miR-502-3p 通过靶向结合 CBL 抑制卵巢癌增殖并诱导凋亡

目的 探究微小 RNA 502-3p (micro RNA 502-3p, miR-502-3p) 通过靶向结合 Casitas B 细胞淋巴 瘤 (Casitas B-cell lymphoma, CBL) 参与卵巢癌增殖和凋亡的机制。 方法 下载 GSE66957、 GSE119056、 TCGA_ OV 卵巢癌相关数据矩阵, 分析 miR-502-3p、 CBL 与卵巢癌的关系; 构建过表达 miR-502-3p、 CBL 的 SKOV3 和 HO8910 细胞系, 分别采用细胞计数试剂盒 ( cell counting kit 8, CCK-8)、 克隆形成实验、 流 式细胞术检测细胞增殖和凋亡情况; 通过荷瘤裸鼠实验, 观察过表达 CBL 对肿瘤生长的影响; 验证 miR502-3p 与 CBL 的靶向关系。 结果 生物信息学分析显示, 卵巢癌组织中 CBL 水平高于癌旁组织, miR-502- 3p 水平低于癌旁组织, CBL 水平与患者预后、 细胞增殖基因表达有关 (P< 0. 05)。 miR-502-3p 与 CBL 存在 靶向关系, 与 Vector 组比较, CBL 组肿瘤的体积及重量增加 (P< 0. 05); 与 miR-NC 组比较, miR-502-3p 组 SKOV3、 HO8910 细胞中 CBL 蛋白表达、 细胞活力、 克隆数降低, 细胞凋亡率升高 (P< 0. 05), 但 CBL 可逆转上述细胞变化。 结论 miR-502-3p 可通过靶向下调 CBL 抑制卵巢癌细胞的增殖, 并诱导其凋亡。     

Mechanism underlying the regulation of lncRNA ACTA2-AS1 on CXCL2 by absorbing miRNA-532-5p as ceRNA in the development of ovarian cancer

Objective: To explore the mechanism underlying the regulation of long non-coding RNA (LncRNA) ACTA2-AS1 on CXCL2 as a ceRNA of miR-532-5p in the progression of ovarian cancer (OC). Methods: A qRT-PCR assay was carried out for analyzing the expression changes of ACTA2-AS1, miR-532-5p, as well as CXCL2 in OC tissues and corresponding healthy paracancerous tissues HOSEpiC (human ovarian epithelial cells), and OC cells. OC cells were grouped and transfected, and the fluorescent in situ hybridization was adopted for evaluating ACTA2-AS1 in the cells. Additionally, a dual luciferase reporter (DLR) assay was carried out for verifying the correlation of ACTA2-AS1 with miR-532-5p and of miR-532-5p with CXCL2. Cells were transfected with si-ACTA2-AS1, miR-532-5p, or CXCL2 overexpression plasmids, and then the cell proliferation, invasion, and apoptosis were determined using MTT, Transwell, and flow cytometry assays, respectively. Results: Compared with paracancerous tissues and HOSEpiC cells, OC tissues and cells showed increased ACTA2-AS1 and CXCL2 expression and decreased miR-532-5p expression (all P<0.05). ACTA2-AS1 acted as ceRNA in OC by negatively regulating miR-532-5p. Additionally, upregulating ACTA2-AS1 intensified the proliferation and invasion of cancer cells and suppressed their apoptosis (all P<0.05), and inhibition of it resulted in opposite results. In contrast, overexpressing miR-532-5p suppressed the proliferation, invasion, and clone formation of the cells and promoted their apoptosis (all P<0.05). The effect of ACTA2-AS1 on OC cells can be partially reversed by overexpressing miR-532-5p. Moreover, CXCL2, positively correlated with ACTA2-AS1 in expression (P<0.0001, r=0.7385), was the target of miR-532-5p, and its overexpression could partially offset the influence of miR-532-5p on OC cells. Conclusion: LncRNA ACTA2-AS1 can act as a tumor promoter in OC by absorbing miR-532-5p as ceRNA and regulating CXCL2, and ACTA2-AS1 inhibitor is expected to play a role in targeted therapy of OC.     

长链非编码RNA UCA1 靶向miR-582-5p 对膀胱癌UM-UC-3细胞生存和运动能力的调节作用及机制研究

目的：探究长链非编码RNA-UCA1通过靶向miR-582-5p对膀胱癌细胞生存和运动能力的作用及作用机制。方法：用UCA1-shRNA(sh-UCA1)和(或)miR-582-5p inhibitor转染细胞,荧光定量检测转染效率及miR-582-5p的表达水平;荧光素酶报告实验确定UCA1和miR-582-5p的靶向关系;CCK8检测细胞活性,流式检测细胞凋亡情况,侵袭及划痕实验检测细胞侵袭迁移能力,免疫印迹检测细胞增殖、凋亡及迁移相关蛋白的表达。结果：sh-UCA1能显著降低膀胱癌细胞UCA1表达水平(P<0.05),促进miR-582-5p表达(P<0.05);miR-582-5p-inhibitor能明显减弱sh-UCA1对miR-582-5p表达的促进作用(P<0.05);荧光素酶报告实验表明UCA1上有miR-582-5p的结合位点;沉默UCA1可显著抑制膀胱癌细胞增殖及Ki67的表达,促进细胞凋亡及cleaved caspase-3的表达(P<0.05);同时,sh-UCA1还能显著抑制膀胱癌细胞侵袭、迁移及VEGF的表达(P<0.05);此外,miR-582-5p inhibitor可显著减弱sh-UCA1对细胞增殖、凋亡及侵袭迁移能力的作用(P<0.05)。结论：UCA1可通过靶向miR-582-5p增强膀胱癌UM-UC-3细胞的生存及运动能力。     

circ-SFMBT2通过靶向miR-7-5p/ADAM10轴对非小细胞肺癌细胞生物学行为的影响CSCD

目的探讨circ-SFMBT2对非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞生物学行为的影响以及对miR-7-5p/ADAM10分子轴的调控作用。方法﹑采用qRT-PCR法与Western印迹法分别检测NSCLC与癌旁组织中circ-SFMBT2、miR-7-5p、ADAM10的表达量;用Pearson法分析circ-SFMBT2与miR-7-5p,以及miR-7-5p与ADAM10的相关性;体外培养人支气管上皮样细胞(human bronchial epithelial-like cells,HBE)与肺癌细胞系H1650、H460、A549、H1299。用CCK-8与EdU实验检测细胞的增殖能力。用平板克隆形成实验检测细胞的克隆形成能力。用流式细胞术检测细胞凋亡率。用Transwell小室实验检测细胞侵袭。用双荧光素酶报告实验检测circ-SFMBT2与miR-7-5p、以及miR-7-5p与ADAM10的靶向关系。用裸鼠移植瘤实验检测敲低circ-SFMBT2对移植瘤生长的影响。用免疫组化实验检测移植瘤组织中ADAM10与Ki67蛋白阳性率。结果circ-SFMBT2与ADAM10在NSCLC组织及细胞系中表达升高,而miR-7-5p的表达降低,circ-SFMBT2与miR-7-5p的表达呈负相关,而miR-7-5p与ADAM10的表达呈负相关。沉默circ-SFMBT2及miR-7-5p过表达可抑制细胞增殖、克隆形成及侵袭,还可促进其凋亡。circ-SFMBT2可靶向调控miR-7-5p,而ADAM10是miR-7-5p的靶基因。沉默circ-SFMBT2与抑制miR-7-5p联合作用,以及miR-7-5p过表达与ADAM10过表达联合作用均可促进细胞增殖、克隆形成及侵袭,并抑制其凋亡。沉默circ-SFMBT2可抑制移植瘤的生长。结论︰沉默circ-SFMBT2可通过调控miR-7-5p/ADAM10分子轴而减弱NSCLC细胞增殖、克隆形成,侵袭能力并诱导其凋亡。     

MicroRNA-490-5p is a novel tumor suppressor targeting c-FOS in human bladder cancer

Introduction

Recent studies have demonstrated the critical roles of micro-RNAs in tumorigenesis and tumor progression. Here, we describe the regulation and function of miR-490-5p in bladder cancer.

Material and methods

Paired tissue samples were collected from bladder cancer patients (n = 20). Real-time PCR revealed that miR-490-5p expression was significantly down-regulated in human bladder cancer tissues and cells. Also there was an inverse relationship between the expression level of miR-490-5p and the pathological grade of bladder cancer. Western blotting was performed to detect the expression levels of c-FOS and TET1 in 6 matched tumor tissue samples and 4 bladder cell lines. Furthermore, to better understand the underlying mechanisms of miR-490-5p, we conducted gain and loss of function analysis by transfecting bladder cancer T24 cells with chemically synthesized miR-490-5p mimics and inhibitor, respectively.

Results

We found that overexpression of miR-490-5p in T24 cells could inhibit cell proliferation and invasion and induce cell apoptosis. Conversely, suppression of miR-490-5p expression induced cell proliferation and invasion, while it inhibited cell apoptosis. In addition, our bioinformatics prediction and experimental data showed that c-FOS was a potential target of miR-490-5p. The expression level of c-FOS was significantly decreased after miR-490-5p overexpression and significantly increased after miR-490-5p suppression, indicating that c-FOS was a target of miR-490-5p.

Conclusions

These findings suggest that miR-490-5p is a novel tumor suppressor, contributing to the carcinogenesis of bladder cancer by targeting c-FOS.     

miR-495-3p / WIF1 / Wnt 信号通路轴调节视网膜母细胞瘤细胞 增殖、 迁移和侵袭

目的 分析 miR-495-3p / Wnt 抑制因子 (Wnt inhibitor factor 1, WIF1) / Wnt 信号通路轴调节视网 膜母细胞瘤 (retinoblastoma, RB) 细胞增殖、 迁移和侵袭。 方法 生物信息学分析 WIF1 的差异表达及与 临床不良表型之间的关系, 并预测 miRNA 靶标; 双荧光酶素报告实验验证。 构建稳定过表达 WIF1 的 RB 细胞 (Vector 组、 WIF1 组)。 构建稳定过表达 miR-495-3p 的 RB 细胞 (miR-NC 组、 miR-495-3p 组和 miR495-3p + WIF1 组)。 Western 印迹检测 WIF1 蛋白及 Wnt 通路相关蛋白表达, 并进行体外功能试验。 裸鼠移 植瘤实验分析移植瘤体积重量。 免疫组化分析 WIF1 在 RB 组织中水平。 结果 WIF1 低表达, 与迁移、 侵袭 有关, 为 miR-495-3p 靶标, 双荧光酶素报告实验证实 WIF1 是 miR-495-3p 作用靶点。 WIF1 过表达抑制细胞增 殖、 迁移和侵袭, 影响细胞周期, 诱导凋亡。 体内实验显示 WIF1 在 RB 组织中过表达, 其过表达抑制瘤体生 长。 过表达 WIF1 下调 β-catenin、 c-Myc 蛋白水平 (P< 0. 05)。 过表达 miR-495-3p 下调 WIF1 蛋白水平, 上调 β-catenin 和 c-Myc 蛋白水平, 促进细胞增殖迁移、 迁移和侵袭, 抑制细胞凋亡 (P< 0. 05), 增加 WIF1 表达 可逆转 miR-495-3p 的作用 (P< 0. 05)。 结论 miR-495-3p 靶向 WIF1 通过 Wnt 信号通路抑制 RB 细胞增殖、 迁移和侵袭, 并诱导凋亡。     

MiR-124-3p inhibits the migration and invasion of Gastric cancer by targeting ITGB3

BACKGROUNDGastric cancer is one of the major malignant tumors in the world. Integrins expressed in cancer cells can promote tumor progression and migration. MiRNAs can inhibit the expression of target genes by directly binding to their mRNAs and can affect various important biological processes. The aim of this study was to investigate the role of miR-124- 3p and ITGB3 in gastric cancer.METHODSRT-PCR and western blot are used to detect the expression of miR-124-3p, ITGB3 and integrin β3 in gastric cancer tissues and cells. The wound healing, CCK-8 assay, transwell migration and invasion assay were performed to determine the cell proliferation, migration and invasion. What’s more, bioinformatics prediction and luciferase assay was conducted to demonstrated the binding efficiency between miR-124-3p and ITGB3.RESULTSWe verified that ITGB3 and miR-124-3p changes the migration and invasion of gastric cancer cells in vitro. The overexpression or silencing of miR-124-3p inhibited or promoted the proliferation, migration and invasion of both selected gastric cancer cells, and ITGB3 is just the reverse. Meanwhile, we validated that ITGB3 is the target of miR-124-3p by bioinformatics prediction and luciferase assay. Lastly, the expression of ITGB3 in 40 pairs of gastric cancer tissues were significantly higher than that in the adjacent normal tissues, while the expression level of miR-124-3p was significantly decreased in cancer tissues.CONCLUSIONSmiR-124-3p inhibits the migration and invasion of Gastric cancer by targeting ITGB3 in gastric cancer cells. Our results suggested that miR-124-3p and ITGB3 may reasonably serve as a promising therapeutic target.     

miR-130b-3p有望作为人膀胱癌的标志物

目的进一步了解miRNA在膀胱癌中的潜在机制。方法芯片分析4对人膀胱癌组织和相邻正常组织中的miRNA的表达。并用RT-q PCR来验证两个最上调的miRNA及其靶基因的表达是否符合miRNA/mRNA芯片结果。通过相关性分析和双荧光素酶报告实验推断并验证miR-130b-3p可以靶向PTEN。应用CCK8、EDU、流式细胞术、划痕、Transwell和细胞骨架等实验证明miR-130b可以影响膀胱癌细胞的增殖、凋亡、迁移和侵袭。用Western blot检测PI3K/AKT和整合素β1/FAK信号通路的关键靶蛋白。结果人膀胱癌中miR-130b-3p表达高于癌旁且与PTEN表达呈负相关。miR-130b-3p可下调PTEN表达,导致PI3K/AKT和整合素β1/FAK信号通路的激活,且与膀胱癌EJ细胞的增殖、迁移和侵袭相关。细胞转染miR-130b-3p抑制剂时,可以重排细胞骨架。结论本结果揭示miR-130b/PTEN有望用于人膀胱癌诊断和治疗的标志物。     

p53/miR-502-5p/TRAF2通路对骨关节炎软骨细胞损伤的影响

目的:探究微小RNA-502-5p(miR-502-5p)对白细胞介素-1β(IL-1β)诱导的骨关节炎(OA)软骨细胞损伤的影响。方法:Western blot和RT-q PCR检测骨关节炎患者软骨组织和IL-1β诱导的软骨细胞中p53和miR-502-5p的表达;分离培养软骨,并分组处理细胞,分别用MTT法检测细胞活力,流式细胞术检测细胞凋亡,Western blot检测炎性因子和细胞外基质(ECM)相关蛋白的表达,另外,萤光素酶报告实验检测miR-502-5p对p53及肿瘤坏死因子受体相关因子2(TRAF2)的调控作用。结果:与正常软骨组织相比,OA患者软骨组织中miR-502-5p的水平显著降低,p53和TRAF2水平则明显升高(P0.05)。IL-1β处理软骨细胞后,细胞活力下降、细胞凋亡率增加、IL-6、IL-8和TNF-α的蛋白表达水平显著上升;Ⅱ型胶原和蛋白聚糖的蛋白表达显著下调,基质金属蛋白酶(MMP)-3、MMP-9和MMP-13等的蛋白表达显著上调,miR-502-5p mimic转染反转了IL-1β对软骨细胞的这些作用。此外,p53与miR-502-5p之间存在负反馈调节,同时miR-502-5p能够靶向抑制TRAF2的水平。TRAF2沉默同样反转了IL-1β对软骨细胞增殖、凋亡、炎症反应以及ECM相关蛋白表达的影响。结论:调节p53/miR-502-5p/TRAF2通路能够减轻IL-1β诱导的骨关节软骨细胞损伤。     

miR-451 inhibits invasion and proliferation of bladder cancer by regulating EMT

MicroRNAs (miRNAs) have recently been reported play a crucial role in some tumors. In order to investigate the association of miR-451 with bladder cancer, we investigate the expression of miR-451 in bladder cancer tissues and its role in biological behavior of T24, 5637 and J28 bladder cancer cell lines. Quantitative RT-PCR results showed miR-451 was significantly down-regulated in bladder cancer tissues and paracancerous tissues compared with normal bladder tissues. miR-451 expression was significantly associated with histological differentiation degree and TNM stage. Over-expression of miR-451 was established by transfecting miR-451 mimics into T24, 5637 and J28 cells, and its effects on the biological behavior of bladder cancer were studied using transwell assay, migration assay, adhesion assay, MTT and flow cytometry. Results indicated over-expression of miR-451 significantly inhibited cell proliferation, migration, invasion and induced apoptosis of the bladder cancer cells. Furthermore, we investigated the expression level of EMT related proteins in transfected 5637 cells by western blot. Results shown E-cadherin was up-regulated more significantly than N-cadherin, vimentin and Snail. N-cadherin and vimentin were up-regulated significantly when miR-451 was inhibited in miR-451 inhibitor group, however, no significant changes in mimics group. In conclusion, miR451 should be a tumor-suppressing gene in bladder cancer. miR-451 could maintain the bladder tumor cells in epithelial phenotype, inhibit EMT process, thereby reducing the invasion and migration of tumor cells.     

lncRNA HOXA11-AS对膀胱尿路上皮癌细胞增殖、迁移、侵袭的影响及其机制研究

目的:研究lncRNA HOXA11-反义RNA(HOXA11-AS)对膀胱尿路上皮癌J82细胞增殖、侵袭迁移的影响及与微小RNA-515-5p(miR-515-5p)之间的调控关系.方法:qRT-PCR检测膀胱尿路上皮癌组织、癌旁组织和J82细胞中HOXA11-AS和miR-515-5p的转录水平,MTT法和Tran...     

miR-411-5p对胃癌细胞凋亡、增殖和干样特性以及裸鼠成瘤的影响

目的 miR-411-5p对胃癌SGC-7901细胞生物学特性及裸鼠成瘤的影响.方法 荧光定量PCR(RT-qPCR)检测转染效率.5-溴脱氧尿嘧啶核苷(BrdU)染色检测增殖.流式细胞术检测凋亡.干细胞成球实验检测细胞干样特性.蛋白印迹实验检测增殖细胞核抗原(PCNA)、凋亡抑制蛋白(Survivin)、半胱天冬酶-...     

miR-218 调控SNX4 蛋白对乳腺癌细胞增殖和侵袭的影响

目的：探讨miR-218调控SNX4蛋白对乳腺癌细胞增殖和侵袭能力的影响。 方法：qPCR检测乳腺癌组织和乳腺癌细胞株中miR-218的表达情况;分析miR-218的表达和乳腺癌临床病理参数之间的关系;双荧光素酶实验检测miR-218和SNX4之间的关系;过表达miR-218后MTT实验和侵袭实验检测乳腺癌细胞增殖和侵袭能力变化;过表达SNX4后MTT实验和侵袭实验检测SNX4对乳腺癌细胞增殖和侵袭能力的恢复水平;裸鼠体外成瘤实验检测miR-218对乳腺癌细胞株成瘤能力的影响。结果：miR-218在乳腺癌组织和MCF-7细胞株中表达水平较高,miR-218的表达与乳腺癌的病理分期相关以及淋巴结转移情况有关, SNX4可能是miR-218下游的作用靶点;过表达miR-21可以抑制乳腺癌细胞的增殖侵袭能力,过表达SNX4后可以逆转miR-218对乳腺癌细胞的抑制作用,过表达miR-218后可以抑制乳腺癌细胞株在裸鼠体内的成瘤能力。结论：miR-218在乳腺癌中表达上调,同时miR-218可以调控SNX4的表达而影响乳腺癌细胞增殖和侵袭能力。     

MicroRNA-574-5p directly targets FOXN3 to mediate thyroid cancer progression via Wnt/β-catenin signaling pathway

ObjectiveThyroid cancer is the most common endocrine tumor. A large number of thyroid cancer-related miRNAs have been studied and identified. However, the detailed roles of miR-574-5p in thyroid cancer remain poorly understood. This work mainly aimed to investigate the role of miR-574-5p/FOXN3 axis and its mechanism in the thyroid cancer progression.MethodsMiR-574-5p, FOXN3, Wnt/β-catenin pathway, and apoptosis-related markers were measured by quantitative real-time PCR (qRT-PCR) and western blotting analysis, respectively. Luciferase reporter assay was employed to validate the direct targeting of FOXN3 by miR-574-5p. MTT, flow cytometry, wound healing and transwell experiments were applied to analyze the functions of FOXN3 and miR-574-5p in thyroid cancer cells.ResultsKnockdown of miR-574-5p up-regulated FOXN3 expression and miR-574-5p directly targeted FOXN3 in thyroid cancer cells. Biological function experiments showed that knockdown of miR-574-5p inhibited proliferation, migration, invasion and promoted apoptosis of thyroid cancer cells. The activation of Wnt/β-catenin pathway was suppressed by MiR-574-5p silencing. FOXN3 silencing reversed the effects of miR-574-5p inhibitor on FOXN3 level and Wnt/β-catenin singling pathway, also reversed the effects on cell migration, proliferation, invasion and apoptosis.ConclusionThe miR-574-5p/FOXN3 axis is a novel molecular mechanism that promotes thyroid cancer progression, suggesting their potential for clinical therapy of thyroid cancer.     

Adsorption of miR-218 by lncRNA HOTAIR regulates PDE7A and affects glioma cell proliferation,invasion, and apoptosis

Objective: To evaluate the role of targeted adsorption of miR-218 by long-chain non-coding RNAHOTAIR to regulate PDE7A on glioma cell proliferation, invasion, and apoptosis. Methods: The expressions of lncRNA HOTAIR, miR-218, and PDE7A in glioma tissues and normal parcancer tissues, NHA and glioma cell lines were determined, and correlations among the three genes were analyzed. The subcellular localization of lncRNA HOTAIR was determined by fluorescent in situ hybridization. Dual-luciferase reporter assay was used to validate the targeted relationship between lncRNA HOTAIR/miR-218/PDE7A. Glioma cells were grouped to receive intervention of lncRNA HOTAIR or miR-218. MTT, transwell, and flow cytometry were performed to determine the proliferation, invasion, and apoptosis of cells. Results: Compared with the normal tissues and cells, the expression of lncRNA HOTAIR was increased while miR-218 was suppressed in glioma tissues samples and cells (all P<0.05). Inhibition of lncRNA HOTAIR expression, was able to induce apoptosis and suppress the proliferation and invasion of cells (all P<0.05). LncRNA HOTAIR is mainly localized in the cytoplasm, and is able to adsorb miR-218 as ceRNA. The effect of knockdown of HOTAIR on glioma cells could be partially rescued by miR-218 inhibitor. The expression of PDE7A was enhanced in glioma tissues and cells compared to normal tissues and cells (all P<0.05), which positively correlated with the expression of HOTAIR (r=0.546, P<0.05) and negatively correlated with the expression of miR-218 (r=0.363, P<0.05). The targeted relationship between miR-218 and PDE7A was validated: Overexpression of miR-218 was able to suppress the proliferation and invasion of glioma cells and restrain apoptosis compared to the miR-NC group (all P<0.05). The effect of miR-218 on glioma cells could be partially rescued by PDE7A. Conclusion: lncRNA HOTAIR can adsorb miR-218 to regulate expression of PDE7A and promote the malignant biologic behavior of glioma cells.     

miR-766-5p 过表达介导 MMP-7 对胶质瘤细胞增殖、 侵袭与 凋亡的影响及机制

目的 探讨 miR-766-5p 对胶质瘤细胞增殖、 侵袭与凋亡的影响。 方法 Targetscan 预测 miR-766- 5p 与基质金属蛋白酶-7 (MMP-7) 靶向关系并进行验证, 构建 miR-766-5p 过表达、 MMP-7 过表达和 miR766-5p + MMP-7 过表达胶质瘤 U87 细胞系, BrdU 染色、 流式细胞法、 Transwell 实验、 Western 印迹分别检 测细胞增殖、 凋亡、 侵袭及相关蛋白表达。 结果 miR-766-5p 可靶向抑制 MMP-7 表达, miR-766-5p 过表达 可抑制 Ki67、 增殖细胞核抗原 (PCNA)、 Wnt1、 β-链蛋白 (β-catenin) 蛋白表达以抑制 U87 细胞增殖和侵 袭, 可促进 Bcl 相关 X 蛋白 (Bax) / B 细胞淋巴瘤/ 白血病-2 (Bcl-2) 蛋白表达以促进细胞凋亡, MMP-7 过表达可以缓解 miR-766-5p 过表达所致 U87 细胞增殖、 侵袭抑制作用和凋亡促进作用。 结论 miR-766-5p 可靶向抑制 MMP-7 表达抑制 U87 细胞胞增殖、 侵袭和促细胞凋亡。     

miR-454-3p靶向BPTF表达调控肺癌细胞A549增殖、迁移和侵袭

目的探讨miR-454-3p对肺癌细胞增殖、迁移和侵袭的影响及分子机制。方法采用RT-PCR技术检测miR-454-3p在肺癌组织以及肺癌细胞株中的表达,以表达量最低的肺癌细胞A549为后续分析对象,将miR-454-3p mimic转入A549细胞,RT-PCR验证miR-454-3p的过表达效率;采用CCK-8、迁移、侵袭等实验,观察转染组与对照组肺癌细胞的增殖、迁移和侵袭情况;生物信息学预测BPTF是miR-454-3p的靶标,构建BPTF 3′UTR荧光素酶载体,通过双荧光素酶报告基因验证miR-454-3p和BPTF的靶向关系;应用Western blot法检测BPTF的表达及迁移、侵袭相关蛋白的变化。结果RT-PCR实验表明miR-454-3p在肺癌组织和细胞中表达下调,且在A549细胞中表达最低(P<0.05)。与对照组相比,过表达miR-454-3p的肺癌细胞A549增殖能力明显降低,迁移和侵袭能力均受到抑制(P<0.05)。生物信息学软件分析BPTF为miR-454-3p的潜在靶基因,过表达miR-454-3p后BPTF的表达水平明显受到抑制。同时,迁移相关蛋白MMP-2和MMP-9表达明显下调,E-cadhern表达明显上调,而E-cadherin的负性调控N-cadherin表达下降。结论miR-454-3p可能通过靶向下调BPTF的表达,抑制肺癌细胞的增殖、迁移和侵袭能力,为肺癌的靶向治疗提供潜在靶标。     

CircRNA has_circ_0001806 promotes hepatocellular carcinoma progression via the miR-193a-5p/MMP16 pathway

Reportedly, circular RNAs (circRNAs) are crucial regulators in cancer progression. Nonetheless, the molecular mechanism of circRNAs in hepatocellular carcinoma (HCC) has not been fully clarified. Gene expression omnibus (GEO) database was employed to screen out the differentially expressed circRNAs in HCC. qRT-PCR and western blot were executed to detect circ_0001806 expression, miR-193a-5p expression, and MMP16 mRNA and protein expressions in HCC. The effect of circ_0001806 on HCC was analyzed by the CCK-8 method and Transwell experiment. RIP assay, pull-down experiment, and dual-luciferase reporter gene experiment were applied to validate the targeting relationships among circ_0001806, miR-193a-5p, and MMP16. Circ_0001806 was up-modulated in HCC tissues and cell lines. Knockdown of circ_0001806 impeded the multiplication, migration, and invasion of HCC cells. Circ_0001806 could up-regulate MMP16 expression through repressing miR-193a-5p, thereby facilitating the malignant biological behaviors of HCC. Circ_0001806 promoted HCC progression by regulating miR-193a-5p/MMP16 axis.     

miR-210-3p通过靶向ATG7抑制人膀胱癌细胞J82的自噬和诱导凋亡

目的探索miR-210-3p抑制人膀胱癌细胞J82机制。方法应用定量实时PCR(qRTPCR)检测miR-210-3p及其靶基因自噬相关基因7(autophagy-related gene 7,ATG7)在J82细胞中的表达。为验证miR-210-3p和ATG7之间的生物学关系,进行荧光素酶报告检测。通过体外实验研究miR-210-3p和ATG7在J82细胞中的生物学功能,包括CCK-8法检测细胞增殖情况,hoechst染色检测细胞凋亡,western blot检测细胞自噬。结果 miR-210-3p表达明显下调ATG7表达水平,生物信息学预测和荧光素酶报告实验证明miR-210-3p抑制ATG7是通过直接结合到ATG7 3’-未翻译区域(3’-UTR)。在J82细胞中,miR-210-3p上调可抑制ATG7表达、细胞自噬和细胞增殖,同时诱导细胞凋亡。结论 miR-210-3p通过负调控ATG7抑制细胞增殖和自噬,并促进凋亡,从而促进膀胱癌细胞J82细胞死亡。因此,在膀胱癌中抑制自噬对于开发针对膀胱癌的自噬靶向治疗至关重要。本研究补充了miRNA调控膀胱癌的相关机制。