المؤلفون: CHUNJING LIN¹, ZIYANG HUANG¹, RIYONG ZHOU¹, YING ZHOU¹, YANGPING SHENTU¹, KANG YU¹, YU ZHANG¹

المصدر: Experimental & Therapeutic Medicine. Feb2021, Vol. 21 Issue 2, p1-10. 10p.

مصطلحات موضوعية: *CELL migration, *CELL migration inhibition, *NITRIC-oxide synthases, *HELA cells, *CELL lines, *CELL proliferation

مستخلص: Notch3 is a member of the Notch family and its mutations are known to cause a hereditary human disorder called cerebral autosomal-dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL). However, the specific function and signaling cascade initiated by CADASIL mutants remain unknown. To gain further insight into mechanism of action of CADASIL mutants, the present study conducted several experiments on the effects of Notch3 mutants in multiple cell lines. The protein levels of Notch3, fibronectin, collagen, inducible nitric oxide synthase and DNA (cytosine-5)-methyltransferase 1 (DNMT1) were determined by western blotting. The mRNA levels of IL-1β and TNF-α were measured by reverse transcription semi-quantitative PCR and DNMT1 mRNA levels were determined by quantitative PCR. Trypan blue staining was used for proliferation analysis and wound healing assays were performed to determine cell migration capability. The present study reported that R90C and R169C Notch3 mutants, and wild-type Notch3 had different effects on several cell lines. In T/GHA-VSMC cells, following the transfection of the two mutants, collagen and fibronectin expression increased, whereas expression decreased in IMR-90 cells. In BV2 cells, the two mutants resulted in decreased nitric oxide and iNOS production. In HeLa cells, proliferation and migration increased significantly following the transfection of the two mutants, whereas in the MCF-7 and HCC1937 cell lines, cell proliferation and migration decreased. In addition, the two mutants suppressed the expression of DNMT1 in HeLa and IMR-90 cells. Overall, the present study provided novel insights that further explored the underlying mechanisms of CADASIL. [ABSTRACT FROM AUTHOR]

المؤلفون: SHUFANG HE¹, GUIYUAN YU¹, KE PENG¹, SISUN L¹ liusisun656@163.com

المصدر: Molecular Medicine Reports. Dec2020, Vol. 22 Issue 6, p5282-5292. 11p.

مصطلحات موضوعية: *TUMOR suppressor genes, *CELL migration inhibition, *HELA cells, *MESSENGER RNA, *CERVICAL cancer, *CELLS, *MICRORNA

مستخلص: MicroRNAs (miRs) can affect the progression of cervical cancer (CC). The present study investigated the function of miR‑145‑5p in CC and demonstrated its association with fascin (FSCN1). The expression levels of miR‑145‑5p in CC tissues and cell lines were analyzed using reverse transcription‑quantitative PCR, and its direct targets were explored using a luciferase reporter assay. The viability, migration and invasion of HeLa cells transfected with small interfering FSCN1 or with miR‑145‑5p mimics and inhibitors were analyzed using Cell Counting Kit‑8 and Transwell assays. The expression levels of FSCN1 mRNA and protein were investigated using reverse transcription PCR and western blotting. miR‑145‑5p was downregulated in CC tissues and cell lines. Moreover, overexpression of miR‑145‑5p inhibited the migration, invasion and viability of HeLa cells. miR‑145‑5p directly targeted FSCN1, which regulated the suppressive functions of miR‑145‑5p in CC cells. Overall, miR‑145‑5p is a tumor suppressor gene and a promising target for CC treatment. [ABSTRACT FROM AUTHOR]

المؤلفون: Miao, Juan¹ (AUTHOR), Zhu, Yiqing² (AUTHOR), Xu, Lei¹ (AUTHOR), Huang, Xiaohao¹ (AUTHOR) xiaohao512006@126.com, Zhou, Xue² (AUTHOR) maggiezhouxue840@126.com

المصدر: Molecular Medicine Reports. Nov2020, Vol. 22 Issue 5, p4442-4451. 10p.

مصطلحات موضوعية: *CELL migration inhibition, *CELL migration, *MICRORNA, *WESTERN immunoblotting, *PATHOLOGY

مستخلص: Normal placentation and successful maintenance of pregnancy depend on the successful migration and invasion of trophoblasts into maternal tissues. Previous studies reported that microRNAs (miRs) are expressed in trophoblasts, and can regulate their migration and invasion. The present study aimed to investigate miR-181b-5p function in HTR-8/SVneo trophoblasts and explore its underlying mechanism in the pathogenesis of multiple abnormal trophoblast invasion-related events. Reverse-transcription quantitative PCR and western blotting were used to test the expression of miR-181b-5p and sphingosine-1-phosphate receptor 1 (S1PR1) in samples of multiple abnormal trophoblast invasion-related events. Transwell invasion and wound healing assays were performed to determine cell invasion and migration abilities. A luciferase reporter assay was conducted to identify the downstream target of miR-181b-5p. Overexpression of miR-181b-5p suppressed HTR-8/SVneo cell migration and invasion, whereas inhibition of miR-181b-5p induced an opposite effect. The S1PR1 gene was further identified as a novel direct target of miR-181b-5p. Specifically, miR-181b-5p bound directly to the 3′-untranslated region of S1PR1 and suppressed its expression. Moreover, overexpression of S1PR1 reversed the inhibitory effect of miR-181b-5p. Taken together, ectopic expression of miR-181b-5p impaired the migration and invasion of trophoblasts by directly targeting S1PR1, thereby providing new insights into the pathogenesis of multiple abnormal trophoblast invasion-related events. [ABSTRACT FROM AUTHOR]

المؤلفون: Huang, Xiaohua¹ (AUTHOR), Li, Ying² (AUTHOR), He, Xin³ (AUTHOR), Chen, Yang¹ (AUTHOR), Wei, Wei¹ (AUTHOR), Yang, Xuesong³ (AUTHOR), Ma, Keli³ (AUTHOR) makelipaper@163.com

المصدر: Molecular Medicine Reports. Nov2020, Vol. 22 Issue 5, p3994-4002. 9p.

مصطلحات موضوعية: *CELL migration inhibition, *TYROSINE, *EPIDERMAL growth factor receptors, *GANGLIOSIDES, *COLON cancer, *CELL motility, *THIN layer chromatography

مستخلص: Previous studies have shown that (GM3), a ganglioside, suppresses hepatoma cell motility and migration by inhibiting phosphorylation of EGFR and the activity of the PI3K/AKT signaling pathway. Therefore, the aim of the present study was to investigate whether the combined treatment of CD82 with gangliosides can exert a synergistic inhibitory effect on cell motility and migration. Epidermal growth factor receptor (EGFR) signaling was studied for its role in the mechanism through which CD82 and gangliosides synergistically inhibit the motility and migration of SW620 human colon adenocarcinoma cells. GM3 and/or GM2 treatment, and/or overexpression of CD82 was performed in SW620 cells. High-performance thin layer chromatography, reverse transcription-quantitative PCR, western blotting and flow cytometry assays were used to confirm the content changes of GM2, GM3 and CD82. In addition, the phosphorylation of EGFR, MAPK and Akt were evaluated by western blot analysis. SW620 cell motility was investigated using wound healing analysis and chemotaxis migration assay. The combination of GM3 and GM2 with CD82 was found to markedly suppress EGF-stimulated SW620 cell motility compared with the individual factors or combination of GM2 or GM3 with CD82 by inhibiting the phosphorylation of EGFR. The results suggested that CD82 in combination with either GM2 or GM3 can exert a synergistic inhibitory effect on cell motility and migration; however, the synergistic mechanisms elicited by GM2 or GM3 with CD82 differ. [ABSTRACT FROM AUTHOR]

المؤلفون: Luo, Shunxiang¹ (AUTHOR), Shen, Ming¹ (AUTHOR), Chen, Hui¹ (AUTHOR), Li, Weiwei¹ (AUTHOR), Chen, Cong¹ (AUTHOR) qbdruq@163.com

المصدر: Molecular Medicine Reports. Nov2020, Vol. 22 Issue 5, p3822-3832. 11p.

مصطلحات موضوعية: *NON-small-cell lung carcinoma, *CELL migration inhibition, *ANTISENSE RNA, *WESTERN immunoblotting, *LUNG cancer

مستخلص: Non-small cell lung cancer (NSCLC) is a leading subtype of lung cancer, with high mortality rates. Recently, long non-coding RNAs (lncRNAs) have been associated with NSCLC. The present study aimed to examine the role of the TP73 antisense RNA 1 (TP73-AS1) lncRNA in NSCLC. TP73-AS1 and microRNA(miR)-34a-5p expression levels were measured using reverse transcription-quantitative PCR (RT-qPCR) and chromogenic in situ hybridization (CISH). Cell proliferation, apoptosis, migration and invasion was determined using Cell Counting Kit-8 (CCK-8), flow cytometry, Transwell and Matrigel assays, respectively. The median inhibitory concentration (IC50) value of cisplatin (cis-diamminedichloroplatinum; DDP) was assessed using a CCK-8 assay. The interaction between miR-34a-5p and TP73-AS1 or tripartite motif-containing 29 (TRIM29) was predicted using microRNA.org and Starbase, then verified using a dual-luciferase reporter assay. The expression of TRIM29 was quantified at the mRNA and protein level using RT-qPCR and western blot analysis, respectively. TP73-AS1 was significantly upregulated, while miR-34a-5p was downregulated in NSCLC tissues and cells. Functionally, TP73-AS1 knockdown inhibited proliferation, migration, invasion and DDP resistance, whilst inducing apoptosis in NSCLC cells. miR-34a-5p was identified as a target for TP73-AS1, and its inhibition reversed the effects of TP73-AS1 knockdown on NSCLC cells. In addition, TRIM29 was targeted by miR-34a-5p, and its overexpression reversed the effects of miR-34a-5p. Moreover, TP73-AS1 acted as a molecular sponge for miR-34a-5p, increasing the expression of TRIM29. In conclusion, TP73-AS1 contributed to proliferation, migration and DDP resistance but inhibited apoptosis of NSCLC cells by upregulating TRIM29 and sponging miR-34a-5p. [ABSTRACT FROM AUTHOR]

المؤلفون: Pan, Yuejiang¹ (AUTHOR), Jin, Ke¹ (AUTHOR), Xie, Xuan¹ (AUTHOR), Wang, Kexi¹ (AUTHOR), Zhang, Huizhong¹ (AUTHOR) zhhzhgd_2008@163.com

المصدر: Experimental & Therapeutic Medicine. Sep2020, Vol. 20 Issue 3, p2252-2261. 10p.

مصطلحات موضوعية: *NON-small-cell lung carcinoma, *CELL migration inhibition, *VASCULOGENIC mimicry, *CELL proliferation, *LUCIFERASES

مستخلص: MicroRNAs (miRNAs) are increasingly recognized as important regulators of non-small cell lung cancer (NSCLC) progression by directly regulating their target genes. The aim of the present study was to assess the biological role of miR-19a-3p in NSCLC. It was revealed that miR-19a-3p expression was significantly downregulated in human NSCLC tissues and cell lines compared with normal tissues and lung epithelial cells. In addition, a lower miR-19a-3p expression was significantly associated with Tumor Node Metastasis stage and lymph node metastasis. Furthermore, the upregulation of miR-19a-3p in NSCLC cell lines significantly inhibited cell proliferation, migration and invasion, as determined using an MTT, colony formation, wound healing and transwell Matrigel invasion assays, respectively. A luciferase reporter assay and western blotting determined that ubiquitin associated protein 2 like (UBAP2L) was a direct target of miR-19a-3p and could be inhibited through the upregulation of miR-19a-3p in NSCLC. In addition, UBAP2L silencing induced similar effects to those observed following miR-19a-3p overexpression. The overexpression of UBAP2L partially reversed the effects of miR-19a-3p on NSCLC cell lines. Collectively, these data indicated that miR-19a-3p may serve as a tumor suppressor partly through the regulation of UBAP2L expression in NSCLC and that the targeting of miR-19a-3p may be a novel method for NSCLC treatment. [ABSTRACT FROM AUTHOR]

المؤلفون: Xu, Yuan¹ (AUTHOR), Ma, Yuan² (AUTHOR), Liu, Xiao-Ling¹ (AUTHOR), Gao, Sheng-Li³ (AUTHOR)

المصدر: Molecular Medicine Reports. Jul2020, Vol. 22 Issue 1, p67-76. 10p.

مصطلحات موضوعية: *RENAL cell carcinoma, *CELL migration inhibition, *PROLIFERATING cell nuclear antigen, *CELL proliferation

مستخلص: Renal cell carcinoma has the highest incidence rate of cancer types in the urinary system. Moreover, microRNAs (miRNA) have been closely associated with numerous types of tumor. The present study aimed to investigate the effects of miRNA (miR)-133b on the proliferation, invasion and chemosensitivity of renal cell carcinoma cells, and to determine whether its mechanism was regulated by the ERK signaling pathway. Both renal cell carcinoma and adjacent healthy tissues from 60 patients, in addition to renal cell carcinoma lines, ACHN, Caki-1, A-498 and 786-O, and 293 cells, were used in this study. miR-133b expression was measured from renal cell carcinoma, adjacent healthy tissues and renal cell carcinoma cell lines by reverse transcription-quantitative PCR. Cells were transfected with miR-133b mimic to achieve miR-133b overexpression. The proliferative, migratory and invasive ability of the cells were evaluated using MTT, wound healing and Matrigel assays, respectively, and flow cytometry was used to detect the apoptotic rate. Following treatment with an ERK inhibitor, U0126, and activator, LM22B-10, western blotting was used to detect the expression of related proteins and the activity of the ERK signaling pathway. The overexpression of miR-133b significantly inhibited cell proliferation, migration and invasion, whilst inducing apoptosis and increasing the drug sensitivity of renal cell carcinoma cells to cisplatin, docetaxel and doxorubicin. The miR-133b mimic also increased the protein expression levels of Bax and decreased the expression levels of matrix metalloproteinase (MMP)-2, MMP-9, ATP-binding cassette subfamily G2, P-glycoprotein, Bcl-2 and proliferating cell nuclear antigen, as well as the phosphorylation of ERK (P<0.05). The administration of the U0216 inhibitor demonstrated similar effects to miR-133b overexpression, and there was no significant difference compared with the miR-133b mimic transfection (P>0.05). However, the overexpression of miR-133b combined with LM22B-10 treatment weakened the anticancer effects of miR-133b mimic transfection (P<0.05). In conclusion, miR-133b overexpression was observed to inhibit the proliferation, migration and invasion of renal cell carcinoma cells and improve chemotherapeutic sensitivity; it was suggested that the mechanism maybe related to the inhibition of ERK1/2 phosphorylation and thus decreased ERK signaling pathway activity. [ABSTRACT FROM AUTHOR]

المؤلفون: Chen, Yafei¹ (AUTHOR), Chen, Xin¹ (AUTHOR), Ding, Xiaojun¹ (AUTHOR), Wang, Yingwei¹ (AUTHOR) yingweiw_wyw@163.com

المصدر: Molecular Medicine Reports. Oct2019, Vol. 20 Issue 4, p3317-3325. 9p.

مصطلحات موضوعية: *CELL migration inhibition, *VASCULAR endothelial growth factors, *EPIDERMAL growth factor receptors, *WESTERN immunoblotting, *NEOPLASTIC cell transformation, *THERAPEUTIC embolization

مستخلص: Transcatheter arterial embolization (TAE) therapy has been used in the treatment of inoperable hepatocellular carcinoma (HCC). However, tumor recurrence and metastasis are common in patients after TAE, and these processes may be caused by circulating tumor cells (CTCs). Epithelial-mesenchymal transition (EMT) serves important roles in CTCs, and abnormal expression and activation of epidermal growth factor receptor (EGFR) is common in cancer cells. Afatinib is an EGFR-tyrosine kinase inhibitor (TKI). The present study aimed to investigate the effects of afatinib on EMT and tumorigenesis in HCC cells. Western blot analysis suggested that afatinib was able to effectively suppress overactivation of EGFR. Moreover, the expression levels of EMT- and metastasis-associated genes were found to be modulated by afatinib through EGFR inhibition. In addition, Cell Counting Kit-8 and Transwell assays suggested that the viability, migration and invasion of HCC cells were inhibited by afatinib through EGFR inhibition. Furthermore, the activity of the ERK signaling pathway and the expression levels of vascular endothelial growth factor (VEGF) and matrix metalloproteinase 9 (MMP9) were decreased following treatment with afatinib in vitro. Collectively, the present results suggested that the inhibitory effects of afatinib on EMT and tumorigenesis may be associated with the ERK-VEGF/MMP9 signaling pathway. The present study provides new insights into understanding the mechanism underlying HCC and may facilitate the development of novel therapeutic strategies to treat HCC recurrence. [ABSTRACT FROM AUTHOR]

المؤلفون: Xiong, Yan¹ (AUTHOR), Wang, Qibai² (AUTHOR) wangqibosci@163.com

المصدر: Molecular Medicine Reports. Oct2019, Vol. 20 Issue 4, p3055-3064. 10p.

مصطلحات موضوعية: *CELL migration inhibition, *WESTERN immunoblotting, *CANCER invasiveness, *TRANSFORMING growth factors, *GLIOBLASTOMA multiforme, *CANCER cell migration, *SMAD proteins

مستخلص: Stanniocalcin-1 (STC1) is involved in cancer progression; however, the function of STC1 in glioblastoma remains unknown. In the present study, the expression levels of STC1 protein in glioblastoma were detected using immunohistochemistry. The expression levels of STC1, SMAD2/3 and SMAD4 proteins, following silencing of STC1, were assessed via western blotting. EdU and Transwell assays were performed to determine the proliferation and migration ability of the cells. The mRNA expression levels of STC1, SMAD4 and microRNA (miR)-34a were determined using quantitative PCR. The expression levels of STC1 were increased in glioblastoma tissues. STC1 revealed a significant association with poor outcome in patients with glioblastoma (P<0.05). The proliferation and invasion abilities were repressed in LN229 cells infected with LV3-shSTC1-1 and LV3-shSTC1-2 compared with LV3-NC. By contrast, the proliferation and invasion abilities were increased in T98G cells infected with LV5-STC1 compared with LV5-NC (P<0.05). The expression levels of STC1, SMAD2/3 and SMAD4 were decreased in LN229 cells infected with LV3-shSTC1-1 and LV3-shSTC1-2 compared with LV3-NC. However, the expression levels of STC1, SMAD2/3 and SMAD4 were elevated in T98G cells infected with LV5-STC1 compared with LV5-NC. The expression levels of miR-34a were decreased following silencing of STC1 (P<0.05). The expression levels of SMAD4 were decreased when transfected with miR-34a mimics (P<0.05). The luciferase activity of the wild-type 3′untranslated region of SMAD4 was decreased following transfection with miR-34a mimics (P<0.05). Silencing of STC1 inhibited the growth of LN229 in vivo. In conclusion, STC1 expression levels were increased in the present study, and it was revealed that STC1 regulated glioblastoma malignancy. This phenotype was observed in the SMAD2/3 and SMAD4 pathways. [ABSTRACT FROM AUTHOR]

المؤلفون: Liang, Lingling^1,2 (AUTHOR), Wang, Xiaomei² (AUTHOR), Zheng, Yajuan¹ (AUTHOR) zhengyajuan124@126.com, Liu, Yang³ (AUTHOR) yangliu_manu@163.com

المصدر: Molecular Medicine Reports. Sep2019, Vol. 20 Issue 3, p2929-2935. 7p.

مصطلحات موضوعية: *CELL migration inhibition, *EXTRACELLULAR matrix, *APOPTOSIS, *TRANSFORMING growth factors-beta, *FILTERING surgery, *FIBROBLASTS, *APOPTOSIS inhibition, *WESTERN immunoblotting

مستخلص: Conjunctival fiber generation is implicated in a wide spectrum of ocular diseases. Conjunctival wound healing is characterized by inflammation followed by re-epithelialization, synthesis of new extracellular matrix (ECM), wound contraction and subconjunctival scar formation. The primary cause for the failure of glaucoma filtration surgery results from the excessive scarring of the filtering bleb. All-trans-retinoic acid (ATRA), a derivative of vitamin A, is a potent regulator of ECM synthesis, growth and differentiation. Following a previous study, which revealed that ATRA could inhibit transforming growth factor-β-induced human conjunctival fibroblast (HConF)-mediated collagen gel contraction, the present study aimed to investigate the effects of ATRA on HConF migration, apoptosis, proliferation and ECM synthesis. To achieve this, the present study used Transwell migration, wound healing and Cell Counting Kit-8 assays, flow cytometry and western blot analysis. In addition, the present study aimed to elucidate the mechanism of ATRA in mediating resistance to conjunctival scar formation. ATRA treatment resulted in an increased level of HConF apoptosis, reduced proliferation and migration, decreased collagen I and fibronectin expression, and decreased phosphorylation of PI3K and AKT. Thus, the present study showed a role for ATRA in inhibiting HConF migration, proliferation and ECM synthesis, and in promoting HConF apoptosis through the inhibition of the PI3K/AKT signaling pathway. [ABSTRACT FROM AUTHOR]