-PCR检测miRNA-101 的转染效率,CCK-8 实验检测MDA-MB-231 细胞的增殖。结果miRNA-101 在MDA-MB-231 细胞中的表达水平低于正常乳腺细胞MCF-10a( p<0.01)。转染miRNA-101 mimic 后MDA-MB-231 细胞的增殖能力减弱(p <0.05),而转染miRNA-101 inhibitor后细胞的增殖能力增强(p <0.05)。结论miRNA-101 在乳腺癌细胞MDA-MB-231中低表达,转染miRNA-101 mimic后乳腺癌细胞MDA-MB-231 的增殖减弱。  相似文献   

7.
目的 探究miR-21靶向E2F1对三阴性乳腺癌细胞恶性生物学活性及裸鼠肿瘤抑制率的影响。方法将MDA-MB-231细胞分为5组,即MDA-MB-231组、miR-21 inhibitor组、miR-NC inhibitor组、siRNA-E2F1组和siRNA-NC组。检测细胞中miR-21表达(RT-PCR法);分别检测细胞增殖(MTT法)、侵袭(Transwell法)、迁移(划痕实验)和凋亡能力(流式细胞仪);检测细胞中E2F1蛋白表达;检测miR-21与E2F1的靶向关系(双荧光素酶实验报告)。结果MDA-MB-231细胞中miR-21表达明显高于MCF10A细胞(P<0.05);miR-21 inhibitor组细胞细胞中miR-21表达明显低于MDA-MB-231组(P<0.05)。与MDA-MB-231组相比,miR-21 inhibitor组细胞吸光度值、侵袭能力、迁移能力和细胞中E2F1蛋白表达均明显降低,细胞凋亡能力明显升高(P<0.05);MDA-MB-231组细胞吸光度值、侵袭、迁移、凋亡能力和细胞中E2F1蛋白表达与miR-NC inhibitor组相比差异无统计学意义(P>0.05)。预测软件显示E2F1的3′UTR端与miR-21有碱基互补结合点位。通过向MDA-MB-231细胞中转染野生型E2F1(E2F1-WT)时,miR-21组荧光素酶活性明显低于miR-NC组(P<0.05);miR-21组和miR-NC组突变体荧光素酶活性相比差异无统计学意义(P>0.05)。与siRNA-NC组相比,siRNA-E2F1组细胞增殖、侵袭、迁移和细胞中E2F1蛋白表达均明显降低,细胞凋亡能力明显增加(P<0.05)。与miR-NC inhibitor组裸鼠移植肿瘤第8天时相比,miR-21 inhibitor组裸鼠肿瘤体积明显降低,肿瘤抑制率为45.3%(P<0.05)。结论低表达miR-21可抑制三阴性乳腺癌细胞增殖、侵袭,促进凋亡,且抑制裸鼠移植瘤体积,其作用机制可能与抑制E2F1表达有关。  相似文献   

8.
目的 利用三阴性人乳腺癌MDA-MB-231细胞,对稳定表达荧光素酶的细胞系MDA-MB-231-Luc进行功能评价及鉴定.方法 构建荧光素酶的HIV慢病毒载体,体外感染乳腺癌MDA-MB-231细胞,经嘌呤霉素筛选后得到稳定表达的细胞系,采用MTT法,利用Transwell侵袭模型,应用伤口愈合细胞划痕实验评价细胞增殖、侵袭和转移能力.结果 利用慢病毒转染的MDA-MB-231细胞能稳定表达荧光素酶,与亲本细胞相比,细胞的增殖、侵袭、迁移能力无显著影响(P>0.05).结论 慢病毒感染乳腺癌细胞能稳定表达荧光素酶,对MDA-MB-231细胞的增殖、侵袭、转移能力无明显影响,为后续的小动物活体成像实验提供细胞模型.  相似文献   

9.
目的探讨环状RNACdr1as/miR-7/SP1轴调控乳腺癌细胞发生侵袭和转移的作用机制。方法采用实时荧光定量PCR技术检测乳腺癌MDA-MB-231、MCF-7细胞转染miR-7模拟物、sh-Cdr1as、转录因子SP1siRNA前后,正常乳腺MCF-10A细胞及乳腺癌组织中环状RNACdr1as、miR-7、SP1mRNA的表达水平;MDA-MB-231、MCF-7细胞中E-钙黏蛋白(E-cadherin)及波形蛋白(Vimentin)mRNA的表达水平;采用Transwell实验分析MDA-MB-231、MCF-7细胞的侵袭和迁移能力;Westernblot法检测MDA-MB-231、MCF-7细胞中E-cadherin及Vimentin蛋白的表达情况。结果与MCF-10A细胞相比,MDA-MB-231、MCF-7细胞环状RNACdr1as表达上调,miR-7表达下调,SP1mRNA表达上调(均P<0.05)。与转染前相比,MCF-7、MDA-MB-231细胞转染sh-Cdr1as后,miR-7表达上调(均P<0.05),SP1mRNA表达下调(均P<0.05);而转染miR-7mimics后,SP1mRNA表达下调(均P<0.05)。与转染前相比,MDA-MB-231、MCF-7细胞转染sh-Cdr1as后,侵袭细胞数明显减少(均P<0.05)。与乳腺导管原位癌患者相比,乳腺浸润性导管癌环状RNACdr1as表达上调(P<0.05);与无腋窝淋巴结转移患者相比,有腋窝淋巴结转移的乳腺癌患者环状RNACdr1as表达上调(P<0.05);Pearson相关性分析显示环状RNACdr1as与miR-7的表达存在负相关(r=-0.541,P<0.05),miR-7与SP1mRNA的表达也存在负相关(r=-0.467,P<0.05)。MDA-MB-231、MCF-7细胞转染SP1siRNA后,E-cadherinmRNA、蛋白表达均上调(均P<0.05),VimentinmRNA、蛋白表达均下调(均P<0.05)。结论乳腺癌细胞环状RNACdr1as表达上调,通过环状RNACdr1as/miR-7/SP1调控轴促使癌细胞发生上皮间质转化,使乳腺癌细胞发生侵袭和转移。环状RNACdr1as可能成为乳腺癌治疗的新靶点。  相似文献   

10.
《中国现代医生》2020,58(36):32-35+封三
目的 探讨多能蛋白聚糖(VCAN)通过PI3K/AKT 信号通路对乳腺癌细胞MDA-MB-231 增殖的影响。方法 利用Western Blot 和qRT-PCR 检测VCAN 在正常人乳腺细胞MCF-10A 和三株乳腺癌细胞中的表达水平。选取VCAN 高表达细胞株MDA-MB-231 为实验对象,实验分组为NC 组、siRNA1-VCAN 组和siRNA2-VCAN 组。采用MTT 法及克隆形成法检测转染后的乳腺癌细胞增殖变化。Western Blot 检测p-AKT、AKT、p-PI3K、PI3K 的蛋白相对表达水平。结果 与正常人乳腺细胞相比,VCAN 在乳腺癌细胞中的表达量显著上升(P<0.05)。下调VCAN 表达量,MDA-MB-231 细胞的增殖显著被抑制(P<0.05)。同时,下调VCAN 表达量,p-AKT、p-PI3K 蛋白水平显著下降(P<0.05)。结论VCAN 在乳腺癌MDA-MB-231 细胞中,可能通过PI3K/AKT 信号通路调控细胞增殖能力。  相似文献   

11.
FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…  相似文献   

12.
Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P < 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (>0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P <0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P < 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.  相似文献   

13.
Objective: To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods: SD rats served as both donors and recipients. 4℃ sodium lactate Ringer's was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava (IVC) inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results: Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion: This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.  相似文献   

14.
Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P< 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P< 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.  相似文献   

15.
Objective: To observe the therapeutic effects in acupunture treatment of primary dysmenorrhea combined with spinal Tui Na, and study its mechanism. Methods: Thirty cases of the treatment group were treated by acupuncture combined with spinal Tui Na, and thirty cases in the control group were treated by routine acupuncture. Results: The total effective rate was 93.3% in the treatment group, and 73.3% in the control group, with a significant difference between the two groups (P<0.05). Conclusions: Acupuncture combined with spinal Tui Na has good prospects for treatment of primary dysmenorrhea.  相似文献   

16.
In treating chronic nephropathy,Luo Lingjie,a chief physician,pays attention to regulating the balance between yin and yang,treating infection if present,and removing pathogenic factors.He prescribes gentle drugs and uses carefully strongly warming-tonifying ones,emphasizes the importance of persuading the patient to persist in treatment with medication and nurse one's health for recuperation,and is good at combined use of TCM and western medicine therapy and brings the merits of various therapies into full play,with obvious theraoeutic effects.  相似文献   

17.
Dr.Zhang Ren,the chief physician,is the chairman of Shanghai Acupuncture and Moxibustion Association.Having been engaged in medicine for about 40 years,he is experienced in treating various intractable diseases.In his long years of clinical practice,he advocates taking the TCM differentiation as the basis to seek for the acupuncture method for treatment of modern intractable diseases.The author of this essay had the fortune to follow Dr.Zhang in study.The following is a summary of Dr.Zhang's experience in the acupuncture treatment for different intractable diseases with the same therapeutic principle.  相似文献   

18.
目的:评价使用安心颗粒对急诊经皮冠状动脉介入术(PPCI)术后生活质量的影响.方法:将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组(术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次)和对照组(仅接受基础药物治疗).所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表(MIDAS)、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果:入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异(P>0.05);出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低(P<0.05);组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低(P<0.05).两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血(P>0.05).两组发生轻度血小板减少症的患者数比较无差异(P>0.05).结论:PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.  相似文献   

19.
20.
目的 探讨猪肺磷脂注射液联合经鼻持续气道正压通气(NCPAP)对呼吸衰竭早产儿的临床疗效及肌酸激酶同工酶活性(CK-MB)的影响.方法 选取呼吸衰竭早产儿80例,分为观察组和对照组各40例.对照组采用NCPAP给氧治疗,观察组给予NCPAP给氧联合猪肺磷脂气管内给药.观察两组患儿治疗前及治疗12h、24 h后PaO2、PaCO2、血氧饱和度(SaO2)、pH的变化情况,检测治疗前及治疗5d后血清CK-MB水平;评估两组患儿的临床治疗效果.结果 两组患儿PaO2、PaCO2、SaO2、pH比较,差异均有统计学意义(P<0.05),其中观察组治疗后的PaO2、SaO2、pH均高于对照组,PaCO2则低于对照组.两组的PaO2、SaO2、pH均随观察时间延长而升高(P<0.05),PaCO2均随观察时间的延长而降低(P<0.05).观察组治疗有效率为87.5%,显著高于对照组的70.0% (P <0.05).治疗5d后两组患儿血清CK-MB水平均较前降低(P<0.05),且观察组明显低于对照组(P<0.05).结论 猪肺磷脂注射液气管内给药联合NCPAP可以显著降低呼吸衰竭早产儿CK-MB的含量,提高治疗有效率,起到很好的呼吸循环支持作用.  相似文献   

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