miR-599 Inhibits Vascular Smooth Muscle Cells Proliferation and Migration by Targeting TGFB2

Aberrant proliferation and migration of vascular smooth muscle cells (VSMCs) play a crucial role in the pathogenesis of cardiovascular diseases including coronary heart disease, restenosis and atherosclerosis. MicroRNAs are a class of small, non-coding and endogenous RNAs that play critical roles in VSMCs function. In this study, we showed that PDGF-bb, as a stimulant, promoted VSMCs proliferation and suppressed the expression of miR-599. Moreover, overexpression of miR-599 inhibited VSMCs proliferation and also suppressed the PCNA and ki-67 expression. In addition, we demonstrated that ectopic expression of miR-599 repressed the VSMCs migration. We also showed that miR-599 inhibited type I collagen, type V collagen and proteoglycan expression. Furthermore, we identified TGFb2 as a direct target gene of miR-599 in VSMCs. Overexpression of TGFb2 reversed miR-599-induced inhibition of VSMCs proliferation and type I collagen, type V collagen and proteoglycan expression. In conclusion, our findings suggest miR-599 plays a crucial role in controlling VSMCs proliferation and matrix gene expression by regulating TGFb2 expression.     

miR-455 Inhibits HepG2 Cell Proliferation and Promotes Apoptosis by Targeting RhoC

Molecular Biology - Hepatocellular carcinoma (HCC) is a common malignancy worldwide with poor prognosis and high mortality. The aberrant expression or alteration of microRNAs (miRNAs) contributes...     

沉默胃泌素基因抑制胃癌细胞的增殖、迁移和侵袭

胃癌细胞分泌的胃泌素与胃癌的发生、发展密切相关.为了探讨胃泌素对胃癌细胞增殖、迁移和侵袭的影响,本文构建靶向胃泌素基因的siRNA表达载体, 转染胃癌细胞AGS, 成功获得沉默胃泌素基因的稳转胃癌细胞株AGS/Gas-siRNA. 用MTT实验、软琼脂集落形成实验、细胞伤愈实验、Transwell实验及ELISA检测沉默胃泌素基因后细胞的增殖、迁移、侵袭及转移相关蛋白基质金属蛋白酶-2（MMP-2）和血管内皮生长因子（VEGF）的含量. 结果显示: 与空载体转染的对照细胞比较, 沉默胃泌素基因的细胞, 其增殖率和克隆形成率显著降低,迁移和侵袭到Transwell下室的细胞数分别降低了31.6 %和34 %. 培养上清液中MMP-2和VEGF含量也低于对照细胞. 结果提示,沉默胃泌素基因的胃癌细胞,通过降低MMP 2和VEGF分泌,抑制了细胞的增殖、迁移和侵袭, 这可能是胃泌素促进胃癌侵袭转移的机制之一.     

MiR-210-3p Inhibits Proliferation and Migration of C6 Cells by Targeting Iscu

Neurochemical Research - Glioma is the most common primary brain tumor and the most malignant type of glioma is glioblastoma with the character of high mortality, high recurrence rate and poor...     

Down-Regulation of Gab1 Inhibits Cell Proliferation and Migration in Hilar Cholangiocarcinoma

Hilar cholangiocarcinoma is a highly aggressive malignancy originating from the hilar biliary duct epithelium. Due to few effective comprehensive treatments, the prognosis of hilar cholangiocarcinoma is poor. In this study, immunohistochemistry was first used to detect and analyze the expression of Gab1, VEGFR-2, and MMP-9 in hilar cholangiocarcinoma solid tumors and the relationships to the clinical pathological features. Furthermore, Gab1 and VEGFR-2 siRNA were used to interfere the hilar cholangiocarcinoma cell line ICBD-1 and then detect the PI3K/Akt signaling pathway, MMP-9 levels and malignant biological behaviors of tumor cells. The data showed that 1. Gab1, VEGFR-2, and MMP-9 were highly expressed and positively correlated with each other in hilar cholangiocarcinoma tissues, which were related to lymph node metastasis and differentiation. 2. After Gab1 or VEGFR-2 siRNA interference, PI3K/Akt pathway activity and MMP-9 levels were decreased in ICBD-1 cells. At the same time, cell proliferation decreased, cell cycle arrested in G1 phase, apoptosis increased and invasion decreased. These results suggest that the expression of Gab1, VEGFR-2, and MMP-9 are significantly related to the malignant biological behavior of hilar cholangiocarcinoma. Gab1 regulates growth, apoptosis and invasion through the VEGFR-2/Gab1/PI3K/Akt signaling pathway in hilar cholangiocarcinoma cells and influences the invasion of tumor cells via MMP-9.     

MicroRNA-135a Inhibits Cell Proliferation by Targeting Bmi1 in Pancreatic Ductal Adenocarcinoma

Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal solid tumor due to the lack of reliable early detection markers and effective therapies. MicroRNAs (miRNAs), noncoding RNAs that regulate gene expression, are involved in tumorigenesis and have a remarkable potential for the diagnosis and treatment of malignancy. In this study, we investigated aberrantly expressed miRNAs involved in PDAC by comparing miRNA expression profiles in PDAC cell lines with a normal pancreas cell line and found that miR-135a was significantly down-regulated in the PDAC cell lines. The microarray results were validated by qRT-PCR in PDAC tissues, paired adjacent normal pancreatic tissues, PDAC cell lines, and a normal pancreas cell line. We then defined the tumor-suppressing significance and function of miR-135a by constructing a lentiviral vector to express miR-135a. The overexpression of miR-135a in PDAC cells decreased cell proliferation and clonogenicity and also induced G1 arrest and apoptosis. We predicted Bmi1 may be a target of miR-135a using bioinformatics tools and found that Bmi1 expression was markedly up-regulated in PDAC. Its expression was inversely correlated with miR-135a expression in PDAC. Furthermore, a luciferase activity assay revealed that miR-135a could directly target the 3''-untranslated region (3''-UTR) of Bmi1. Taken together, these results demonstrate that miR-135a targets Bmi1 in PDAC and functions as a tumor suppressor. miR-135a may offer a new perspective for the development of effective miRNA-based therapy for PDAC.     

MiR-122 Inhibits Cell Proliferation and Tumorigenesis of Breast Cancer by Targeting IGF1R

miRNAs are emerging as critical regulators in carcinogenesis and tumor progression. Recently, microRNA-122 (miR-122) has been proved to play an important role in hepatocellular carcinoma, but its functions in the context of breast cancer (BC) remain unknown. In this study, we report that miR-122 is commonly downregulated in BC specimens and BC cell lines with important functional consequences. Overexpression of miR-122 not only dramatically suppressed cell proliferation, colony formation by inducing G1-phase cell-cycle arrest in vitro, but also reduced tumorigenicity in vivo. We then screened and identified a novel miR-122 target, insulin-like growth factor 1 receptor (IGF1R), and it was further confirmed by luciferase assay. Overexpression of miR-122 would specifically and markedly reduce its expression. Similar to the restoring miR-122 expression, IGF1R downregulation suppressed cell growth and cell-cycle progression, whereas IGF1R overexpression rescued the suppressive effect of miR-122. To identify the mechanisms, we investigated the Akt/mTOR/p70S6K pathway and found that the expression of Akt, mTOR and p70S6K were suppressed, whereas re-expression of IGF1R which did not contain the 3′UTR totally reversed the inhibition of Akt/mTOR/p70S6K signal pathway profile. We also identified a novel, putative miR-122 target gene, PI3CG, a member of PI3K family, which further suggests miR-122 may be a key regulator of the PI3K/Akt pathway. In clinical specimens, IGF1R was widely overexpressed and its mRNA levels were inversely correlated with miR-122 expression. Taken together, our results demonstrate that miR-122 functions as a tumor suppressor and plays an important role in inhibiting the tumorigenesis through targeting IGF1R and regulating PI3K/Akt/mTOR/p70S6K pathway. Given these, miR-122 may serve as a novel therapeutic or diagnostic/prognostic-target for treating BC.     

miR-150-5p Inhibits Hepatoma Cell Migration and Invasion by Targeting MMP14

Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related mortality worldwide. Despite progress in diagnostics and treatment of HCC, its prognosis remains poor because the molecular mechanisms underlying hepatocarcinogenesis are not well understood. In the study, we focused on identifying the role of miRNAs in HCC progression. miRNA microarray was used to analyze the differentially expressed miRNAs, and the results were validated by qPCR. We found that the miR-150-5p expression is down-regulated in HCC tissues compared with pair non-tumor tissues. miR-150-5p expression is also decreased in metastatic cancer tissues compared with pair primary tissues, indicating that miR-150-5p may be involved in HCC metastasis. Functionally, miR-150-5p inhibition significantly promotes hepatoma cell migration and invasion, whereas miR-150-5p overexpression suppresses cancer cell migration and invasion in vitro. The matrix metalloproteinase 14 (MMP14) is identified as a new target gene of miR-150-5p. miR-150-5p markedly inhibits MMP14 expression in hepatoma cells, and miR-150-5p expression is negative correlation with MMP14 expression in vivo. More important, re-expression of MMP14 in hepatoma cells partially reverses the effect of miR-150-5p in inhibiting cell invasion.     

Nordentatin Inhibits Neuroblastoma Cell Proliferation and Migration through Regulation of GSK-3 Pathway

Cancer is caused by abnormal cell changes leading to uncontrolled cell growth. The specific characteristics of cancer cells, including the loss of apoptotic control and the ability to migrate into and invade the surrounding tissue, result in cancer cell metastasis to other parts of the body. Therefore, the inhibition of the proliferation, migration, and invasion of cancer cells are the principal goals in the treatment of cancer. This study aimed to investigate the inhibitory activity of nordentatin, a coumarin derivative isolated from Clausena harmandiana, regarding the proliferation and migration of human neuroblastoma cells (SH-SY5Y). Nordentatin at a concentration of 100 µM showed cell cytotoxicity toward SH-SY5Y that was significantly different from that of the control group (p < 0.01) at 24, 48, and 72 h. Moreover, nordentatin inhibited SH-SY5Y proliferation by inhibiting the antiapoptotic protein Mcl-1, leading to the cleavage of caspase-3 and resulting in the inhibition of a migratory protein, MMP-9, through the GSK-3 pathway (compared with cells treated with a GSK inhibitor). These results suggest that nordentatin inhibited the proliferation and migration of neuroblastoma cells through the GSK-3 pathway.     

Ikaros通过靶向调控 c-KIT抑制B-ALL白血病细胞的增殖

Ikaros是一种重要的造血细胞分化与发育调控因子,其基因结构、蛋白质活性的改变与淋巴细胞白血病的发生密切相关。致癌基因c-KIT与白血病的发生有直接联系,但Ikaros与c-KIT之间的调控关系尚未见报道。本研究报道,在人急性B淋巴细胞白血病（B-acute lymphoblastic leukemia, B-ALL）细胞中,Ikaros可靶向调控c KIT基因的转录与蛋白质表达。通过在人B ALL细胞系Nalm6中分别高表达和shRNA干扰Ikaros后,qRT-PCR与 Western 印迹结果显示,Ikaros可直接抑制c-KIT基因的表达。双荧光素酶报告实验检测Ikaros及其突变体对c-KIT基因的直接靶向作用。结果显示,野生型Ikaros可明显抑制c-KIT的表达,而突变体则不能。进一步利用染色质免疫共沉淀技术（chromatin immunoprecipitation,ChIP）,检测Ikaros对c-KIT上游启动子序列的结合活力。结果显示,Ikaros蛋白在c KIT的上游调控区约-500 bp处有明显的结合。Ikaros通过靶向c-KIT上游启动子,抑制c-KIT表达,抑制B-ALL细胞的增殖,为临床治疗白血病提供了新思路。     

Bindarit Inhibits Human Coronary Artery Smooth Muscle Cell Proliferation,Migration and Phenotypic Switching

Bindarit, a selective inhibitor of monocyte chemotactic proteins (MCPs) synthesis, reduces neointimal formation in animal models of vascular injury and recently has been shown to inhibit in-stent late loss in a placebo-controlled phase II clinical trial. However, the mechanisms underlying the efficacy of bindarit in controlling neointimal formation/restenosis have not been fully elucidated. Therefore, we investigated the effect of bindarit on human coronary smooth muscle cells activation, drawing attention to the phenotypic modulation process, focusing on contractile proteins expression as well as proliferation and migration. The expression of contractile proteins was evaluated by western blot analysis on cultured human coronary smooth muscle cells stimulated with TNF-α (30 ng/mL) or fetal bovine serum (5%). Bindarit (100–300 µM) reduced the embryonic form of smooth muscle myosin heavy chain while increased smooth muscle α-actin and calponin in both TNF-α- and fetal bovine serum-stimulated cells. These effects were associated with the inhibition of human coronary smooth muscle cell proliferation/migration and both MCP-1 and MCP-3 production. The effect of bindarit on smooth muscle cells phenotypic switching was confirmed in vivo in the rat balloon angioplasty model. Bindarit (200 mg/Kg/day) significantly reduced the expression of the embryonic form of smooth muscle myosin heavy chain, and increased smooth muscle α-actin and calponin in the rat carodid arteries subjected to endothelial denudation. Our results demonstrate that bindarit induces the differentiated state of human coronary smooth muscle cells, suggesting a novel underlying mechanisms by which this drug inhibits neointimal formation.     

新霉胺通过抑制血管生成素活性抑制人黑色素瘤细胞增殖、迁移和侵润

新霉胺（neamine）是一种无毒的新霉素（neomycin）降解产物;已有研究证明,其可抑制血管生成素（angiogenin,ANG）诱导的内皮细胞血管生成作用,阻滞异种移植的人结肠腺癌在裸鼠的生长.本研究证明,新霉胺对人黑色瘤细胞株A375的细胞增殖、迁移和侵润作用.MTT法及软琼脂培养显示,新霉胺可明显抑制A375细胞的增殖和集落形成能力. Transwell试验证明,新霉胺可阻滞A375细胞,乃至血管生成素诱导的A375细胞的迁移和侵润能力.此外,免疫荧光揭示新霉胺可阻断血管生成素的核转位,从而抑制血管生成素诱导的A375细胞增殖.上述结果提示,新霉胺可通过抑制血管生成素的核转位,从而抑制肿瘤细胞增殖、迁移和侵润.鉴于与新霉素比较,新霉胺毒性小,因此新霉胺可望作为黑色素瘤治疗的先导药物,颇具开发前景和潜力.     

Up-Regulation of Cdc37 Contributes to Schwann Cell Proliferation and Migration After Sciatic Nerve Crush

Cell division cycle protein 37 (Cdc37), a molecular chaperone takes part in a series of cellular processes including cell signal transduction, cell cycle progression, cell proliferation, cell motility, oncogenesis and malignant progression. It can not only recruit immature protein kinases to HSP90 but also work alone. Cdc37 was reported to be associated with neurogenesis, neurite outgrowth, axon guidance and myelination. However, the roles of Cdc37 on Schwann cells (SC) after peripheral nerve injury (PNI) remain unknown. In this study, we found that the expression of Cdc37 increased and reached the peak at 1 week after sciatic nerve crush (SNC), which was consistent with that of proliferation cell nuclear antigen. Immunofluorescence verified that Cdc37 co-localized with SC in vivo and in vitro. Intriguingly, Cdc37 protein level was potentiated in the model of TNF-α-induced SC proliferation. Moreover, we found that Cdc37 silencing impaired proliferation of SC in vitro. Moreover, Cdc37 suppression attenuated kinase signaling pathways of Raf–ERK and PI3K/AKT which are crucial cell signaling for SC proliferation. Finally, we found that Cdc37 silencing inhibited SC migration in vitro. In conclusion, we demonstrated that the way Cdc37 contributed to SC proliferation is likely via activating kinase signaling pathways of Raf–ERK and PI3K/AKT, and CDC37 was also involved in SC migration after SNC.     

泛素偶联酶2C促进肺癌细胞增殖和迁移与抑制细胞衰老

泛素偶联酶2C与多种肿瘤细胞的增殖密切相关,但其与肺癌发生和发展的关系尚不明确。 本研究以肺癌A549细胞为材料,通过RT-PCR、Western印迹、免疫荧光、SA-β-Gal细胞衰老染色、细胞划痕和Trans-well实验,阐明UBE2C与肺癌细胞的增殖、衰老和迁移能力的关系。结果显示,UBE2C在肺癌细胞中的表达明显高于正常细胞。利用基因修饰技术瞬时过表达或靶向沉默UBE2C后,在肺癌A549细胞中,UBE2C的mRNA和蛋白质水平显著增加3.5倍或减少0.5倍,显著促进或抑制细胞增殖,进而减少或增加细胞的凋亡率。过表达UBE2C后,显著抑制细胞衰老;但沉默UBE2C后,则增加细胞衰老。此外,过表达UBE2C后,下调转移相关基因E-钙黏着蛋白的mRNA和蛋白质表达水平,且上调波形蛋白基因的表达水平,进而促进肺癌细胞的迁移。但靶向敲除UBE2C后,上调E-钙黏着蛋白,同时下调波形蛋白表达水平,进而抑制肺癌细胞的迁移。本研究的开展将明确UBE2C在肺癌中的作用及其机制,为以UBE2C为靶点,提高病人生存期提供了理论基础。