Ubiquitin ligase TRIM46 promotes the proliferation and invasion of colorectal cancer cells北大核心CSCD

Objective: To explore the effect of tripartite motif 46 (TRIM46) on the proliferation and invasion of colorectal cancer cells and study its possible mechanism. Methods: After TRIM46 siRNA was transfected into human colorectal cancer HCT116 cells, the expressions of TRIM46 mRNA and protein were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. CCK-8 assay and Transwell chamber assay were implemented to observe the effect of TRIM46 down-regulation on the proliferation and invasion of HCT116 cells, and the expression of Src homology 2 (SH2)-containing tyrosine phosphatase 1 (SHP1) was detected by Western blotting. Results: After TRIM46 siRNA was transfected into human colorectal cancer HCT116 cells, the expression levels of TRIM46 mRNA and protein were inhibited (both P < 0.05). Knockdown of TRIM 46 significantly suppressed the cell proliferation and invasion of HCT116 cells (both P < 0.05). However, the expression level of SHP1 was increased (P < 0.05). Conclusion: Ubiquitin ligase TRIM46 may improve the proliferation and invasion of human colorectal cancer cells through suppressing SHP1. Copyright © 2017 by TUMOR All rights reserved.     

MACC1 promotes carcinogenesis of colorectal cancer via β-catenin signaling pathway

Here we confirmed that metastasis-associated in colon cancer 1 (MACC1) and β-catenin expression were higher in colorectal cancer (CRC) cells and tissues than those in normal colonic epithelial cell line and adjacent non-tumour colorectal mucosa (ANM) tissues, respectively. MACC1 expression was significantly related to histological differentiation (p<0.001), UICC stage (p=0.029), T classification (p=0.017), and N classification (p=0.023). Cox regression analysis demonstrated that high MACC1/abnormal β-catenin expression was the strongest independent prognostic indicator for reduced overall survival in CRC patients. Significant positive correlation between MACC1 expression and abnormal β-catenin expression was found in CRC tissues. MACC1 knockdown dramatically inhibited cellular proliferation, migration, invasion, colony formation, and tumorigenesis, both in vitro and in vivo, but induced apoptosis in CRC cells. Further MACC1 over-expression increased Met, β-catenin, and its downstream genes including c-Myc, cyclin D1, and MMP9 expression, and its upstream gene phos-GSK3β (Ser9) expression. In addition, MACC1 increased vimentin and suppressed E-cadherin in HCT116 cells. Silencing of MACC1 reversed all these changes. Our results firstly suggest that MACC1 plays an important role in carcinogenesis and progression of CRC through β-catenin signaling pathway and mesenchymal-epithelial transition.     

ARMC8α promotes proliferation and invasion of non-small cell lung cancer cells by activating the canonical Wnt signaling pathway

ARMC8 proteins are novel armadillo repeat containing proteins, which are well conserved in eukaryotes and are involved in a variety of processes such as cell migration, proliferation, tissue maintenance, signal transduction, and tumorigenesis. Armadillo repeat proteins include well-known proteins such as β-catenin and p120ctn. Our current knowledge of ARMC8, especially its role in cancer, is limited. In this study, we quantified ARMC8 expression in 112 non-small cell lung cancer (NSCLC) tissues and adjacent non-cancerous tissues, and seven lung cancer cell lines using immunohistochemistry staining and Western blotting. ARMC8 level was significantly higher in NSCLC tissues than in the adjacent normal tissues (67.9 % versus 5.4 %, p?p?=?0.022), lymph node metastasis (p?=?0.001), and poor prognosis (p?NSCLC patients. Cox regression analysis demonstrated that ARMC8 was an independent prognostic factor for NSCLC. Consistent with this, ARMC8α downregulation by siRNA knockdown inhibited growth, colony formation, and invasion in A549 lung cancer cells, while ARMC8α overexpression promoted growth, colony formation, and invasion in H1299 lung cancer cells. In addition, ARMC8α knockdown downregulated canonical Wnt-signaling pathway activity and cyclin D1 and matrix metalloproteinase (MMP)-7 expression. Consistent with this, ARMC8α overexpression upregulated canonical Wnt-signaling pathway activity and cyclin D1 and MMP-7 expression. These results indicate that ARMC8α upregulates cyclin D1 and MMP7 expression by activating the canonical Wnt-signaling pathway and thereby promoting lung cancer cell proliferation and invasion. Therefore, ARMC8 might serve as a novel therapeutic target in NSCLC.     

SDF-1/CXCR4 promotes epithelial–mesenchymal transition and progression of colorectal cancer by activation of the Wnt/β-catenin signaling pathway

Stromal cell-derived factor 1 (SDF-1) and its receptor, CXCR4, play an important role in angiogenesis and are associated with tumor progression. This study aimed to investigate the role of SDF-1/CXCR4-mediated epithelial–mesenchymal transition (EMT) and the progression of colorectal cancer (CRC) as well as the underlying mechanisms. The data showed that expression of CXCR4 and β-catenin mRNA and protein was significantly higher in CRC tissues than in distant normal tissues. CXCR4 expression was associated with β-catenin expression in CRC tissues, whereas high CXCR4 expression was strongly associated with low E-cadherin, high N-cadherin, and high vimentin expression, suggesting a cross talk between the SDF-1/CXCR4 axis and Wnt/β-catenin signaling pathway in CRC. In vitro, SDF-1 induced CXCR4-positive colorectal cancer cell invasion and EMT by activation of the Wnt/β-catenin signaling pathway. In contrast, SDF-1/CXCR4 axis activation-induced colorectal cancer invasion and EMT was effectively inhibited by the Wnt signaling pathway inhibitor Dickkopf-1. In conclusion, CXCR4-promoted CRC progression and EMT were regulated by the Wnt/β-catenin signaling pathway. Thus, targeting of the SDF-1/CXCR4 axis could have clinical applications in suppressing CRC progression.     

Nicotine upregulates microRNA-21 and promotes TGF-β-dependent epithelial-mesenchymal transition of esophageal cancer cells

A consistent positive association between cigarette smoking and the human esophageal cancer has been confirmed all over the world. However, details in the association need to be more focused on and be identified. Recently, aberrantly expressed microRNAs (miRNAs) have been shown to be promising biomarkers for understanding the tumorigenesis of a wide array of human cancers, including the esophageal cancer, and the deregulation on the epithelial to mesenchymal transition (EMT) by miRNAs is involved in the tumorigenesis. In present study, we were going to identify the role of nicotine-induced miR-21 in the EMT of esophageal cells. We found that there was an overexpression of miR-21 in esophageal specimens, having an association with cigarette smoking, and the upregulation of miR-21 was also induced by nicotine in esophageal carcinoma cell line, EC9706. Moreover, the upregulated miR-21 by nicotine promoted EMT transforming growth factor beta (TGF-β) dependently. Thus, the present study reveals a novel oncogenic role of nicotine in human esophageal cancer.     

PUS7 promotes the proliferation of colorectal cancer cells by directly stabilizing SIRT1 to activate the Wnt/β-catenin pathway

Pseudouridine synthase 7 (PUS7) may play key roles in cancer development. However, few studies have been conducted in this area. In the present study, we explored the function and potential mechanisms of PUS7 in colorectal cancer (CRC) progression. We found that PUS7 had higher expression in CRC tissues and cell lines. Clinically, high expression of PUS7 was associated with an unfavorable prognosis for CRC patients. Functionally, knockdown of PUS7 suppressed the proliferation of CRC cells in vitro and inhibited tumorigenicity in vivo. Mechanistically, RNA sequencing and coimmunoprecipitation (Co-IP) indicated that PUS7 exhibited oncogenic functions through the interaction of Sirtuin 1 (SIRT1) and activated the Wnt/β-catenin signaling pathway. Thus, our findings suggest that PUS7 promotes the proliferation of CRC cells by directly stabilizing SIRT1 to activate the Wnt/β-catenin pathway.     

LGR5 promotes the proliferation and tumor formation of cervical cancer cells through the Wnt/β-catenin signaling pathway

Leucine-rich repeat-containing G protein-coupled receptor 5 (LGR5), a seven transmembrane receptor known as a potential stem cell marker for intestinal crypts and hair follicles, has recently been found to be overexpressed in some types of human cancers. However, the role of LGR5 in cervical cancer remains unclear. In this study, the expression of LGR5 gradually increases from normal cervix to cervical cancer in situ and to cervical cancers as revealed by immunohistochemistry and western blot analyses. Through knocking down or overexpressing LGR5 in SiHa and HeLa cells, the expression level of LGR5 was found to be positively related to cell proliferation in vitro and to tumor formation in vivo. Further investigation indicated that LGR5 protein could significantly promote the acceleration of cell cycle. Moreover, the TOP-Flash reporter assay and western blot for β-catenin, cyclinD1, and c-myc proteins, target genes of the Wnt/β-catenin pathway, indicated that LGR5 significantly activated Wnt/β-catenin signaling. Additionally, the blockage of Wnt/β-catenin pathway resulted in a significant inhibition of cell proliferation induced by LGR5. Taken together, these results demonstrate that LGR5 can promote proliferation and tumor formation in cervical cancer cells by activating the Wnt/β-catenin pathway.     

Overexpression of CSRP1 gene promotes proliferation,migration and invasion of human lung adenocarcinoma H1299 cells北大核心CSCD

Objective: To investigate the effects of cysteine and glycine rich protein 1 (CSRP1) gene overexpression on proliferation, cell cycle, migration and invasion of human lung adenocarcinoma H1299 cells, and to explore the posssible mechanism. Methods: The recombinant lentivirus plasmids pCDH-CSRP1 carrying CSRP1 gene and pCDH-GFP (as the control) were constructed and infected into human lung adenocarcinoma H1299 cells, respectively. The expression levels of CSRP1 mRNA and protein in H1299 cells were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. The proliferation of H1299 cells with CSRP1 gene overexpression was tested by CCK-8 assay and colony formation assay. The cell cycle distribution of H1299 cells was detected by FCM. The longitudinal and lateral migration abilities of H1299 cells were detected by Transwell chamber assay and scratch wound healing assay. The invasion ability of H1299 cells was detected by Transwell chamber assay. The expression levels of focal adhesion kinase (FAK) and phospho-FAK (p-FAK) proteins in H1299 cells were detected by Western blotting. Results: The recombinant lentivirus plasmid pCDH-CSRP1 was successfully constructed, and the expression of CSRP1 in H1299 cells transfected with pCDH-CSRP1 was significantly higher than that in H1299 cells transfected with pCDH-GFP (P < 0.01). The overexpression of CSRP1 gene promoted the proliferation of H1299 cells (P < 0.05), and contributed to the increased proportion of G1 phase (P < 0.01) and the decreased proportion of S phase (P < 0.01) in H1299 cells. Furthermore, the overexpression of CSRP1 gene facilitated the migration and invasion abilities of H1299 cells (both P < 0.01). Compared with the control group, the expression level of p-FAK increased significantly (P < 0.01), while the expression level of FAK remained unchanged (P > 0.05) in H1299 cells with CSRP1 gene overexpression. Conclusion: CSRP1 gene overexpression can promote the proliferation, migration and invasion of human lung adenocarcinoma H1299 cells, in which the FAK signaling pathway may be involved. Copyright © 2018 by TUMOR. All rights reserved.     

CELLFOOD? induces apoptosis in human mesothelioma and colorectal cancer cells by modulating p53, c-myc and pAkt signaling pathways

Background

CELLFOOD™ (CF) is a nutraceutical non-addictive, non-invasive, and completely non-toxic unique proprietary colloidal-ionic formula. Little is known about its effect on cancer cells in solid tumors. The aim of this study was to evaluate the effect that CF has on different cancer cell lines and the mechanism by which the nutraceutical works.

Methods

The effect of CF on HFF (normal fibroblasts), Met5A (mesothelium), MSTO-211H, NCI-2452, Ist-Mes1, MPP89, Ist-Mes2 (mesothelioma), M14 (melanoma), H1650, H1975 (lung cancer), SKRB3 (breast cancer), and HCT-116 (colorectal cancer) cell growth was tested by cell proliferation and clonogenic assay. Among all of them, MSTO-211 and HCT-116 were analyzed for cell cycle by flow cytometry and western blot.

Results

All human cancer lines were suppressed on cell growth upon 1:200 CF treatment for 24 and 48 hours. Death was not observed in HFF and Met5A cell lines. Cell cycle analysis showed an increased sub-G1 with reduction of G1 in MSTO-211 and a cell cycle arrest of in G1 in HCT116. Activation of caspase-3 and cleavage of PARP confirmed an apoptotic death for both cell lines. Increased expression levels of p53, p21, and p27, downregulation of c-myc and Bcl-2, and inhibition of Akt activation were also found in CF-treated MSTO-211 and HCT-116 cells.

Conclusions

These findings ascertained an interaction between p53, c-myc, p21, p27, Bcl-2, PI3K/Akt pathway, and CF-induced apoptosis in MSTO-211H and HCT-116 cells, suggesting that CF acts as an important regulator of cell growth in human cancer cell lines. CF could be a useful nutraceutical intervention for prevention in colon cancer and mesothelioma.     

5-hydroxytryptamine receptor (5-HT1DR) promotes colorectal cancer metastasis by regulating Axin1/β-catenin/MMP-7 signaling pathway

Overexpression of 5-hydroxytryptamine (5-HT) in human cancer contributes to tumor metastasis, but the role of 5-HT receptor family in cancer has not been thoroughly explored. Here, we report overexpression of 5-HT_1D receptor (5-HT_1DR) was associated with Wnt signaling pathway and advanced tumor stage. The underlying mechanism of 5-HT_1DR-promoted tumor invasion was through its activation on the Axin1/β-catenin/MMP-7 pathway. In an orthotopic colorectal cancer mouse model, we demonstrated that a 5-HT_1DR antagonist ({"type":"entrez-nucleotide","attrs":{"text":"GR127935","term_id":"238377770","term_text":"GR127935"}}GR127935) effectively inhibited tumor metastasis through targeting Axin1. Furthermore, in intestinal epithelium cells, we observed that 5-HT_1DR played an important role in cell invasion via Axin1/β-catenin/MMP-7 pathway. Together, our findings reveal an essential role of the physiologic level of 5-HT_1DR in pulmonary metastasis of colorectal cancer.     

Hypoxia promotes migration and invasion of pancreatic cancer cells via inducing calpain 2 expression北大核心CSCD

Objective: To investigate the expressions of Calpain 1 and Calpain 2 proteins, as well as the effects of regulating Calpain 2 expression on the migration and invasion of pancreatic cancer PANC-1 cells in hypoxia microenvironment. Methods: The expressions of Calpain 1 and Calpain 2 mRNAs in pancreatic cancer tissues and their survival prognosis value were analyzed using GEPIA (Gene Expression Profiling Interactive Analysis) database online tool. The pancreatic cancer PANC-1 cells were cultured in normoxia (21% O2) and hypoxic (1% O2) environment, then the expressions of Calpain 1 and Calpain 2 mRNAs and proteins were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. The siRNA targeting CAPN2gene(encoding Calpain 2) was transfected into pancreatic cancer PANC-1 cells, then the silencing effect of CAPN2 gene was detected by real-time fluorescent quantitative PCR and Western blotting. The PANC-1 cells with Calpain 2 low expression were cultured under the normoxia and hypoxic conditions, then the migration and invasion abilities of these cells were detected by wound healing test and Transwell chamber assay, the expressions of E-cadherin and Vimentin were detected by Western blotting, and the cell morphology was observed by inverted microscopy. Results: The expressions of Calpain 1 and Calpain 2 mRNAs in human pancreatic cancer tissues were higher than those in normal pancreatic tissues (both P < 0.01), and the high expression of Calpain 2 was associated with the poor prognosis of pancreatic cancer patients (P = 0.013). Compared with the normoxia, the expression levels of Calpain 2 mRNA and protein were increased in hypoxia microenvironment (both P< 0.01), but the expressions of Calpain 1 mRNA and protein were not changed. The Calpain 2 low-expressed PANC-1 cells were established successfully. Under normoxia condition, the down-regulation of Calpain 2 expression inhibited the migration and invasion of PANC-1 cells (both P < 0.05); hypoxia promoted the migration and invasion of PANC-1 cells (both P < 0.05); but the down-regulation of Calpain 2 expression could reverse the promotion effects of hypoxia on the migration and invasion of PANC-1 cells (both P < 0.05). Furthermore, the expression of E-cadherin increased, the expression of Vimentin decreased in PANC-1 cells after the down-regualtion of Calpain 2 expression under normoxia condition (both P < 0.05), while the size of irregular cells increased, and the part of cells had apoptotic change; but the expression of E-cadherin decreased, the expression of Vimentin increased in PANC-1 cells cultured under hypoxia condition (both P < 0.05), while the most of cells had mesenchymal-like changes, showing long shuttle type; but the down-regulation of Calpain 2 expression could reverse the effect of hypoxia on the epithelial-mesenchymal transition of PANC-1 cells (both P< 0.05). Conclusion: Stimulated by hypoxia microenvironment, the expression of Calpain 2 was increased, so as to promote the migration and invasion of PANC-1 cells via regulating E-cadherin and Vimentin expressions. © 2019 by TUMOR. All rights reserved.     

SREBP2m promotes proliferation and migration of hepatocarcinoma cells and inhibits apoptosis through cholesterol metabolism damage北大核心CSCD

Objective: To investigate the effects of sterol regulatory element binding protein 2 (SREBP2) on the metabolism of cholesterol as well as the proliferation, apoptosis and migration of normal liver LO2 cells and hepatocellular carcinoma HepG2 cells. Methods: By using pAd-Easy-1 adenovirus vector system, the recombinant adenovirus Ad-SREBP2m (carrying the splicing form of SREBP2) and Ad-GFP (as the control) were constructed, and then infected into LO2 and HepG2 cells, respectively. The total cholesterol level in LO2 and HepG2 cells after infection was detected by a cholesterol quantification kit. The expression levels of SREBP2m, cholesterol synthesis rate-limiting enzyme 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR) and apoptosis-related proteins (including caspase 3, cleaved-caspase 3 and caspase 12) were detected by Western blotting. The effects of Ad-SREBP2m overexpression on the proliferation, cell cycle, apoptosis and migration of LO2 or HepG2 cells were detected by EdU staining method, FCM and scratch wound healing test, respectively. Results: The recombinant adenovirus Ad-SREBP2m was successfully constructed. Compared with the Ad-GFP group, the expression levels of SREBP2m and its target protein HMGCR were up-regulated (both P < 0.05), and the total cholesterol level increased significantly in LO2 cells and HepG2 cells infected with Ad-SREBP2m (both P < 0.05). In LO2 cells infected with Ad-SREBP2m, the proportion of G1-, S- and G2-phase cells was not significantly changed (all P > 0.05); while in HepG2 cells infected with Ad-SREBP2m, the proportion of G1-phase cells decreased significantly (P < 0.001), but the proportion of S-phase cells increased significantly (P < 0.001). SREBP2m overexpression promoted the proliferation of HepG2 cells (P < 0.001), but had no effect on the proliferation of LO2 cells. The expression levels of total caspase 3, cleaved-caspase 3 and caspase 12 were significantly higher in LO2 cells infected with Ad-SREBP2m than those in Ad-GFP group (all P < 0.001), while the expression levels of total caspase 3, cleaved-caspase 3 and caspase 12 were decreased in HepG2 cells infected with Ad-SREBP2m (all P < 0.05). The apoptosis rate of LO2 cells after Ad-SREBP2m infection was increased by (11.40±0.52)% (P < 0.001), while the apoptosis rate of HepG2 cells in Ad-SREBP2m group was decreased by (4.17±0.47)% as compared with Ad-GFP group (P < 0.05). The migration distance of HepG2 cells in Ad-SREBP2m group was (1.17±0.12) mm more than that in Ad-GFP group (P < 0.05). Conclusion: Overexpression of SREBP2m can promote the proliferation and migration and inhibit the apoptosis of hepatocellular carcinoma HepG2 cells, but it can promote apoptosis of normal liver LO2 cells. Copyright © 2018 by TUMOR. All rights reserved.     

TRPM8 promotes aggressiveness of breast cancer cells by regulating EMT via activating AKT/GSK-3β pathway

Breast cancer already taken the first place of incidence in Chinese female cancer patients. TRPM8 is found to be over-expressed in breast cancer, but whether it promotes breast cancer aggressiveness remains unknown. In our study, TRPM8 was identified highly expressing in all the tested breast cancer cell lines including MCF-7, T47D, MDA-MB-231, BT549, SKBR3 and ZR-75-30, while it just could be detected in MCF-10A, the normal breast epithelial cell. Then four pairs of clinical samples were analyzed using Western blotting and the result showed that TRPM8 expression is higher in tumor tissues than in adjacent nontumor tissues. Subsequently, we established TRPM8 high-expressing MCF-7 cell line and TRPM8 knockout MDA-MB-231 cell line to explore expression status of cancer-related proteins. The Western blotting and immunofluorescence analysis outcomes demonstrated that TRPM8 might influence cancer cell metastasis by regulating the EMT phenotype via activating AKT/GSK-3β pathway, and the hypothesis had been supported by cell function tests. All the results demonstrated that TRPM8 significantly up-expressed in breast cancer cells and promoted their metastasis by regulating EMT via activating AKT/GSK-3β pathway, indicating TRPM8 gets the prospects of to be developed as medication or diagnostic indicator to be applied in clinical work.     

Combination of microwave hyperthermia with epirubicin inhibits the proliferation and induces the apoptosis of breast cancer cells北大核心CSCD

Objective: To investigate the effects of microwave hyperthermia in combination with epirubicin on the growth of orthotopic breast transplantation tumor in BALB/c mice and the proliferation of murine breast cancer 4T1 cells and human breast cancer MDA-MB-231 cells, and to explore their possible mechanisms. Methods: The 4T1 cells were injected into mammary fat pad of female BALB/c mice to establish the orthotopic breast transplantation tumor model. The tumor-bearing mice were treated with microwave hyperthermia, epirubicin and microwave hyperthermia in combination with epirubicin, respectively. The breast tumor volume and weight and the number of lung metastatic nodes in mice were observed. The 4T1 and MDA-MB-231 cells were treated with microwave hyperthermia, epirubicin and microwave hyperthermia in combination with epirubicin, respectively. Then the cell proliferation and apoptosis were detected by MTS assay and FCM method, respectively. The change of phosphatidylinositol 3-kinase (PI3K)/mammalian target of rapamycin (mTOR) pathway and the expressions of apoptosis-related proteins were detected by Western blotting. Results: The tumor volume (P < 0.05) and weight (P < 0.05)and the number of lung metastatic nodes (P < 0.01) in mice in microwave hyperthermia in combination with epirubicin group were all smaller than those in the control group (the tumor-bearing mice didn't receive any treatment). The proliferation of breast cancer 4T1 and MDA-MB-231 cells in microwave hyperthermia in combination with epirubicin group was inhibited and the apoptosis was induced (all P < 0.05). Combination of epirubicin with microwave hyperthermia suppressed the activation of mTOR in 4T1 and MDA-MB-231 cells (both P < 0.05) and up-regulated the expressions of apoptosis-related proteins (all P < 0.05). Conclusion: Combination of microwave hyperthermia with epirubicin can suppress the growth and metastasis of breast cancer. This effect may be associated with the down-regulation of mTOR pathway and the up-regulation of apoptosis-related protein expressions. Copyright © 2018 by TUMOR. All rights reserved.     

Progress in the relationship between diabetes and prognosis of colorectal cancer北大核心CSCD

It has been found that diabetes is closely correlated with the occurrence of colorectal cancer, as an independent risk factor for colorectal cancer. In order to prolong the survival time and to improve the quality of life of colorectal cancer patients with diabetes, many studies have explored the influence of diabetes on the prognosis of patients with colorectal cancer in recent years. Some studies have confirmed diabetes is related to the prognosis of colorectal cancer, while other studies have suggested that diabetes is not correlated with the prognosis of colorectal cancer. Until now, it is still unclear whether the presence of diabetes has impact on the prognosis of colorectal cancer. Therefore, this problem has become one of the hot topics in cancer research field recently. This article reviews the research progress in the relationship between diabetes and the prognosis of patients with colorectal cancer. Copyright © 2017 by TUMOR All rights reserved.