高浓度葡萄糖诱导人晶状体上皮细胞发生EMT

目的:观察高浓度葡萄糖诱导人晶状体上皮细胞发生上皮-间质转分化(epithelial-to-mesenchymal transition,EMT)。方法:将人晶状体上皮细胞HLE-B3系分别培养在正常葡萄糖浓度(5.5 mmol/L)DMEM培养基和高浓度葡萄糖(35.5 mmol/L)的DMEM培养基中24小时,于培养的0 h、3 h、6 h、12 h、24 h在倒置显微镜下观察细胞形态学变化,采用免疫荧光染色检测晶状体上皮细胞中EMT相关蛋白E-cadherin及α-SMA的表达变化。结果:与正常糖浓度组相比,随着时间的延长高糖组细胞逐渐丢失上皮细胞形态,细胞变细、变长,向纤维细胞的形态转变;同时随着时间的延长,高糖组晶状体上皮细胞中E-cadherin染色的荧光强度在各时间点均低于正常糖浓度组,而α-SMA的荧光强度却明显高于正常糖浓度组,在6 h和12 h时差异显著,有统计学意义(P0.01)。结论:高浓度葡萄糖诱导人晶状体上皮细胞发生上皮-间质转分化。     

STZ诱导的糖尿病模型小鼠肾脏的上皮-间质转分化研究

目的:探讨STZ诱导的糖尿病小鼠肾脏发生上皮-间质转分化(EMT)的情况。方法:将80只C57BL小鼠随机分为正常对照组(NC组)和糖尿病组(DM组),每组40只。DM组小鼠用1%STZ(streptozotocin,链脲佐菌素)溶液按60mg/kg体质量的剂量进行腹腔注射,每天1次,连续6天。NC组小鼠平行腹腔注射同等体积0.1mol/L的柠檬酸钠缓冲液。再将成模小鼠随机分为A、B批次,A批次用于动态观察生存率、小鼠体质量及随机血糖的监测;B批次用于在造模后第4、8、12周末观察肾组织的病理变化,并用Western blot、免疫荧光染色的方法观察肾组织中EMT标志蛋白α-SMA和E-cadherin的表达。结果:STZ诱导的糖尿病模型小鼠出现糖尿病典型症状如多饮、多尿等,血糖持续在高水平状态,体质量增长缓慢。在造模后12周末,DM组小鼠较NC组小鼠累积生存率显著降低,两组比较差异具有统计学意义(P0.001)。在造模后的第8周末,DM组小鼠肾脏出现明显的病理改变,到第12周末时,绝大部分肾小管上皮细胞被梭形的肌成纤维细胞取代,肾小球空泡,基底膜增厚。造模后的第4、8、12周末时,DM组小鼠E-cadherin表达量均显著低于NC组小鼠(P=0.004,0.026,0.004);而在第8和12周末时,DM组α-SMA表达量显著升高(P=0.009,0.015)。在第12周末,肾组织冰冻切片E-cadherin和α-SMA、免疫荧光染色结果与上述结果一致。结论:STZ诱导的糖尿病模型小鼠有较典型的糖尿病临床改变,且肾组织发生了EMT。     

雌激素对A549细胞系EMT过程的影响

目的:探讨雌激素对A549细胞系EMT标志物表达量的影响。方法:用不同浓度的雌激素刺激A549细胞系,并用q-RT-PCR和Western-blot实验检测各组细胞中EMT标志物表达量的变化,用Transwell实验检测不同浓度雌激素对细胞迁移能力的影响,计算各组间有无统计学差异。结果:当雌激素浓度为1×10-9 mol/L、1×10-8 mol/L、1×10-7 mol/L时,Vimentin的m RNA表达量分别为:2.14±0.55、4.72±0.63、2.21±0.47,显著高于空白对照组,组间有统计学差异,E-cadherin的m RNA表达量分别为:0.64±0.15、0.46±0.11、0.59±0.13,显著低于空白对照组,组间有统计学差异,蛋白表达量也有同样趋势;细胞迁移数分别为58.63±7.33、80.12±9.32、61.89±8.22,组间有统计学差异。当雌激素浓度为1×10-8 mol/L时,Vimentin的表达量最高,E-cadherin的表达量最低,细胞迁移数最高。结论:适宜浓度雌激素可以促进Vimentin的表达,抑制E-cadherin的表达,提高细胞迁移能力,当雌激素浓度为1×10-8 mol/L时,促进Vimentin表达、抑制E-cadherin表达和促进细胞迁移的作用最显著。由此认为,雌激素对A549细胞系发生EMT过程有促进作用。     

刺参糖胺聚糖抗仙台病毒作用机制的探讨

目的:探讨刺参糖胺聚糖的抗仙台病毒(Sendaivirus,SV)作用及其作用机制.方法:选用不同浓度刺参糖胺聚糖作用于仙台病毒感染BHK21细胞的多个环节,倒置显微镜观察病毒致细胞痛变效应,MTT法检测细胞活性;同时以SV滴鼻感染小鼠,观察刺参糖胺聚糖对感染病毒小鼠血凝抑制抗体产生的影响.结果:刺参糖胺聚糖浓度大于3.2mg/ml时表现出细胞毒性作用.浓度在0.25～0.2mg/ml范围时,刺参糖胺聚糖作用病毒吸附组和病毒复制组能明显抑制细胞病变,MTT结果也表明该两组显示较好的细胞活性,且具有一定的量效关系,无细胞毒性作用;但SJ-GAG直接作用病毒组、预处理细胞组以及抑制病毒穿入组无明显抗病毒作用.动物实验表明刺参糖胺聚糖可促进小鼠血凝抑制抗体的产生.结论:刺参糖胺聚糖的抗仙台病毒作用主要通过抑制病毒对靶细胞的吸附以及抑制病毒复制而实现,并能促进小鼠对病毒感染的免疫应答.     

异氟烷对小鼠星型胶质细胞Sirt1和MAO-A基因表达的影响

目的:探讨异氟烷对小鼠星形胶质细胞Sirt1和MAO-A基因表达的影响。方法:给予体外培养的新生小鼠原代星形胶质细胞不同浓度异氟烷处理,实验分为对照组和异氟烷处理组(0.5ISO、1.0ISO、1.5ISO),其中异氟烷组细胞分别给予0.5 MAC、1.0MAC和1.5 MAC三个浓度的异氟烷处理2小时,对照组给予O_2处理2小时,然后提取细胞RNA和蛋白检测Sirt1和MAO-A的mRNA和蛋白表达变化。结果:与对照组相比,异氟烷下调小鼠星形胶质细胞的Sirt1和MAO-A mRNA和蛋白水平,异氟烷浓度越大下调越明显。结论:异氟烷可抑制小鼠星形胶质细胞的Sirt1和MAO-A基因表达。     

高表达Snail蛋白诱导黑色素瘤EMT细胞模型的构建及鉴定

目的:构建稳定表达Snail蛋白的可用于肿瘤上皮-间质化研究的肿瘤细胞模型.方法:采用亚克隆方法从质粒pCMV6-mSnail中PCR扩增小鼠Snail基因,连接至表达质粒pL-tdTomato-Neo,筛选重组质粒并经双酶切及测序鉴定.构建成功的重组质粒pL-tdTomato-mSnail转染小鼠黑色素瘤B16细胞,G418筛选稳定细胞株.采用荧光定量PCR和Western blot技术检测胞内Snail及上皮/间质标志物的变化.建立裸小鼠皮下移植瘤模型,活体成像系统观测移植瘤.结果:重组质粒pL-tdTomato-mSnail成功构建,其稳定转染株B16/dT-mSN胞体发出强烈红色荧光.胞内Snaill水平显著上调,E-钙粘蛋白下调,波形蛋白表达上调,呈现典型的上皮-间质化表型.结论:成功获得稳定高表达小鼠Snail蛋白的EMT细胞模型,且可用于体内外荧光成像观测,为研究Snail蛋白在介导肿瘤EMT过程中的生物作用提供了重要的实验工具.     

c-myb对小鼠体外受精的影响及作用机制的分析

目的: 本文采用免疫组织化学方法观察Myb转录因子家族成员c-myb蛋白在小鼠精子和卵母细胞-卵丘复合物中的分布.方法: 建立小鼠体外受精模型,用不同浓度反义c-myb寡脱氧核苷酸(c-myb ASODNs)与精子、卵母-卵丘细胞复合物共孵育观察其对小鼠体外受精率的影响,并进一步探讨c-myb ASODNs对小鼠体外受精率的影响机制.结果: 在颗粒细胞胞核和精子的头部有c-myb蛋白分布.c-myb ASODNs呈剂量依赖性抑制小鼠体外受精率(小鼠体外受精率在空白组、低、中、高浓度c-myb ASODNs组、无义tat ODNs组分别为34.97%、30.89%、20.14%、16.68%、34.47% ).GABA、P4、Verapamil和dbcAMP与c-myb ASODNs共同作用时,GABA、P4、dbcAMP三者均逆转c-myb ASODNs对受精的抑制作用,(小鼠体外受精率在空白组、中浓度c-myb ASODNs组、GABA、P4、Verapamil和dbcAMP组、分别为34.81%、22.96%、40.83%、39.12%、7.463% 、40.61%),GABA、P4、和dbcAMP三者对精子中的c-myb蛋白免疫组化染色阳性的细胞百分率无明显影响.Verapamil能抑制小鼠体外受精,并协同c-myb ASODNs的抑制作用,且使精子中的c-myb蛋白免疫组化染色阳性的细胞百分率明显下降.结论: c-myb ASODNs与受精密切相关, Verapamila可能通过调节精子中的c-myb表达,从而抑制小鼠体外受精,而GABA、P4和dbcAMP可能不是通过c-myb来影响受精过程.     

高浓度葡萄糖对小鼠囊胚Caspase-8表达的影响

目的:探讨高浓度葡萄糖对小鼠囊胚Caspase-8表达的影响.方法:通过促超排卵,获取妊娠3.5d小鼠囊胚,随机分成三组,即对照组(空白)、低糖组(葡萄糖浓度为7.5mmol/L)和高糖组(葡萄糖浓度为28.0mmol/L),分别培养在含0、7.5mmol/L和28.0mmol/L葡萄糖的M199培养基中,培养24h后,然后再吸出囊胚.每组随机吸取30个囊胚用免疫组织化学S-P法.检测不同浓度葡萄糖对小鼠囊胚Caspase-8表达状况,利用HPIAS-1000图像分析系统测定Caspase-8在以上三组中表达的平均光密度和平均阳性面积率.结果:Caspase-8表达结果:空白组中囊胚细胞胞浆中可见少量浅棕黄色颗粒,Caspase-8表达呈弱阳性.低糖组中囊胚细胞胞浆未见着色,Caspase-8表达呈阴性.高糖组囊胚细胞胞浆中可见较多的棕黄色颗粒,Caspase-8表达呈强阳性.空白组与低糖组囊胚Caspase-8表达的阳性面积率及平均光密度无显著性差异(P>0.05),高糖组与空白组和低糖组相比均存在显著性差异(P<0.01).结论:高浓度葡萄糖可诱导Caspase-8的过度表达,导致囊胚细胞数目过度减少,从而影响囊胚的正常发育和着床.     

子宫内膜异位症患者免疫调节Th17细胞及Treg细胞的表达意义

目的探究子宫内膜异位症患者免疫调节Th17细胞及Treg细胞的表达意义。 方法选取2017年1月至2018年12月青岛大学附属医院收治患有子宫内膜异位症的患者,为子宫内膜异位组（EMT组）,选取同一时期在医院因不孕不育进行腹腔镜检查的患者,为正常组（NM组）,两组分别56例。EMT组和NM组患者在一般资料上差异无统计学意义。通过流式细胞仪、HE染色法、qRT-PCR法、ELISA法分析EMT组和NM组患者Th17、Treg细胞所占比例、子宫内膜组织病变情况、ROR-γt、Foxp3 mRNA表达含量的差异性来探究子宫内膜异位症患者Th17细胞及Treg细胞变化。实验结果用 ±s表示,并采用独立样本t检验进行比较。 结果EMT组患者CD4⁺ T细胞中Th17所占比例为5.48±2.81,Treg所占比例为4.22±1.04,NM组Th17所占比例为2.34±1.01,Treg所占比例为6.14±1.52,差异均有统计学意义（t = 7.869,3.015,P = 0.014,0.026）。EMT组患者血清中IL-17水平为（256.38±34.15）?pg/ ml、IL-22为（67.48±10.89）?pg/ml,NM组患者血清中IL-17水平为（198.04±27.59）?pg/ml、IL-22为（43.78±6.92）?pg/ml,差异均有统计学意义（t = 9.944,4.689,P = 0.008,0.017）。EMT组患者血清中IL-10水平为（18.56±4.77）?pg/ml、TGF-β为（148.28±40.52） pg/ ml,NM组患者血清中IL-10水平为（28.35±6.07）pg/ml、TGF-β为（204.78±19.87）pg/ml,差异均有统计学意义（t = 9.491,2.849,P = 0.012,0.034）。EMT组患者子宫内膜组织形态不规则,多数细胞不完整,破损或缺失,且炎性细胞增多,在其周围聚集。NM组患者子宫内膜组织形态规则,细胞没有明显破损或缺失,未见细胞周围炎性因子增多。qRT-PCR检测结果显示,EMT组和NM组ROR-γ mRNA分别为2.89±0.76、1.71±0.26,EMT组和NM组Foxp3 mRNA分别为2.25±0.34、1.13±0.18,两组差异均有统计学意义（t = 10.996,6.759,P = 0.006,0.011）。 结论子宫内膜异位症患者外周血免疫调节细胞Th17/Treg平衡失调,免疫调节紊乱与子宫内膜异位发生、发展有密切关系。     

Tbx18通过下调EMT信号分子参与调控心脏发育

转录因子Tbx18(Tbx18)在小鼠胚胎心外膜上皮细胞表达并调控心外膜上皮细胞向心系细胞分化.上皮间充质转化(EMT)过程是器官发育和形成的重要机制.为阐述Tbx18通过调控下游EMT关键信号分子参与心外膜上皮细胞分化和心脏发育,本研究运用Tbx18-Cre/Rosa26R-EYFP双杂合基因敲入小鼠和免疫荧光共聚焦,证实Tbx18+心系细胞和EMT关键信号分子Snail1、Smad、Slug、Twist在发育后期胚鼠心外膜和心外膜下间充质发生共聚焦.同时还发现,Tbx18在胚鼠不同发育阶段的表达模式和Tbx18+心系细胞内上述EMT关键信号分子的表达模式相似.Tbx18和EMT关键信号分子在发育心脏存在相似的时空表达模式,因此,它们之间可能存在相互调控作用.运用Tbx18突变技术揭示了Tbx18突变型胚鼠心脏EMT关键信号分子表达水平均较野生型显著下调,直接证实了上述4个EMT信号分子是Tbx18的可能靶点.理解Tbx18参与心脏发育的下游靶点有助于改善成年心脏损伤后的再生修复.     

肿瘤相关成纤维细胞对非小细胞肺癌恶性生物学行为影响

目的:探讨肿瘤相关成纤维细胞(Tancer Associated Fibroblast,TAF)对非小细胞肺癌(Non-small Cell Lung Cancer,NSCLC)恶性生物学行为的影响。方法:选取在本院肿瘤科住院手术的非小细胞肺癌患者,收集术后肺癌标本,马松三色染色(Masson Trichrome Stain)和天狼星红染色(Sirius Red Stain)观察肺癌组织(Lung Cancer Tissue,LCT)、癌旁组织(Pericarcinomatous Tissue,PCT)和正常组织(Normal Tissue,NT)中TAF的表达情况;体外将非小细胞肺癌细胞A549与非小细胞肺癌成纤维细胞P-gp共培养,CCK-8检测共培养前后A549细胞增殖能力;细胞划痕和Trans-well实验分别检测A549细胞迁移和侵袭能力;q RT-PCR和Western blot检测A549细胞上皮间质转化(Epithelial Mesenchymal Transition,EMT)标志蛋白E-cadherin、N-cadherin和Vimentin的表达。结果:Masson和Sirius染色结果显示:肺癌组织中纤维的表达明显高于癌旁组织;与P-gp共培养的A549细胞的增殖、迁移和侵袭能力及上皮间质转化相关蛋白N-cadherin和Vimentin表达均明显高于阴性对照组(P0.05),而E-cadherin的表达明显降低(P0.05)。结论:TAF可能通过诱导非小细胞肺癌细胞EMT的发生从而促进非小细胞肺癌的增殖、迁移和侵袭等恶性生物学行为。     

环状GMP-AMP合成酶高表达对乳腺癌细胞上皮间质转化的影响

目的:探讨环状GMP-AMP合成酶(cGAS)高表达对乳腺癌MCF7细胞发生上皮间质转化(EMT)的影响。方法:构建稳定高表达cGAS的慢病毒载体并转染MCF7细胞;转染后细胞分别培养12 h,24 h,48 h,72 h,每组实验重复三次,采用MTT检测cGAS对MCF7细胞增殖的影响; transwell法检测高表达cGAS对MCF7细胞迁移能力的影响;蛋白免疫印迹(Western blot)法分析EMT相关蛋白E-cadherin和N-cadherin的表达情况。结果:与对照组比较,cGAS上调后MCF7细胞增殖能力显著增强(P<0.05);细胞形态学观察显示cGAS上调后诱导MCF7细胞EMT发生,细胞形态由鹅卵石样变为梭形; transwell实验结果显示,cGAS上调导致MCF7细胞迁移能力增强(P<0.05); Western blot结果表明,cGAS上调后上皮标记蛋白E-cadherin表达下降(P<0.05),间质标记蛋白N-cadherin表达增加(P<0.05)。结论:cGAS上调可增强乳腺癌细胞的增殖和迁移能力,诱导乳腺癌细胞EMT发生...     

miR-125a-3p调控人结肠癌细胞浸润转移的作用及机制研究

目的:探讨miR-125a-3p在结肠癌细胞浸润与转移中的作用及其可能机制。方法:通过qRT-PCR方法检测miR-125a-3p在结肠癌细胞及组织样本中的表达;在结肠癌细胞过表达或沉默miR-125a-3p后,通过平板克隆实验、MTT实验、划痕实验、Transwell实验检测结肠癌细胞增殖、迁移及侵袭能力的变化;采用Western blot方法检测miR-125a-3p过表达后相关标志分子的表达水平变化情况。结果:miR-125a-3p在结肠癌细胞及组织呈现异常低表达;过表达miR-125a-3p抑制结肠癌细胞HCT116及SW480的增殖能力;过表达或沉默miR-125a-3p分别抑制或增强结肠癌细胞的迁移与侵袭能力;过表达miR-125a-3p在mRNA及蛋白水平均能够显著抑制Snail、N-cadherin及Vimentin的表达,而增加E-cadherin的表达。结论:miR-125a-3p参与调节结肠癌细胞浸润与转移,其机制可能是通过调控上皮间质转化途径介导的。     

Fractionated radiotherapy might induce epithelial-mesenchymal transition and radioresistance in a cellular context manner

Despite the fact that radiotherapy is a main therapeutic modality in cancer treatment, recent evidence suggests that fractionated radiotherapy (FR) might confer radioresistance through epithelial-mesenchymal transition (EMT). Nevertheless, the effects of FR on EMT phenotype and the potential link between EMT induction and radioresistance development yet to be clarified. The aim of this study was to assess whether FR could promote EMT, and to elucidate if induction of EMT contributes to the acquisition of radioresistance. To this end, two human cancer cell lines (A549 and HT-29) were irradiated (2 Gy/day) and analyzed using wound healing, transwell migration and invasion assays, real-time polymerase chain reaction (for E-cadherin, N-cadherin, Vimentin, CD44, CD133, Snail, and Twist), clonogenic assay, Annexin V/PI, and 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay. Irradiation of A549 (for 5 or 10 consecutive days) resulted in morphological changes including elongation of cytoplasm and nuclei and pleomorphic nuclei. Also, irradiation-enhanced migratory and invasive potential of A549. These phenotypic changes were in agreement with decreased expression of the epithelial marker (E-cadherin), enhanced expression of mesenchymal markers (N-cadherin, Vimentin, Snail, and Twist) and increased stemness factors (CD44 and CD133). Moreover, induction of EMT phenotype was accompanied with enhanced radioresistance and proliferation of irradiated A549. However, FR (for 5 consecutive days) did not increase HT-29 motility. Furthermore, molecular alterations did not resemble EMT phenotype (downregulation of E-cadherin, Vimentin, ALDH, CD44, CD133, and Snail). Eventually, FR led to enhanced radiosensitivity and decreased proliferation of HT-29. Altogether, our findings suggest that FR might induce EMT and confer radioresistance in a cell context-dependent manner.     

GABAB receptor inhibits tumor progression and epithelial-mesenchymal transition via the regulation of Hippo/YAP1 pathway in colorectal cancer

Gamma-Aminobutyric Acid Type B Receptor (GABABR) plays essential roles in tumor progression. However, the function of GABABR in colorectal cancer (CRC) needs further clarification. As the main part of GABABR, GABABR1 expression was identified significantly lower in tumor tissues than those in non-tumor normal tissues and that CRC patients with high GABABR1 expression lived longer. Further studies indicated that knockdown of GABABR1 elevated CRC cell proliferation, migration, and invasion. Furthermore, knockdown of GABABR1 activated the expression of the epithelial-mesenchymal transition (EMT)-related proteins N-cadherin and Vimentin, whereas decrease the protein level of E-cadherin. In addition, activation of Hippo/YAP1 signaling contributes to the GABABR1 down-regulation promoted proliferation, migration, invasion and EMT in CRC cells. At last, we verified the contribution of Hippo/YAP1 signaling in the GABABR1 down-regulation impaired biological phenotype of colon cancer cells in vivo. In summary, these data indicate that GABABR1 impairs the migration and invasion of CRC cells by inhibiting EMT and the Hippo/YAP1 pathway, suggesting that GABABR1 could be a potential therapeutic target for CRC.     

Overexpression of heat shock protein 70 inhibits epithelial-mesenchymal transition and cell migration induced by transforming growth factor-β in A549 cells

Heat shock protein 70 (HSP70) is a key member of the HSP family that contributes to a pre-cancerous environment; however, its role in lung cancer remains poorly understood. The present study used geranylgeranylacetone (GGA) to induce HSP70 expression, and transforming growth factor-β (TGF-β) was used to construct an epithelial-mesenchymal transition (EMT) model by stimulating A549 cells in vitro. Western Blot was performed to detect protein levels of NADPH oxidase 4 (NOX4) and the EMT-associated proteins E-cadherin and vimentin both before and after HSP70 expression. Cell morphological changes were observed, and the effect of HSP70 on cell migration ability was detected via the wound healing. The results demonstrated that GGA at 50 and 200 μmol/L could significantly induce HSP70 expression in A549 cells (P < 0.05). Furthermore, HSP70 induced by 200 μmol/L GGA significantly inhibited the changes of E-cadherin, vimentin, and cell morphology induced by TGF-β (P < 0.05), while HSP70 induced by 50 μmol/L GGA did not. The results of the wound healing assay indicated that 200 μmol/L GGA significantly inhibited A549 cell migration induced by TGF-β. Taken together, the results of the present study demonstrated that overexpression of HSP70 inhibited the TGF-β induced EMT process and changed the cell morphology and migratory ability induced by TGF-β in A549 cells.     

Retracted: Upregulation of microRNA-140-3p inhibits epithelial-mesenchymal transition,invasion, and metastasis of hepatocellular carcinoma through inactivation of the MAPK signaling pathway by targeting GRN

Invasion and metastasis in hepatocellular carcinoma (HCC) results in poor prognosis. Human intervention in these pathological processes may benefit the treatment of HCC. The aim of the present study is to elucidate the mechanism of miR-140-3p affecting epithelial-mesenchymal transition (EMT), invasion, and metastasis in HCC. Microarray analysis was performed for differentially expressed genes screening. The target relationship between miR-140-3p and GRN was analyzed. Small interfering RNA (siRNA) against granulin (GRN) was synthesized. EMT markers were detected, and invasion and migration were evaluated in HCC cells introduced with a miR-140-3p inhibitor or mimic, or siRNA against GRN. A mechanistic investigation was conducted for the determination of mitogen-activated protein kinase (MAPK) signaling pathway-related genes and EMT markers (E-cadherin, N-cadherin, and Vimentin). GRN was highlighted as an upregulated gene in HCC. GRN was a target gene of miR-140-3p. Elevation of miR-140-3p or inhibition of GRN restrained the EMT process and suppressed the HCC cell migration and invasion. HCC cells treated with the miR-140-3p mimic or siRNA-GRN exhibited decreased GRN expression and downregulated the expressions of the MAPK signaling pathway-related genes, N-cadherin, and Vimentin but upregulated the expression of E-cadherin. GRN silencing can reverse the activation of the MAPK signaling pathway and induction of EMT mediated by miR-140-3p inhibition. Taken together, the results show that miR-140-3p confers suppression of the MAPK signaling pathway by targeting GRN, thus inhibiting EMT, invasion, and metastasis in HCC.     

MiR-5047在乳腺癌细胞增殖迁移中的作用及表达调控*

探讨miR-5047在乳腺癌细胞中的表达及其在乳腺癌细胞增殖和迁移中的作用,并明确地西他滨在miR-5047表达调控中的作用。通过实时荧光定量PCR(qRT-PCR)检测人乳腺癌细胞系和正常乳腺上皮细胞MCF10A中miR-5047的表达水平;将miR-5047模拟物(mimic),阴性对照(NC)分别转染至MDA-MB-231和MCF7细胞,经平板克隆实验、MTT实验、划痕愈合实验检测乳腺癌细胞的增殖和迁移能力,通过qRT-PCR和Western blot检测相关基因表达及蛋白水平。使用浓度5 μmol/L和10 μmol/L的地西他滨分别处理MDA-MB-231和MCF-7细胞,经qRT-PCR检测不同浓度和处理时间条件下地西他滨对miR-5047表达的影响。同时,通过形态观察和Western blot检测地西他滨对乳腺癌细胞上皮间质转化的影响。与正常乳腺上皮细胞MCF-10A相比,miR-5047在乳腺癌细胞中表达均显著下调。miR-5047过表达可显著抑制乳腺癌细胞的增殖和迁移,促进上皮细胞标志物E-cadherin的表达,抑制间质细胞标志物Vimentin的表达。不同浓度地西他滨处理MDA-MB-231和MCF7细胞后,miR-5047表达均增强,且10 μmol/L作用48 h效果最显著。地西他滨可诱导MDA-MB-231细胞向上皮样转变。miR-5047在乳腺癌细胞系中表达显著下调,过表达miR-5047可抑制乳腺癌细胞的增殖和迁移,地西他滨可促进乳腺癌细胞中miR-5047的表达,并诱导细胞向上皮样转变。