SLC38A1 Promotes Proliferation and Migration of Human Colorectal Cancer Cells

Current studies have demonstrated that SLC38A1 proteins play a causal role in neoplastic cell transformation. The twofold aim of this study was to provide insight into whether a variance in the expression of SLC38A1 exists between human colorectal cancer and healthy human tissues and to determine how silencing or overexpressing the SLC38A1 gene could affect the proliferation, viability and migration of colorectal cancer cells. Immunohistochemical staining was used to analyze the expression of SLC38A1 in colorectal cancer tissues and adjacent normal mucosa in 77 patients who underwent surgical resection. The expression of SLC38A1 in colorectal cancer tissues and cell lines was detected using RT-PCR and Western blotting. Two colorectal cancer cell lines SW480 and HCT116 were used to examine whether silencing SLC38A1 with si RNA and overexpressing SLC38A1 with sh RNA could affect cell viability and migration. As a result, the SLC38A1 protein was very low or undetectable in the normal colon mucosa. In contrast, strong staining of SLC38A1 protein was found in the cytoplasm in 79.2% colorectal cancer samples. More pronounced SLC38A1 expression in colorectal cancer tissues was significantly associated with tumor node metastasis（TNM） stage. Inhibition of SLC38A1 reduced tumour growth and suppressed proliferation and migration of SW480 cells. In contrast, overexpression of SLC38A1 had the opposite effects on HCT116 cells. SLC38A1 is overexpressed in colorectal cancer, which suggests that it is associated with tumour progression. These results encourage the exploration of SLC38A1 as a target for intervention in colorectal cancer.

RNAi-mediated Human Nestin Silence Inhibits Proliferation and Migration of Malignant Melanoma Cells by G1/S Arrest via Akt-GSK3β-Rb Pathway

Human Nestin（hNestin） has been found to express in melanoma, and its expression is positively correlated with the advanced stage of melanoma. However, the precise role of hNestin in the development of melanoma has not been fully understood. The present study aimed to explore the role of hNestin in the proliferation and invasion of melanoma cells. The lentivirus vector carrying a short hairpin RNAs（shRNAs） targeting hNestin（hNestin-sh RNA-LV） was stably infected into human melanoma cells UACC903, which expressed high levels of hNestin. The effects of hNestin knockdown on the proliferation, apoptosis, migration of melanoma cells and the related signaling pathways were investigated by immunofluorence, Western blotting and reverse transcription polymerase chain reaction（RT-PCR）, respectively. The results showed that hNestin was expressed in most melanoma specimens and the melanoma cells studied. Knockdown of hNestin expression significantly inhibited the proliferation of melanoma cells, blocked the formation of cell colony, arrested cell cycle at G1/S stage and suppressed the activation of Akt and GSK3β. hNestin-silent cells also showed a sheet-like appearance with tight cell-cell adhesion, decreased membrane expression of N-cadherin and β-catenin, and attenuated migration. Furthermore, hNestin silence resulted in the inhibition of tumor growth in vivo. Our study indicates that hNestin knockdown suppresses the proliferation of melanoma cells, which might be through affecting Akt-GSK3β-Rb pathway-mediated G1/S arrest, and hNestin silence inhibits the migration by selectively modulating the expression of cell adhesion molecules in the process of epithelial-mesenchymal transition.

Effect of Endostar combined with Paclitaxel on the proliferation and migration of metalloproteinases and tumor cells in NSCLC patients

Objective: To explore the effect of endostar combined with taxol on tumor markers, vascular endothelial growth factor, neuron-specific enolase, metalloproteinase and tumor cell proliferation and migration in NSCLC patients. Methods Patients with advanced NSCLC were studied. The patients in the control group received chemotherapy with paclitaxel combined with cisplatin. Patients in the combination group received intravenous infusion of endostar on the basis of treatment of patients in the control group. 21 days was a cycle, and all patients were treated for 2 cycles. Fasting venous blood 3mL of all patients before and after treatment was collected, and CEA and saccharide antigen (CA50) were detected by radioimmunoassay. Vascular endothelial growth factor (VEGF), neuron-specific enolase (NSE), serum matrix metalloproteinase (MMP-2, MMP-9), high-mobility family protein at-hook 2 (HMGA 2) and high-mobility family protein B 1 (HMGB 1) were detected by ELISA. Results There were no significant differences in serum CA50, CEA, VEGF, NSE, mmp-2, mmp-9, HMGB 1, and HMGA 2 between the two groups before treatment (P>0.05). After two courses of chemotherapy, CA50, CEA, VEGF, NSE, MMP-2, MMP-9, HMGB 1, and HMGA 2 in the combination group and control group were significantly lower than before treatment (P<0.05), and the combination group was significantly lower than the control group (P<0.05). Conclusion Endostar combined with paclitaxel can enhance the chemotherapy effect of NSCLC patients, reduce the level of serum tumor markers, neuronal specific enolase and vascular endothelial growth factor, and inhibit the proliferation and migration of tumor cells.

A human-specific insertion promotes cell proliferation and migration by enhancing TBC1D8B expression

Human-specific insertions play important roles in human phenotypes and diseases.Here we reported a 446-bp insertion(Insert-446)in intron 11 of the TBC1D8B gene,located on chromosome X,and traced its origin to a portion of intron 6 of the EBF1 gene on chromosome 5.Interestingly,Insert-446 was present in the human Neanderthal and Denisovans genomes,and was fixed in humans after human-chimpanzee divergence.We have demonstrated that Insert-446 acts as an enhancer through binding transcript factors that promotes a higher expression of human TBC1D8B gene as compared with orthologs in macaques.In addition,over-expression TBC1D8B promoted cell proliferation and migration through“a dual finger”catalytic mechanism(Arg538 and Gln573)in the TBC domain in vitro and knockdown of TBC1D8B attenuated tumorigenesis in vivo.Knockout of Insert-446 prevented cell proliferation and migration in cancer and normal cells.Our results reveal that the human-specific Insert-446 promotes cell proliferation and migration by upregulating the expression of TBC1D8B gene.These findings provide a significant insight into the effects of human-specific insertions on evolution.

Monocyte chemoattractant protein 1 and fractalkine play opposite roles in angiogenesis via recruitment of different macrophage subtypes

AIM： To explore the interaction between macrophages and chemokines [monocyte chemoattractant protein 1（MCP-1/CCL2） and fractalkine/CX3CL1] and the effects of their interaction on neovascularization.METHODS： Human peripheral blood mononuclear cells, donated by healthy volunteers, were separated and cultured in RPMI-1640 medium containing 10% fetal bovine serum, then induced into macrophages by stimulation with 30 μg/L granulocyte macrophage-colony stimulating factor（GM-CSF）. The expression of CCR2 and/or CX3CR1 in the macrophages was examined using flow cytometry. Macrophages were then stimulated with recombinant human CCL2（rh-CCL2） or recombinant human CX3CL1（rh-CX3CL1）. The expression of angiogenesis-related genes, including VEGF-A, THBS-1 and ADAMTS-1 were examined using real-time quantitative polymerase chain reaction（PCR）. Supernatants from stimulated macrophages were used in an assay of human retinal endothelial cell（HREC） proliferation. Finally, stimulated macrophages were cocultured with HREC in a migration assay.RESULTS： The expression rate of CCR2 in macrophages stimulated by GM-CSF was 42%±1.9%. The expression rate of CX3CR1 was 71%±3.3%. Compared with vehicle-treated groups, gene expression of VEGF-A in the macrophages was greater in 150 mg/L CCL2-treated groups（P〈0.05）, while expression of THBS-1 and ADAMTS-1 was significantly lower（P〈0.05）. By contrast, compared with vehicle-treated groups, expression of VEGF-A in 150 mg/L CX3CL1-treated groups was significantly lower（P〈0.05）, while expression of THBS-1 and ADAMTS-1 was greater（P〈0.05）. Supernatants from CCL2 treated macrophages promoted proliferation of HREC（P〈0.05）, while supernatants from CX3CL1-treated macrophages inhibited the proliferation of HREC（P〈0.05）. HREC migration increased when co-cultured with CCL2-treated macrophages, but decreased with CX3CL1-treated macrophages（P〈0.05）.CONCLUSION： CCL2 and CX3CL1 exert different effects in regulation of macrophage in expression of angiogenesisrelated factors, including VEGF-A, THBS-1 and ADAMTS-1. Our findings suggest that CCL2 and CX3CL1 may be candidate proteins for further exploration of novel targets for treatment of ocular neovascularization.

Knockdown of GRHL3 Inhibits Activities and Induces Cell Cycle Arrest and Apoptosis of Human Colorectal Cancer Cells

The Grainyhead-like 3（GRHL3） is involved in epidermal barrier formation, neural tube closure and wound repair. Previous studies have suggested that GRHL3 has been linked to many different types of cancers. However, to date, its effects on human colorectal cancer（CRC） has not been clarified yet. Our microarray analysis has indicated predominant GRHL3 expression in CRC. The purpose of this study was to investigate the expression and significance of GRHL3 in CRC tumorigenesis using CRC tissues and paired paracancerous tissues, as well as using distinct CRC cell lines（HT29 and DLD1）. We observed increased GRHL3 expression at both m RNA and protein levels in CRC tissues and CRC cell lines using quantitative real-time polymerase chain reaction（q RT-PCR） and Western blotting. Moreover, silencing GRHL3 with si RNA could suppress CRC cell proliferation, viability and migration in vitro. We also found that knockdown of GRHL3 could promote cell cycle arrest at G0/G1 phase in HT29 cells and DLD1 cells, and induce cell apoptosis in HT29 cells. Together, our study revealed the down-regulation of GRHL3 in vitro could inhibit CRC cell activity and trigger cell cycle arrest at G0/G1 phase and apoptosis.

MiR-29b suppresses the proliferation and migration of osteosarcoma cells by targeting CDK6

Osteosarcoma is the most common primary sarcoma of bone, and it is a leading cause of cancer death among adolescents and young adults. However, the molecular mechanism underlying osteosarcoma carcinogenesis remains poorly understood. Recently, cyclin-dependent kinase 6 （CDK6） was identified as an important onco- gene. We found that CDK6 protein level, rather than CDK6 mRNA level, is much higher in osteosarcoma tissues than in normal adjacent tissues, which indicates a post-transcriptional mechanism involved in CDK6 regulation in osteosarcoma. MiRNAs are small non- coding RNAs that repress gene expression at the post- transcriptional level and have widely been shown to play important roles in many human cancers. In this study, we investigated the role of miR-29b as a novel regulator of CDK6 using bioinformatics methods. We demon- strated that CDK6 can be downregulated by miR-29b via binding to the 3＇-UTR region in osteosarcoma cells. Furthermore, we identified an inverse correlation between miR-29b and CDK6 protein levels in osteosar- coma tissues. Finally, we examined the function of miR- 29b-driven repression of CDK6 expression in osteosarcoma cells. The results revealed that miR-29b acts as a tumor suppressor of osteosarcoma by targeting CDK6 in the proliferation and migration processes. Taken together, our results highlight an important role for miR-29b in the regulation of CDK6 in osteosarcoma and may open new avenues for future osteosarcoma therapies.

Role of oxysterol-binding protein-related proteins in malignant human tumours

The oxysterol-binding protein-related protein(ORP)family is a group of proteins that mediate oxysterol metabolism and bioactivity in cells.ORPs constitute a large family of lipid transfer proteins.Much of the current evidence indicates that certain members of the family of oxysterol-binding proteins(OSBPs)can lead to cancer.Many studies have revealed the putative roles of OSBPs in various cancer types.However,the exact effects and mechanisms of action of members of the OSBP/ORP family in cancer initiation and progression are currently unclear.This review focuses on ORP family members that can accelerate human tumour cell proliferation,migration,and invasion.The mechanisms and functions of various ORPs are introduced in detail.We also attempt to identify the roles of these proteins in malignant tumours with the ultimate aim of determining the exact role of the OSBP/ORP family in human tumour cells.

Effects of human microRNA-181a-5p on proliferation and migration of gastric cancer cells

Objective To preliminarily explore the effects of human microRNA-181a on migration of gastric cancer cells and its mechanism.Methods The expression of miRNA-181a-5p in gastric cancer cell line GC9811 and peritoneal high metastasis gastric cancer cell line GC9811-P were tested by quantitative real-time polymerase chain reaction(qRT-PCR).GC9811 cell line was

Photoelectric-driven conductive composite ionogel patch for effective wound healing

Developing the high biosafety,effective and wearable devices for fast wound healing is highly desired but remains a challenge.Here,we propose a“win–win co-operation”strategy to potentiate effective skin wound healing at the wound site by constructing robust and ecofriendly composite patch under opto-electric stimulation.The wearable patch is composed of ionic gel doped with Ti3C2Tx(MXene),which possesses good photothermal response to kill the bacteria via effective inhibition of the expression of inflammatory factors,preventing wound infection.Importantly,the composite ionogel patch is capable of providing green and on-demand electrical stimulation for wound site,guiding cell migration and proliferation by improved bioenergy and expression up-regulation of growth factor.In mice wound models,the treatment group healed~31%more rapidly.Mechanistically,the wearable devices could enable visual and real-time supervising treatment effect due to their good transmittance.The proposed strategy would be promising for future clinical treatment of wound healing.

Chungtaejeon, a Korean fermented tea, prevents the risk of atherosclerosis in rats fed a high-fat atherogenic diet

OBJECTIVE： Hypercholesterolemia is one of the well-established risk factors for cardiovascular mortality and morbidity in coronary heart disease. The aim of this study was to investigate the anti-atherogenic effect of Chungtaejeon（CTJ, a Korean fermented tea） aqueous extract on proliferation and migration of human aortic smooth muscle cells（HASMCs） in vivo and in vitro.METHODS： The authors used high-fat atherogenic diet（HFAD） to induce hyperlipidemia in Wistar rats in in vivo animal experiments and used HASMCs for in vitro cell experiments. For the in vitro cell experiment, the proliferation of HASMCs was evaluated using the MTT assay. Similarly, the expression of matrix metalloproteinases（MMPs） in HASMCs was measured using gelatin zymography. Antimigratory activity of CTJ was revealed using the wound-healing model and Boyden＇s chamber assay. In the in vivo experiment, CTJ was administered in three different doses for 20 d from the initiation of the HFAD. After 20 d, the serum lipid profile and total lipid contents in liver were measured.RESULTS： Treatment with CTJ for 24 h dose-dependently inhibited the proliferation and migration of HASMCs and expression of MMP-2 in HASMCs. The oral administration of CTJ at concentrations of 200 and 400 mg/kg decreased the levels of low-density lipoprotein cholesterol, total serum cholesterol and hepatic cholesterol of HFAD-fed rats.CONCLUSION： CTJ possessed strong antiproliferative, antimigratory, as well as lipid-lowering activities. Thus, CTJ can be considered as a therapeutic option in the treatment of high-fat diet-induced atherosclerosis.

Effects of hepatocyte growth factor-mediated activation of Dll4-Notch-Hey2 signaling pathway

Background Hepatocyte growth factor （HGF） treats ischemic disease by promoting arteriogenesis, however, its mechanism of action is not known. The notch signaling pathway plays an important role in neovascularization. The relationship between the proliferation and migration ability of artery endothelial cells and the Dll4-Notch-Hey2 signaling pathway in the process of arteriogenesis was investigated as a mechanism of action of HGE Methods Based on the prophase study cells and supernatant were harvested at the indicated time after human femoral artery endothelial cells （HFAECs） were infected with adenovirus-HGF （Ad-HGF） at 200 pfu/cell. Cells were analyzed for HGF expression and Notch1, DII4 and Hey2 expression by ELISA and reverse transcription-PCR （RT-PCR）. The changes in the proliferation and migration ability of HFAECs were observed by M-I-I- and Transwell migration experiments Ad-GFP-infected HFAECs were used as control. Results Compared with the control group the Ad-HGF group＇s HGF expression was not increased with time, and the induction by HGF of Notch1, DII4 and Hey2 gene transcription was not enhanced with an increase of HGF. The proliferation ability of Ad-HGF-transduced HFAECs was enhanced and their migration ability was also enhanced in the presence of HGF. Conclusions Through activating the DII4-Notch-Hey2 signaling pathway, HGF indirectly promotes the proliferation and migration ability of cells, so that offspring artery branches are formed.