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.     

Effect of estradiol and bisphenol A on human hepatoblastoma cell viability and telomerase activity

Sex hormones from environmental and physiological sources might play a major role in the pathogenesis of hepatoblastoma in children. This study investigated the effects of estradiol and bisphenol A on the proliferation and telomerase activity of human hepatoblastoma HepG2 cells. The cells were divided into 6 treatment groups: control, bisphenol A, estradiol, anti-estrogen ICI 182,780 (hereinafter ICI), bisphenol A+ICI, and estradiol+ICI. Cell proliferation was measured based on average absorbance using the Cell Counting-8 assay. The cell cycle distribution and apoptotic index were determined by flow cytometry. Telomerase activity was detected by polymerase chain reaction and a telomeric repeat amplification protocol assay. A higher cell density was observed in bisphenol A (P<0.01) and estradiol (P<0.05) groups compared with the control group. Cell numbers in S and G2/M phases after treatment for 48 h were higher (P<0.05), while the apoptotic index was lower (P<0.05) and telomerase activities at 48 and 72 h (P<0.05) were higher in these groups than in the control group. The cell density was also higher in bisphenol A+ICI (P<0.01) and estradiol+ICI (P<0.05) groups compared with the ICI group. Furthermore, cell numbers were increased in S and G2/M phases (P<0.05), while the apoptotic index was lower (P<0.05) and telomerase activities at 48 and 72 h were higher (P<0.05) in these groups than in the ICI group. Therefore, bisphenol A and estradiol promote HepG2 cell proliferation in vitro by inhibition of apoptosis and stimulation of telomerase activity via an estrogen receptor-dependent pathway.     

Increased expression of miR-21 predicts poor prognosis in patients with hepatocellular carcinoma

Background: MicroRNAs (miRNAs) play critical roles in hepatocellular carcinoma (HCC) development and progression. Aberrant miR-21 expression has been reported in several cancers. However, the clinical significance of miR-21 in human HCC is still unclear. Methods: A total of 112 patients with primary HCC who underwent a curative liver resection were included in this retrospective study. The differentially expressed amount of the miR-21 was validated by quantitative real-time PCR (qRT-PCR). Survival rate was analyzed by log-rank test, and survival curves were plotted according to Kaplan-Meier. Multivariate analysis of the prognostic factors was performed with Cox regression model. Results: As revealed by qRT-PCR analysis, miR-21 expression was significantly upregulated in HCC tissues when compared with adjacent non-tumor tissues (P<0.05). High miR-21 expression level was observed to be closely correlated with tumor differentiation, TNM stage and vein invasion (P<0.05). Patients who had high miR-21 expression had a shorter overall survival than patients who had low miR-21 expression (P<0.05). Moreover, multivariate analysis of the prognosis factors with a Cox proportional hazards model showed that high miR-21 expression was a significant independent predictor of poor survival in HCC (P<0.05). Conclusion: Our results suggested that increased expression of miR-21 was significantly correlated with tumor progression and could be a novel potential biomarker for HCC prognosis.     

miR-634 在肝癌中的表达及对肝癌细胞生物学行为的影响

目的:检测microRNA-634(miR-634)在肝癌中的表达水平及其对肝癌细胞常见生物学行为的调控作用。方法:采用实时荧光定量PCR(RT鄄qPCR)法检测肝癌细胞系(HepG2、SMMC7721、Bel7402、Bel7404、SNU739)、69 例肝癌组织及匹配癌旁组织中miR-634 的相对定量,分析miR-634 表达与肝癌患者性别、年龄、肿瘤直径、分化程度、Child-Pugh 分级、BCLC 分期、门静脉癌栓及肝外转移的关系,同时构建miR-634 的真核表达载体并转染肝癌细胞系,采用活细胞计数试剂盒CCK-8、流式细胞仪Annexin V/ PI 双染法和Transwell 侵袭实验检测转染miR鄄634 对细胞增殖、凋亡和侵袭能力的影响。结果:与正常人肝细胞系L-02 相比,肝癌细胞的miR-634 水平均降低(P <0.05),表达量依次为HepG2 >SNU739 >Bel7402 > Bel7404 >SMMC7721;69 例肝癌组织的miR鄄634 水平为(0.253±0.019),低于匹配癌旁组织(P<0.05),且与肿瘤直径、分化程度、BCLC分期、门静脉癌栓及肝外转移均有关(P<0.05)。过表达组转染24 ~96 h 后的miR鄄634 水平持续升高,与对照组和空转染组的差异有统计学意义(P<0.05);与对照组和空转染组相比,转染组的增殖抑制率、凋亡率均升高,但穿膜细胞数降低,差异有统计学意义(P<0.05)。结论:miR-634 在肝癌组织和细胞中均为低表达,且与临床病理参数有关,上调其水平可抑制肝癌细胞增殖及侵袭并诱导凋亡,对于肝癌防治有重要借鉴价值。     

Diagnostic significance of miR-106a in gastric cancer

As one of the most common malignant tumors, gastric cancer still lacks tumor markers with enough specificity and sensitivity. Therefore the development of novel tumor markers is necessary for early diagnosis in clinics. MicroRNA (miR) has been known to be of unique expressional patterns in various tumors and may work as potential tumor markers for clinical use. This study thus explored the significance of plasma miR-106a in clinical diagnosis of gastric cancer and its effects on proliferation of cancer cells. Plasma miR-106a levels were quantified by real-time quantitative fluorescent PCR methods in 80 cases of gastric cancer patients and healthy individuals to analyze the correlation between miR-106a and clinical features. MiR-106 inhibitor was further transfected into human gastric carcinoma cells for further cell proliferation using CCK-8 approach. MiR-106a was significantly up-regulated in gastric cancer patient plasma samples compared to healthy individuals (P<0.01). The area under ROC curve was 0.895 (95% CI: 0.846~0.943). It has a specificity of 93.8% and a sensitivity of 77.5% in diagnosing gastric cancer. MiR-106a level was also correlated with cancer differentiation stage, lymph node metastasis, TNM stage and tumor size (P<0.05). The down-regulation of miR-106 in gastric carcinoma cells inhibited cell proliferation (P<0.05). MiR-106a was significantly up-regulated in gastric cancer patients and can facilitate the in vitro proliferation of tumor cells. It may work as a biological marker for gastric cancer.     

MicroRNA-200a inhibits epithelial-mesenchymal transition in human hepatocellular carcinoma cell line

Objective: Our study investigated the role of microRNA (miR)-200a and its molecular targets in hepatocellular carcinoma (HCC) cells. Methods: An inhibitor of miR-200a was transiently transfected into the hepatocellular carcinoma cell line, MHCC-97L. The effect of this transfection on mRNA levels of epithelial-mesenchymal transition (EMT)-related genes was measured by fluorescence-based quantitative real-time polymerase chain reaction (qRT-PCR). Further, protein levels of EMT-related genes, cell proliferation and apoptosis-related markers were assessed by Western blot analysis in these transfected cells. MTT and wound-healing assay were used to evaluate the proliferation and migration of MHCC-97L cells in presence and in absence of miR-200a inhibitor. Results: Compared with miR-NC control group, qRT-PCR results in anti-miR-200a group revealed a significant reduction in the mRNA levels of E-cadherin, with a concomitant increasing in vimentin mRNA level (all P < 0.05). Western blot results showed higher E-cadherin and Caspase-3 protein expressions in anti-miR-200a group compared to miR-NC group (P < 0.05). In addition, vimentin and Ki-67 protein expression was found sharply decreased in anti-miR-200a group compared to miR-NC group (P < 0.05). Consistent with this, wound-healing and MTT assay showed that migration and proliferation capacity of MHCC-97L cells in anti-miR-200a group is significantly increased compared with miR-NC group (both P < 0.05). Conclusion: Our study reveals an important role of miR-200a in inhibiting EMT, proliferation and migration in HCC cells, suggesting the possibility of miR-200a-based therapeutics in HCC.     

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.     

Cyclic RNA Circ_0000735 sponges miR-502-5p to promote bladder cancer cell proliferation and invasion and inhibit apoptosis

Objective: The objective of this study was to investigate the effect on the proliferation, invasion, and apoptosis of bladder cancer cells through miR-502-5p of the Circ_0000735 circular RNA. Methods: Circ_0000735 and miR-502-5p expression of bladder cancer patients in malignant and paracancerous tissues was identified using qRT-PCR. Nucleoplasm isolation assay and RNase R enzymatic assay were used to classify Circ_0000735 subcellular origin and stability. Dual luciferase reporter assay and RIP assay were used to confirm Circ_0000735 and miR-502-5p targeting relationships. Cell proliferation, apoptosis, and invasion capacity were identified using CCK8, flow cytometry, and transwell assays. To confirm the effect of Circ_0000735 on tumorigenesis in nude mice, in vivo experiments were conducted. Results: Circ_0000735 expression was increased in bladder cancer tissues and cells compared with paraneoplastic tissues and normal cells, and miR-502-5p expression was reduced (both P<0.05). In the cytoplasm, Circ_0000735 was largely clustered and could not be digested by the RNase R enzyme, and ceRNA may play a role in bladder cancer cells. Circ_0000735 silencing prevented cell proliferation and invasion and facilitated apoptosis (all P<0.05). The incorporation of miR-502-5p inhibitor rescued the effect on bladder cancer cells of Circ_0000735 silencing. In vitro experiments showed that inhibition of Circ_0000735 expression was beneficial in suppressing tumorigenic ability in nude mice. Conclusion: Circ_0000735 can adsorb miR-502-5p to promote bladder cancer cell proliferation and invasion and inhibit apoptosis. Circ_0000735 may be an effective molecular target for bladder cancer therapy.     

miR-485-5p靶向作用于SOX5抑制肝癌细胞Hep3B的活力及侵袭和迁移能力

目的:研究微小RNA-485-5p(miR-485-5p)对肝癌细胞的活力及迁移和侵袭能力的影响及其潜在机制。方法:以正常肝细胞THLE-3为对照,采用RT-qPCR检测肝癌细胞中性别决定区Y框蛋白5(SOX5)mRNA和miR-485-5p的表达,Western blot检测SOX5、增殖细胞核抗原(PCNA)、Ki67、细胞周期蛋白D1(cyclin D1)及基质金属蛋白酶2(MMP-2)的表达水平,MTT法检测Hep3B细胞的活力,Transwell实验检测细胞的迁移和侵袭能力,双萤光素酶报告基因系统验证细胞中miR-485-5p与SOX5的调控关系。结果:与对照组相比,在肝癌细胞Hep3B、Huh7和HCCLM3中miR-485-5p的表达水平显著降低(P<0.05),SOX5的mRNA和蛋白表达水平均显著升高(P<0.05);过表达miR-485-5p可抑制Hep3B细胞的活力及迁移和侵袭能力(P<0.05);miR-485-5p靶向调控SOX5的表达,敲减SOX5的表达也可抑制Hep3B细胞的活力及迁移和侵袭能力;过表达SOX5可部分逆转过表达miR-485-5p对Hep3B细胞活力及迁移和侵袭能力的抑制作用(P<0.05)。结论:miR-485-5p通过靶向调控SOX5基因抑制Hep3B细胞的活力及迁移和侵袭能力,有望成为肝癌诊断和治疗的潜在分子靶点。     

MicroRNA-539靶向调控E2F转录因子3抑制肝癌的发生与发展

目的 探讨肝癌组织中microRNA-539（miR-539）表达水平及其抑制肝癌细胞增殖的作用机制。方法 收集90例肝癌患者术后肝癌组织及癌旁组织标本,用实时定量聚合酶链反应（Real-time PCR）检测组织中miR-539表达量。在PCL/PRF5、Huh7和QGY-7701肝癌细胞中转染miR-539 模拟物（mimics）,用Real-time PCR检测miR-539表达量。用MTT法检测miR-539对肝癌细胞增殖的影响;用流式细胞仪检测miR-539对肝癌细胞凋亡的影响;用免疫印迹法检测miR-539对肝癌细胞中E2F转录因子3（E2F3）蛋白表达的影响。结果 肝癌组织中miR-539表达量显著低于癌旁组织（P<0.05）;miR-539表达量与肝癌患者性别、年龄及肝癌组织分化程度无关（P>0.05）;与肿瘤直径和淋巴结转移相关（P<0.05）。转染miR-539 mimics组肝癌细胞中miR-539表达量显著高于空白组和miR-539 NC组（P<0.05）。在QGY-7701细胞中,过表达miR-539能显著抑制细胞增殖（P<0.05）,降低E2F3蛋白的表达（P<0.05）,对细胞凋亡无明显作用（P>0.05）。结论 miR-539是肝癌发生中的抑癌因子,可能是通过靶向下调E2F3蛋白水平达到抑制癌细胞的增殖。     

miR-21 inhibitor suppresses proliferation and migration of nasopharyngeal carcinoma cells through down-regulation of BCL2 expression

This study is to investigate the expression of miR-21 in nasopharyngeal carcinoma (NPC) cells, and the effect of miR-21 in the biological behavior and expression of B-cell lymphoma 2 (BCL2) in NPC cells. Paired NPC and adjacent non-tumor tissues were obtained from 53 patients who underwent primary surgical resection of NPC tissues. Luciferase reporter assay was performed to test whether BCL2 is a direct target of miR-21. Methylthiazolyl blue tetrazolium assay and colony assay were used to evaluate the effect of miR-21 on NPC cell proliferation. Transwell and wound-healing assays were carried out to test the effect of low expression of miR-21 on cancer cell migration and invasion. QRT-PCR and Western blotting were used to measure the levels of mRNA and protein expression, respectively. Tumor tissues showed a positive correlation between the levels of miR-21 and BCL2 protein expression. Cells transfected with miR-21 inhibitor healed slower compared the control (P < 0.05). In addition, cell migration was notably inhibited by the down-regulation of miR-21 in vitro (P < 0.05). The reduction in miR-21 expression showed a remarkable effect on the biological behavior of NPC cell clone formation (P < 0.05). Low expression of miR-21 by transfection with miRNA expression plasmid led to a decrease in BCL2 expression, which was accompanied by reduced migration and proliferation of the cancer cells. Our results demonstrated that miR-21 inhibitor down-regulated BCL2 expression level, suggesting that BCL2 might be a target gene for the initiation and development of NPC cells.     

miR-892a靶向调控SOX5对前列腺癌细胞增殖和迁移能力的影响

目的 探讨miR-892a对前列腺癌细胞的增殖和迁移能力的影响。方法 从癌症基因组图谱(TCGA)数据库下载499例前列腺肿瘤及52例癌旁组织的微小RNA(miRNA)的样本资料,再采用R语言的limma包及gplots包进行差异表达分析,绘制火山图和热图,筛选目的基因。采用荧光定量反转录聚合酶链式反应(RT-PCR)检测前列腺上皮细胞(RWPE-1)和前列腺癌细胞(DU145、PC3)中miR-892a表达情况。将miR-892a模拟物(miR-892a mimic组)及阴性对照(miR-NC组)分别转染前列腺癌PC3细胞,采用四甲基偶氮唑蓝(MTT)、低密度集落形成实验及Transwell小室细胞迁移实验检测两组细胞的增殖及迁移,通过生物信息学在线预测网站预测miR-892a的潜在靶基因为性别决定区Y框蛋白5(SOX5),RT-PCR、Western blot实验及荧光素酶报告基因实验验证miR-892a靶向调控SOX5。结果 筛选的目的基因为miR-892a。前列腺癌细胞PC3和DU145中miR-892a的相对表达量分别为0.41±0.40和0.47±0.45,明显低于前列腺上皮细胞的1.18±0.35,差异有统计学意义(F=340.330, P＜0.01),选择PC3细胞作为后续研究对象。MTT增殖实验结果显示,miR-892a mimic组在24 h、48 h、72 h和96 h 光度值均低于miR-NC组,差异均有统计学意义(P值均＜0.05)。低密度细胞集落形成实验中,过表达miR-892a后,miR-892a minic组细胞克隆数为251.00±64.84,miR-NC组为433.28±62.93;Transwell小室细胞迁移实验中miR-892a mimic组穿过小室的细胞数为340.33±31.30,miR-NC组为534.33±61.07,差异均有统计学意义(t=-3.194、-4.906,P值均＜0.05)。miR-NC组和miR-892a mimic组的SOX5 mRNA相对表达水平分别为1.021±0.012和0.156±0.017,SOX5的蛋白表达水平分别为0.519±0.014和0.196±0.019,两组差异均有统计学意义(t=71.807、24.097, P值均＜0.01)。野生型miR-892a mimic组荧光素酶比值为0.37±0.08,miR-NC组为0.97±0.10,差异有统计学意义(t=12.890, P＜0.01)。SOX5的蛋白表达水平低于于NC组(t=71.807, P＜0.01),并且miR-892a能够直接靶向SOX5发挥生物学效应(t=12.890, P＜0.01)。结论 与前列腺上皮细胞相比较,miR-892a在前列腺癌PC3细胞中显著低表达;过表达miR-892a能够抑制前列腺癌细胞的增殖和迁移,并靶向调控SOX5。     

Role of MAPK7 in cell proliferation and metastasis in ovarian cancer

Aim: To investigate the effect of mitogen-activated protein kinase 7 (MAPK7) in ovarian cancer metastasis and to explore its potential mechanism. Methods: pcDNA-MAPK7 and siRNA-MAPK7 vectors were transfected into the human ovarian cell line OVCAR-3 based on gene silencing and overexpression methods. Effects of MAPK7 overexpression and silencing on OVCAR-3 cells proliferation, cell invasion, and migration were analyzed using the MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide) assay, Matrigel methods, and Markered methods respectively. In addition, effect of MAPK7 expression on extracellular matrix (ECM) associated protein was detected using Western blot. Results: Compared with the controls, MAPK7 was up-regulated when cells were transfected with pcDNA-MAPK7 plasma, as well as MAPK7 was sliced when cells were transfected with siRNA-MAPK7 plasma (P<0.05). Besides, biological function analysis performed that overexpression of MAPK7 significantly increased OVCAR-3 cell proliferation, invasion, and migration (P<0.05), while these effects were inhibited by MAPK7 silencing (P<0.05). Additionally, MAPK7 overexpression increased type II collagen expression (P<0.05). However, there was no significant difference between MAPK7 expression and type I collagen expression (P>0.05). Conclusion: Our data implied the up-regulated MAPK7 might contribute to ovarian cancer metastasis through up-regulating type II collagen expression and then were involved in cell biological processes such as cell proliferation, invasion, and migration. MAPK7 may be a potential therapeutic target in the clinical treatment for ovarian cancer.     

Bevacizumab induces A549 cell apoptosis through the mechanism of endoplasmic reticulum stress in vitro

Aims: To observe the effect of bevacizumab on human A549 cells and explore its mechanism. Methods: After different concentrations (0 μM, 1 μM, 5 μM, 25 μM) of bevacizumab treating in A549 cells, CCK8 assay detect the impact of bevacizumab on A549 cell proliferation and flow cytometry determine the effect of bevacizumab on human A549 cells apoptosis. Real-time PCR and Western blotting detect the changing expression of the target gene (CHOP, caspase-4, IRE1, XBP-1) on mRNA and Protein level. Results: Treatment with bevacizumab for 24-hr have induced cell death in a does-dependent manner dramatically (P<0.05). In terms of the mRNA level, expression of XBP-1 has increased obviously in each group (1 μM, 5 μM, 25 μM) (P<0.01); the expression of CHOP (25 μM) and caspase-4 (5 μM) have increased slightly (P<0.05). In terms of the protein level, the expression of CHOP has increased obviously in each group (1 μM, 5 μM, 25 μM) when compared with the control group (0 μM) (P<0.05). As for caspase-4 (5 μM, 25 μM), the expression have increased slightly when compared with the control group (0 μM) (P<0.05). Conclusion: Bevacizumab can induce A549 cell apoptosis through the mechanism of endoplasmic reticulum stress.     

miR-125b 调控ART4 蛋白对肝癌细胞增殖和侵袭的影响

目的:探讨miR-125b 对ART4 蛋白的调控作用及在肝癌细胞中对其增殖和侵袭能力的影响。方法:qPCR 检测肝癌组织和肝癌细胞中miR-125b 的表达情况;过表达miR-125b 后qPCR 检测ART4 的表达;双荧光素酶实验检测miR-125b和ART4 的关系;MTT 实验检测过表达miR-125b 对肝癌细胞增殖能力的影响;Transwell 侵袭实验检测miR-125b 的过表达对肝癌细胞侵袭能力的影响;MTT 实验检测过表达ART4 后逆转miR-125b 对肝癌细胞增殖能力的影响;Transwell 侵袭实验检测过表达ART4 后逆转miR-125b 对肝癌细胞侵袭能力的影响。结果:miR-125b 在肝癌细胞中表达水平较低,过表达miR-125b可以有效抑制ART4 蛋白的表达;过表达miR-125b 可以抑制肝癌细胞的增殖侵袭能力;过表达ART4 后可以逆转肝癌细胞被miR-125b 抑制的增殖侵袭能力。结论:miR-125b 在肝癌中表达下调,同时miR-125b 可以调控ART4 的表达影响肝癌细胞增殖和侵袭能力。     

Down-regulation of microRNA-126 and microRNA-133b acts as novel predictor biomarkers in progression and metastasis of non small cell lung cancer

Objective: MiRNAs play crucial roles in progression of cancer. However, the underlying mechanisms of miRNAs in non small cell lung cancer are still poorly understood. The aim of this study was to investigate the expression level of microRNA-126 (miR-126) and microRNA-133b (miR-133b) and also their association with clinicopathological features in patients with non small cell lung cancer (NSCLC). Methods: Total RNA was purified from NSCLC tissues and adjacent non-tumor tissues and then quantitative real-time PCR (qRT-PCR) was used to evaluate the expression rate of microRNAs. Furthermore, the association of miR-126 and miR-133b level with clinicopathological features and prognosis were evaluated. Results: Our findings showed that expression of miR-126 was decreased in NSCLC tissues compared with adjacent non-tumor tissues. On the other hand, a lower expression of miR-133b was seen in NSCLC tissues when compared with adjacent non-tumor tissues. In term of miR-126, our results showed that miR-126 was associated with tumor stage and lymph nodes metastasis (P<0.05). In term of miR-133b, our finding indicated that decreased expression of miR-133b was correlated with advanced tumor stage and lymph nodes metastasis (P<0.05). Kaplan-Meier analysis and log-rank test indicated that patients with low expression of miR-126 and miR-133b had a shorter overall survival (log-rank test; P<0.05). Multivariate Cox proportional hazards model revealed that low expression of miR-126 and miR-133b, advanced tumor stage and lymph nodes metastasis were independent prognostic factors for overall survival of NSCLC patients. Conclusions: These findings suggested that miR-126 and miR-133b might play a key role in the progression and metastasis of NSCLC and would be applied as a novel therapeutic agent.     

Rat visceral yolk sac cells: viability and expression of cell markers during maternal diabetes

The function of the visceral yolk sac (VYS) is critical for embryo organogenesis until final fetal development in rats, and can be affected by conditions such as diabetes. In view of the importance of diabetes during pregnancy for maternal and neonatal health, the objective of this study was to assess fetal weight, VYS cell markers, and viability in female Wistar rats (200-250 g) with induced diabetes (alloxan, 37 mg/kg) on the 8th gestational day (gd 8). At gd 15, rats from control (n=5) and diabetic (n=5) groups were anesthetized and laparotomized to remove the uterine horns for weighing of fetuses and collecting the VYS. Flow cytometry was used for characterizing VYS cells, and for determining mitochondrial activity, cell proliferation, DNA ploidy, cell cycle phases, and caspase-3 activity. Fetal weight was reduced in the diabetic group. Expression of the cell markers CD34, VEGFR1, CD115, CD117, CD14, CCR2, CD90, CD44, STRO-1, OCT3/4, and Nanog was detected in VYS cells in both groups. In the diabetic group, significantly decreased expression of CD34 (P<0.05), CCR2 (P<0.001), and OCT3/4 (P<0.01), and significantly increased expression of CD90 (P<0.05), CD117 (P<0.01), and CD14 (P<0.05) were observed. VYS cells with inactive mitochondria, activated caspase-3, and low proliferation were present in the rats with diabetes. Severe hyperglycemia caused by maternal diabetes had negative effects on pregnancy, VYS cell viability, and the expression of cell markers.     

Overexpression of miR-221 inhibits proliferation and promotes apoptosis of human astrocytoma cells

microRNAs (miRNAs) play tumor-promoting roles in a variety of tumors. This study investigated the expression of miRNA-211 (miR-221) in human astrocytoma, and its effect on proliferation and apoptosis of human astrocytoma cells in vitro. miR-221 expression was detected in 10 astrocytoma tissues and 4 adjacent tissues by real-time quantitative PCR (qRT-PCR). miR-221 expression in situ was significantly higher in astrocytoma tissues than in adjacent tissues (P<0.05). To determine whether the upregulation of miR-221 could be associated with tumor development or progression, a synthetic miR-221 mimic was transiently transfected into U251 astrocytoma cells in vitro. qRT-PCR confirmed that the mimic significantly increased the expression of miR-221 in these cells. An MTT colorimetric assay indicated that proliferation was significantly higher in U251 cells transfected with miR-221 mimic than in scramble-transfected control cells (P<0.05). Further analysis of miR-221 transfected cells by flow cytometry revealed an altered cell cycle progression, with more cells in S and G1 phase, as well as an inhibition of apoptosis (P<0.05). These findings indicate that the upregulation of miR-221 in astrocytoma tissues may be associated with development or progression of these tumors. Thus, miR-221 should be explored as a potential molecular marker for the diagnosis and treatment of astrocytoma.