Effect of poly(dimethylsiloxane)-block-poly(oligo (ethylene glycol) methacrylate) amphiphilic block copolymers on dermal fibroblast viability and proliferation

Abstract

In this work, well-defined poly(dimethylsiloxane)-b-poly(oligo (ethylene glycol) methacrylate) (PDMS-b-POEGMA) amphiphilic block copolymers were synthesized and their effect on human dermal fibroblast were investigated. Anionic ring opening polymerization (ROP) and atom transfer radical polymerization (ATRP) were used to synthesis the block copolymers. The molecular weight of synthesized copolymers ranged from 1000 to 2300?Da by changing the number of both PDMS and POEGMA units. It was found that the copolymer having low molecular weight decreased the fibroblast viability and proliferation by inducing apoptosis. It was proved by flow cytometry and TUNEL assay that human dermal fibroblast experienced apoptosis after exposure to synthesized amphiphilic copolymers. The results of this work suggest the use of PDMS-b-POEGMA amphiphilic copolymers with low molecular weight for hypertrophic scars remediation.     

Altered dermal fibroblast behavior in a collagen V haploinsufficient murine model of classic Ehlers–Danlos syndrome

Mutations in collagen V are associated with classic Ehlers–Danlos syndrome (EDS). A significant percentage of these mutations result in haploinsufficiency for collagen V. The purpose of this work was to determine if changes in collagen V expression are associated with altered dermal fibroblast behavior contributing to the poor wound healing response. A haploinsufficient Col5a1^+/? mouse model of EDS was utilized. In vivo wound healing studies demonstrated that mutant mice healed significantly slower than Col5a1^+/+ mice. The basis for this difference was examined in vitro using dermal fibroblast strains isolated from Col5a1^+/? and Col5a1^+/+ mice. Fibroblast proliferation was determined for each strain by counting cells at different time points after seeding as well as using the proliferation marker Ki-67. Fibroblast attachment to collagens I and III and fibronectin also was analyzed. In addition, in vitro scratch wounds were used to analyze fibroblast wound closure. Significantly decreased fibroblast proliferation was observed in Col5a1^+/? compared to Col5a1^+/+ fibroblasts. Our data indicate that the decreased fibroblast number was not due to apoptosis. Wildtype Col5a1^+/+ fibroblasts attached significantly better to components of the wound matrix (collagens I and III and fibronectin) than Col5a1^+/? fibroblasts. A significant difference in in vitro scratch wound closure rates also was observed. Col5a1^+/+ fibroblasts closed wounds in 22 h, while Col5a1^+/? fibroblasts demonstrated ~80% closure. There were significant differences in closure at all time points analyzed. Our data suggest that decreased fibroblast proliferation, extracellular matrix attachment, and migration contribute to the decreased wound healing response in classic EDS.     

增生性瘢痕中时序变化的骨形态发生蛋白及其受体

背景：骨形态发生蛋白与瘢痕的形成和发展相关,而成纤维细胞与瘢痕的增殖、成熟关系密切,但骨形态发生蛋白在不同时期增生性瘢痕成纤维细胞中的表达变化尚少见报道。 目的：检测骨形态发生蛋白2/4、骨形态发生蛋白7、骨形态发生蛋白IA型受体在不同时期增生性瘢痕成纤维细胞中的表达水平。 方法：应用免疫组织化SP法检测来自2008年3月至2010年7月广西壮族自治区人民医院美容整形外科患者的增殖期增生性瘢痕和成熟期增生性瘢痕中成纤维细胞中骨形态发生蛋白2/4、骨形态发生蛋白7、骨形态发生蛋白IA型受体的表达。 结果与结论：增生性瘢痕增殖期成纤维细胞中骨形态发生蛋白2/4的表达水平显著高于成熟期(P < 0.05);但不同时期增生性瘢痕成纤维细胞中骨形态发生蛋白7和骨形态发生蛋白IA型受体的表达差异无显著性意义  (P > 0.05)。表明在增生性瘢痕由增生期至成熟期的发展过程中,其成纤维细胞中的骨形态发生蛋白2/4的表达下调,而骨形态发生蛋白7、骨形态发生蛋白IA型受体的表达无变化。 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程全文链接：     

蛇床子素干预人增生性瘢痕成纤维细胞增殖及转化生长因子β1的表达

背景：蛇床子素对体外培养人增生性瘢痕成纤维细胞增殖和细胞分泌的转化生长因子β1有抑制作用,但其具体作用机制尚待进一步研究。 目的：体外观察蛇床子素对人增生性瘢痕成纤维细胞增殖以及对细胞转化生长因子β1的影响。 方法：体外原代培养人增生性瘢痕成纤维细胞,以不同浓度的蛇床子素作用于成纤维细胞,观察细胞形态的变化,应用MTT法和生长曲线法检测蛇床子素对细胞增殖活性的影响。免疫组织化学检测细胞转化生长因子β1的表达。 结果与结论：蛇床子素能明显抑制人增生性瘢痕成纤维细胞的生长。MTT法检测的IC50为(15.2±2.0) μmol/L,可以明显下调细胞转化生长因子β1的表达(P < 0.05)。说明蛇床子素对人增生性瘢痕成纤维细胞有很强的生长抑制作用并可以下调转化生长因子β1的表达。     

Red blood cells inhibit proliferation and stimulate apoptosis in human lung fibroblasts in vitro

Cell proliferation and apoptosis are both important mechanisms for the regulation of tissue homeostasis. For instance, proliferation is crucial in wound repair, whereas apoptosis is important for removal of damaged cells and resolution of inflammation. Imbalance between cell proliferation and apoptosis can therefore lead to pathological conditions and disease. In inflammatory and fibrotic lung disorders, red blood cells (RBCs) can interact with fibroblasts and connective tissue. In the present study, we therefore hypothesized that the presence of RBCs can affect fibroblast proliferation and apoptosis. Human foetal lung fibroblasts (HFL-1) were cultured in the presence or absence of purified whole RBCs and RBC-conditioned media. RBC significantly decreased fibroblast proliferation as determined both by DNA content analysis (Hoechst 33258 staining, P < 0.01; WST-1, P < 0.001) and BrdU incorporation. After treatment with staurosporine (STS) for 48 h, apoptosis was determined by TUNEL and propidium iodide staining followed by flow cytometry analysis. RBCs augmented STS-induced apoptosis (median: 46.4%; range 12.0-90.4) compared to control cells (median 26.2%; range 7.1-45.5). Thus, our data indicate that the presence of RBCs affects both fibroblast proliferation and susceptibility to undergo apoptosis. Our findings therefore suggest a role for RBCs in regulating fibroblast homeostasis after tissue injury.     

Dextran derivatives modulate collagen matrix organization in dermal equivalent

Dextran derivatives can protect heparin binding growth factor implied in wound healing, such as transforming growth factor-β1 (TGF-β1) and fibroblast growth factor-2 (FGF-2). The first aim of this study was to investigate the effect of these compounds on human dermal fibroblasts in culture with or without TGF-β1. Several dextran derivatives obtained by substitution of methylcarboxylate (MC), benzylamide (B) and sulphate (Su) groups were used to determine the effects of each compound on fibroblast growth in vitro. The data indicate that sulphate groups are essential to act on the fibroblast proliferation. The dextran derivative LS21 DMCBSu has been chosen to investigate its effect on dermal wound healing process. Fibroblasts cultured in collagenous matrices named dermal equivalent were treated with the bioactive polymer alone or associated to TGF-β1 or FGF-2. Cross-sections of dermal equivalent observed by histology or immunohistochemistry, demonstrated that the bioactive polymer accelerates the collagen matrices organization and stimulates the human type-III collagen expression. This bioactive polymer induces apoptosis of myofibroblast, property which may be beneficial in treatment of hypertrophic scar. Culture media analyzed by zymography and Western blot showed that this polymer significantly increases the secretion of zymogen and active form of matrix metalloproteinase-2 (MMP-2), involved in granulation tissue formation. These data suggest that this bioactive polymer has properties which may be beneficial in the treatment of wound healing.     

骨髓间充质干细胞条件培养液对瘢痕成纤维细胞生物活性的影响

背景：间充质干细胞移植可通过多种调节机制促进皮肤创伤修复,抑制瘢痕形成,目前多数学者认为间充质干细胞分泌的生物活性因子起主要作用。 目的：观察骨髓间充质干细胞条件培养液对增生性瘢痕成纤维细胞增殖能力及胶原合成的影响。 方法：分离培养人骨髓间充质干细胞和增生性瘢痕成纤维细胞,制备骨髓间充质干细胞条件培养液。将12,24,48 h收集的条件培养液孵育体外培养的增生性瘢痕成纤维细胞24 h,并与空白对照组比较,采用CCK-8实验检测增生性瘢痕成纤维细胞的增殖活性,Real-time PCR检测增生性瘢痕成纤维细胞中Ⅰ型和Ⅲ型胶原的表达。 结果与结论：与空白对照组相比较,在24 h和48 h收集的骨髓间充质干细胞条件培养液对增生性瘢痕成纤维细胞的增殖具有明显的抑制作用(P < 0.01),对其胶原的合成也具有显著的抑制作用(P < 0. 01)。结果表明骨髓间充质干细胞条件培养液可通过分泌抗纤维化的生物活性因子抑制增生性瘢痕成纤维细胞的增殖和胶原产生,为细胞疗法策略减轻瘢痕提供了新的理论支持。 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程全文链接：     

The CC chemokine eotaxin/CCL11 has a selective profibrogenic effect on human lung fibroblasts

BACKGROUND: Eotaxin/CCL11 plays an important role in asthma. It acts through the chemokine receptor CCR3 expressed on hematopoietic and nonhematopoietic cells in the lung. OBJECTIVE: To determine whether eotaxin/CCL11 modulates lung and bronchial fibroblast properties and thereby might contribute to airway remodeling. METHODS: CCR3 expression was characterized on a lung fibroblast line (MRC-5; flow cytometry, fluorescent microscopy, RT-PCR, and Northern blotting), on primary bronchial fibroblasts (flow cytometry), and on fibroblasts in human lung tissue (confocal laser microscopy). The effects of eotaxin/CCL11 on lung fibroblast migration (Boyden chamber), proliferation (tritiated thymidine incorporation), alpha-smooth muscle actin expression (ELISA), 3-dimensional collagen gel contraction (floating gel), pro-alpha1(I) collagen mRNA (Northern blotting), total collagen synthesis (tritiated proline incorporation), matrix metalloproteinase activity (gelatin zymography), and TGF-beta(1) release (ELISA) were evaluated. The contribution of eotaxin/CCL11/CCR3 binding on lung fibroblasts was also investigated by neutralizing experiments. RESULTS: CCR3 is constitutively expressed in cultured lung and primary bronchial fibroblasts and colocalizes with specific surface markers for human fibroblasts in lung tissue. Eotaxin/CCL11 selectively modulates fibroblast activities by increasing their proliferation, matrix metalloproteinase 2 activity, and collagen synthesis but not their differentiation into myofibroblasts, contractility in collagen gel, or TGF-beta(1) release. Eotaxin/CCL11 enhances migration of lung fibroblasts in response to nonspecific chemoattractants, and this effect is completely inhibited by anti-CCR3-neutralizing antibodies. CONCLUSION: These data demonstrate that eotaxin/CCL11 has a direct and selective profibrogenic effect on lung and bronchial fibroblasts, providing a novel mechanism whereby eotaxin/CCL11 can participate in airway remodeling in asthma.     

Enhanced expression of caspase-3 in hypertrophic scars and keloid: induction of caspase-3 and apoptosis in keloid fibroblasts in vitro

Recent studies have suggested that the regulation of apoptosis during wound healing is important in scar establishment and development of pathological scarring. To examine the phenomenon of apoptosis and its involvement in the process of pathological scarring, we immunohistochemically quantified differential levels of expression of caspase-3 and -2, which are activated during apoptosis in vitro, in surgical resected scar tissues. We divided 33 cases of normally healed flat scars and 18 cases of pathological scars (15 cases of hypertrophic scars and 3 cases of keloid) into three groups (S1 = <10 months' duration; S2 = 10 to 40 months' duration; and S3 = >40 months' duration) according to the duration of scar. In all three groups examined, the semiquantitative scores for caspase-3 staining were significantly higher for the combination of hypertrophic scars and keloid as a group compared with normally healed flat scars, suggesting reduced cell survival and increased apoptotic cell death in hypertrophic scars and keloid. Apoptosis and caspase proteolytic activities were examined in vitro using two flat scar-derived fibroblast lines (FSFB-1 and -2) and two keloid-derived fibroblast lines (KFB-1 and -2). After 24 hours of serum deprivation, apoptotic cells were significantly increased in both KFB lines, whereas serum deprivation of FSFB-1 cells did not result in a significant increase in apoptotic cell number. After serum deprivation, significant increases in caspase-3 proteolytic activities were detected in both KFB lines compared with both FSFB lines. In contrast, no significant differences with caspase-8 activity were observed between similarly treated KFB and FSFB lines. Furthermore, serum deprivation-induced apoptosis of KFB-2 cells was significantly inhibited by the caspase-3 inhibitor Ac-Asp-Glu-Val-Asp-fluoromethyl ketone (DEVD-FMK), indicating that caspase-3 is important for serum deprivation-induced apoptosis in KFB-2 cells. Considering the role of caspase-3 as a key effector molecule in the execution of apoptotic stimuli, our results suggested that enhanced expression of caspase-3 in hypertrophic scars and keloid induces apoptosis of fibroblasts, which may play a role in the process of pathological scarring.     

增生性瘢痕成纤维细胞中CyclinD1，CDK2，CDK4作用及与 细胞周期的相关性

背景：增生性瘢痕的发生和发展可能与细胞周期调节相关基因的异常表达有关。 目的：检测增生性瘢痕不同阶段细胞CyclinD1、CDK2和CDK4 mRNA及蛋白的表达,以及同一标本增生性瘢痕成纤维细胞的细胞周期运行中3者之间的一致性。 方法：采集32份增生性瘢痕标本,以增生性瘢痕形成时段分为3个月组、6个月组、1年组、2年组,各8份,并以8份正常真皮标本作为对照。利用实时荧光定量PCR法、Western blotting法检测标本中CyclinD1、CDK2、CDK4 mRNA及蛋白表达,流式细胞术检测成纤维细胞的细胞周期。 结果与结论：增生性瘢痕标本发展过程中CyclinD1、CDK2、CDK4 mRNA及蛋白表达水平由强至弱变化。3,6个月增生性瘢痕中成纤维细胞主要分布在S、G2 /M期;1,2年增生性瘢痕与对照组成纤维细胞多处于G0/G1期。表明增生性瘢痕不同时期CyclinD1、CDK2、CDK4各自的 mRNA和蛋白表达趋势基本一致,同时增生性瘢痕中成纤维细胞的细胞周期分布与这3个蛋白所执行的功能情况相对应。     

磷脂多不饱和脂肪酸抑制兔耳增生性瘢痕

背景：多不饱和脂肪酸有抑制细胞炎症反应及免疫功能的作用,增生性瘢痕的形成与炎症、细胞免疫、细胞因子有着密切关系,但目前尚无应用多不饱和脂肪酸防治增生性瘢痕的实验研究。 目的：探讨磷脂多不饱和脂肪酸对兔耳增生性瘢痕的抑制作用。 方法：在9只新西兰大白兔兔耳腹侧做直径1 cm的圆形全层皮肤缺损创面,每侧6个,共108个,其中形成增生性瘢痕92个,瘢痕形成率为85%。实验分3组：每只兔耳靠前3个创面涂磷脂多不饱和脂肪酸霜,右耳靠后3个创面涂多磺酸黏多糖乳膏,创面上皮化后立即涂药,每日1次,左耳靠后3个创面自然愈合。分别在术后28,42,63,90 d,观察创面的愈合情况;显微镜下观察瘢痕组织的厚度、胶原纤维和成纤维细胞密度;免疫组织化学染色检测胶原纤维的表达。 结果与结论：涂抹磷脂多不饱和脂肪酸霜和多磺酸黏多糖乳膏可使增生性瘢痕体积缩小、厚度变薄、成纤维细胞密度减小、胶原纤维表达减少。尤以磷脂多不饱和脂肪酸霜的效果最为明显。说明磷脂多不饱和脂肪酸可抑制兔耳增生性瘢痕的形成,减轻瘢痕的增生程度。     

饥饿诱导增生性瘢痕成纤维细胞自噬发生

 ［摘要］目的:探讨饥饿诱导的增生性瘢痕成纤维细胞自噬的发生。方法:原代培养增生性瘢痕成纤维细胞至对数生长期,用EBSS（Earle’s balanced salt solution）代替培养液,分别饥饿0 h、1 h、2 h、3 h,应用Western blotting和定量PCR(qRT-PCR)方法分别检测成纤维细胞微管相关蛋白1轻链3（microtubule-associated protein 1 light chain 3, LC-3）和自噬相关蛋白Beclin-1的蛋白和mRNA表达,单丹磺酰戊二胺(monodansylcadaverine,MDC)染色和透射电镜观察自噬体。结果:Western blotting和qRT-PCR 检测LC3和Beclin-1 蛋白和mRNA表达在饥饿1 h开始增加,2 h达到高峰,3 h逐渐下降,均明显大于对照组（P<0.05）。荧光显微镜和电镜下观察到饥饿2 h诱导的成纤维细胞出现自噬囊泡。结论: 饥饿可诱导增生性瘢痕成纤维细胞发生自噬,可能与增生性瘢痕的形成有关。     

rhIL-17A对人皮肤角质形成细胞和成纤维细胞活性和凋亡的影响

目的: 研究重组人白细胞介素17A (rhIL-17A)对人角质形成细胞和成纤维细胞活性、凋亡及成纤维细胞促纤维化因子分泌的影响,探讨IL-17在系统性硬皮病发病中的作用。方法: 用不同浓度的rhIL-17A作用于角质形成细胞和成纤维细胞,CCK-8测定其对角质形成细胞和成纤维细胞活性的影响;Western blotting 检测核转录因子NF-κB/p65及其抑制性蛋白IκBα的表达量;流式细胞术观察细胞凋亡变化;ELISA 检测rhIL-17A 处理组和对照组成纤维细胞培养液上清中IL-6和TGF-β₁的含量。结果: rhIL-17A处理组和对照组角质形成细胞活性没有明显差别;rhIL-17A 处理组成纤维细胞活性较对照组明显升高(P<0.05),rhIL-17A 处理组成纤维细胞NF-κB/p65表达量增加,而IκBα表达量降低;rhIL-17A 对2种细胞凋亡率无明显影响;rhIL-17A 处理组成纤维细胞IL-6 和TGF-β₁分泌量增加。结论: rhIL-17A对体外培养人角质形成细胞的活性没有明显影响,却可以显著增强体外成纤维细胞的活性,IL-17A对成纤维细胞活性增强的这种促进作用,可能是通过激活NF-κB实现。rhIL-17A可能通过刺激成纤维细胞分泌IL-6 和TGF-β₁,从而进一步引起成纤维细胞增殖和胶原合成。IL-17可能参与了系统性硬皮病的发病过程。     

黄芩苷抑制CA46细胞增殖和诱导凋亡的作用机制探讨

目的： 研究中药黄芩苷（baicalin）对人Burkitt淋巴瘤细胞株CA46细胞增殖、凋亡的影响并探讨其可能作用机制。 方法：应用MTT法绘制细胞生长曲线观察黄芩苷对CA46细胞增殖的影响;Annexin V-FITC/PI双染流式细胞术、细胞DNA片段化、TdT酶介导的原位缺口末端标记(TUNEL)法检测黄芩苷诱导CA46细胞凋亡的能力;RT-PCR法检测黄芩苷作用前后c-myc、bcl-2 mRNA表达水平的变化,Western blotting法检测c-Myc、Bcl-2、procaspase-3(caspase-3前体)、PARP(多聚ADP核糖聚合酶)蛋白水平的变化。结果：细胞生长曲线结果显示黄芩苷能明显抑制CA46细胞增殖,半数抑制浓度(IC50)约为10 μmol/L;Annexin V-FITC/PI双染流式细胞术早期凋亡的检出、TUNEL晚期凋亡细胞的检出和细胞DNA片段化凋亡梯带的检出,均证实黄芩苷能有效诱导CA46细胞凋亡,细胞凋亡率呈现浓度依赖性递增。黄芩苷作用后CA46细胞c-myc、bcl-2 mRNA和c-Myc、Bcl-2、procaspase-3、PARP(116 kD)蛋白的表达呈现时间依赖性递减,而PARP(85 kD)表达呈现时间依赖性递增。结论：黄芩苷能有效抑制CA46细胞增殖,诱导其凋亡;c-Myc、Bcl-2表达水平下调和caspase-3激活可能参与了这一作用过程。     

Gene expression profiles from hypertrophic scar fibroblasts before and after IL-6 stimulation

The structural rearrangement of collagen fibres in hypertrophic scar causes abnormal contracture, low tensile strength, and raised scars, which cause functional impairment and disfigurement. It is hypothesized that changes in the genes of cytokines, extracellular matrix proteins, and proteins regulating programmed cell death are related to hypertrophic scar formation. To test this hypothesis, fibroblasts were cultured from hypertrophic scars and their response to interleukin-6 (IL-6) stimulation was studied by defining their gene expression profiles. Affymetrix gene chip analysis was used to identify up- or down-regulation in the 12 625 genes present in the affymetrix array. RT-PCR and ELISA assays were used to validate microarray expression profiles further. Comparison of gene profiles showed an increase of 12 genes in hypertrophic scar fibroblasts compared with normal skin fibroblasts, while the expression of 14 genes decreased. Thirty-three genes were affected by IL-6 treatment in the hypertrophic scar fibroblasts, while 57 genes were affected in normal skin fibroblasts. Messenger RNA to beta-actin ratios for matrix metalloproteinase-1 (MMP-1) and MMP-3 were increased with IL-6 in normal skin fibroblasts from 2.43 +/- 0.06 to 5.50 +/- 0.45 and from 0.75 +/- 0.09 to 1.98 +/- 0.01, respectively. No change in these matrix metalloproteinases could be shown with IL-6 stimulation in hypertrophic scar fibroblasts. Secreted protein levels of pro-MMP-1 and MMP-3 were elevated in the supernatants from normal skin fibroblasts from 2.00 +/- 0.09 and 1.72 +/- 0.10 ng/ml to 4.60 +/- 0.12 and 3.41 +/- 0.20 ng/ml, respectively, after treatment with IL-6 (p < 0.05). No changes were observed in hypertrophic scar fibroblasts treated with IL-6. Values are means +/- SEM. The absence of any up-regulation of MMP-1 and MMP-3 in hypertrophic scar fibroblasts, in response to IL-6, suggests that suppression of matrix metalloproteinases may play a role in the excessive accumulation of collagen formed in hypertrophic scars. While the pathogenesis of abnormal hypertrophic scars remains poorly understood, the use of gene expression arrays may prove helpful in identifying the mechanisms responsible for this type of abnormal scar formation and in formulating an effective therapeutic protocol.     

小干扰RNA沉默hPTTG1基因对卵巢癌细胞增殖和凋亡的影响

目的: 利用RNA干扰技术沉默人垂体瘤转化基因1( hPTTG1 )表达,观察卵巢癌细胞增殖能力和细胞凋亡的改变并探讨分子机制。方法: 化学合成靶定 hPTTG1 的小干扰RNA(siRNA)转染体外培养的A2780细胞,RT-PCR和Western blotting检测 hPTTG1 及c-myc表达水平;MTT法和 -TdR掺入实验检测 hPTTG1 对细胞增殖的影响;annexin V/PI染色流式细胞术和TUNEL法测定各组细胞凋亡情况。结果: hPTTG1 siRNA转染组 hPTTG1 mRNA和蛋白表达下降,抑制率分别为70.5%±3.9%和63.8%±4.5%;转染 hPTTG1 siRNA 24 h细胞吸光度值开始下降,48 h抑制效果最明显,抑制率为42.9%±5.2%;转染 hPTTG1 siRNA后细胞 -TdR放射性计数明显低于空白组和阴性对照组; hPTTG1 siRNA干扰组细胞存活率下降,细胞凋亡率和坏死率均增加;TUNEL染色分析结果可见 hPTTG1 siRNA干扰组凋亡指数明显高于空白组和阴性对照组; hPTTG1 干扰后c-myc mRNA和蛋白表达均下调。结论: hPTTG1 siRNA通过下调c-myc表达抑制A2780细胞增殖,诱导细胞凋亡, hPTTG1 可作为卵巢癌基因治疗的候选靶点。     

miR-425 regulates cell proliferation,migration and apoptosis by targeting AMPH-1 in non-small-cell lung cancer

BackgroundMicroRNAs (miRNAs) have been proved to act as vital roles on non-small-cell lung cancer (NSCLC), and miR-425 has been proven to serve an important function in several tumors. However, the functional role of miR-425 on NSCLC is still unclear.MethodsThe mRNA and protein expression of miR-425 and AMPH-1 were determined by qRT-PCR and western blot analysis, respectively. NSCLC cells (SK-MES-1 and A549) proliferation and migration were measured by CCK-8 and transwell assay, respectively. Cell apoptosis was assessed by flow cytometry and western blotting, In addition, luciferase reporter assay was carried out to confirm the direct targeting of AMPH-1 by miR-425. Xenograft experiments were performed to observe the tumorigenesis of miR-425 in vivo.ResultsThe results showed that miR-425 was overexpressed and AMPH-1 expression was downregulated in SK-MES-1 and A549 cells. Silencing miR-425 inhibited proliferation, migration and promoted apoptosis of NSCLC cells. Moreover, we proved that miR-425 could target AMPH-1. The expression of AMPH-1was upregulated in A549 with miR-425 inhibitor. Moreover, miR-425 knockdown were less tumorigenic than the control in vivo.ConclusionsTaken together, miR-425 could promote the proliferation, invasion and suppress apoptosis by targeting AMPH-1 in NSCLC cells. miR-425/AMPH-1 axis may represent a potential therapeutic strategy or novel prognostic biomarkers to NSCLC.