Metastasis Of Colon Cancer Cells Research Articles

Let‑7i‑5p inhibits the proliferation and metastasis of colon cancer cells by targeting kallikrein‑related peptidase6.

Colorectal cancer is one of the most common types of malignancies worldwide, and as it is often diagnosed at an advanced stage, it is a serious threat to human health. MicroRNAs are important regulators of the growth and metastasis of colon cancer (CC). In the present study, the results demonstrated that kallikrein‑related peptidase6 (KLK6) plays a critical role in suppressing colon carcinoma progression. To further investigate whether microRNAs affect the impact of KLK6, a bioinformatics approach was employed, which indicated that let‑7i‑5p may directly target KLK6. Furthermore, the expression level of let‑7i‑5p was significantly negatively correlated with the expression of KLK6 at the mRNA and protein levels in CC. Functionally, overexpression of let‑7i‑5p inhibited the proliferation and invasion of CC cells, and suppressed the growth of CC invitro. The luciferase reporter assays revealed that let‑7i‑5p targeted the KLK6 3'‑untranslated region. Collectively, these results indicated that let‑7i‑5p inhibited the proliferation and metastasis of CC cells by targeting KLK6, thereby blocking the cell cycle and promoting apoptosis in colon cells. Therefore, the present study revealed that the let‑7i‑5p/KLK6 axis may be a potential target for new therapeutic strategies to treat colon tumors.

MicroRNA‑449a is a potential predictor of colitis‑associated colorectal cancer progression

An early diagnosis of colitis‑associated colorectal cancer (CAC) is important for its clinical management. However, it is currently difficult to distinguish the different stages of CAC development. MicroRNA dysregulation is common in human colorectal disorders, however little is known regarding whether miRNA affects tumor progression by regulating inflammation. In the present study, we identified a novel miRNA (miR‑449a), the expression of which was significantly reduced in CAC tissues than in paired adjacent non‑cancerous tissues (ANTs). Notably, the level of miR‑449a was in a markedly decreased pattern during the neoplastic transformation of ulcerative colitis (UC)‑to‑CAC, as demonstrated by both clinical investigations and the experimental mouse model induced by AOM/DSS treatment. In addition, we observed that decreased miR‑449a expression was associated with advanced T or Nstatus, later clinical stage and poor histological differentiation of CAC. Mechanistic studies revealed that miR‑449a inhibited the growth and metastasis of human colon cancer cells by directly binding to the 3'‑UTR of Notch‑1 and thereby, suppressed the activation of the Notch signaling pathway. Therefore, these findings provide strong evidence for the translational potential of miR‑449a in the discrimination of patients with UC that is likely to progress into CAC, from those unlikely to progress, as well as in the prognosis and diagnosis of CAC.

Abstract 1997: Pro-inflammatory cytokine IL-6 induces the upregulation and deacetylation of FRA-1, thus promoting colon cancer stemness and aggressiveness

Abstract Colon cancer is an aggressive epithelial tumor type encompassing cancer stem cells (CSCs) that not only contribute to its grow at the primary site but also to its malignant progression, resistance to chemotherapy, and distant metastases thus underlying poor patient outcomes. We have previously reported that significant upregulation of the gene encoding for the Fos-related antigen-1 (FOSL1) upon pro-inflammatory cytokine interleukin-6 (IL-6)-driven epithelial-to-mesenchymal transition (EMT) in colon cancer. However, the underlying molecular and cellular mechanism through which inflammatory cytokines like IL6 regulate CSCs plasticity during colon cancer onset and progression development remain unclear. Here, we show that IL-6 enhanced the stemness, drug resistance and metastasis of colon cancer cells by upregulation and deacetylation of the transcription factor FRA1. Ectopic expression of FRA1 in colon cancer cells induced stem-like properties and metastasis, while its knockdown suppressed these IL-6 induced effects. IL-6 enhanced FRA1 transcription through the STAT3 pathway. Also, we show that FRA1 directly interacts with the promoter of the CSC transcriptional factor Nanog and transactivates its expression to modulate colon cancer stemness. Furthermore, deacetylation of lysine 116 (K116) in FRA1 was essential for its activity on Nanog transcription. Preliminary analyses with Trichostatin A (TSA), an inhibitor of histone deacetylases (HDACs) I and II, but not nicotinamide (NAM), an inhibitor of the sirtuin (SIRT) family of deacetylases, showed that some member of the HDACs family might be responsible for FRA1 deacetylation. Finally, we examined the expression of IL6/STAT3-pY(705)/FRA1/k116ac/Nanog/HDACs/β-catenin in 120 paraffin-embedded human colon cancer tissues by immunohistochemistry (IHC). We found that while FRA1 was mainly localized in the nucleus of cancer cells, K116ac of FRA1 was present both in the nucleus and cytoplasm and showed a low level of Lys116 acetylation in the FRA1 nuclear fraction. Moreover, analysis of patient-derived colon cancer samples supported the clinical relevance of the IL-6/STAT3/FRA1/Nanog axis in contributing to the increased colon cancer stemness and metastasis. We propose that a transcriptional regulatory cross-talk between pro-inflammatory and cancer stemness signaling pathways drives the reprogramming of colon cancer cells to CSCs thus promoting progression towards malignancy and distant metastasis. Combinations of anti-IL6 therapies with HDACs inhibitors may offer a potential novel approach for colon cancer treatment. Keywords: Inflammation, cancer cell stemness, transcription, deacetylation, colon cancer Citation Format: Tingyang Wang, Jimin Shao. Pro-inflammatory cytokine IL-6 induces the upregulation and deacetylation of FRA-1, thus promoting colon cancer stemness and aggressiveness [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1997.

Abstract 3930: Chemoprevention of metastatic colon cancer by a novel small molecule

Abstract Introduction: Colon cancer is a one of the leading cause of death in both men and women. Metastatic colon cancer is responsible for mortality due to resistance to conventional therapies. Our group has previously reported that activation of AKT and Notch1 plays an important role in metastasis of colon cancer cells. As Notch1 and AKT regulates cell proliferation and are upstream of epithelial to mesenchymal transition (EMT) cascade, it seems interesting to explore their inhibition for pharmacological intervention. In the present study we identified a sesquiterpene molecule, Verrucarin J (VJ), that inhibited Notch1 expression and downregulated AKT mediated EMT transition in colon cancer cells. Methods: The anticancer effect of VJ was assessed on both colon cancer cells and stably AKT overexpressing colon cells by cell proliferation, apoptosis and Western blot analysis. For xenograft studies, pCMV/HCT-116 or AKT/HCT-116 cells (1.5 × 106) in a 50-μl final volume of phosphate-buffered saline (PBS) were injected subcutaneously into separate flanks of the mice. Also effects of VJ were determined in mouse model of colorectal cancer i.e. APC min+/_ mice. Statistical analysis was done by unpaired Student's t-test and one way ANOVA,*p≤0.05, **p≤0.01, ***p≤0.001. Results: Molecular analysis of VJ treatment on colon cancer cells revealed that it inhibited colon cancer growth by down regulating AKT and Notch1 signaling. Western blot analysis revealed that VJ inhibits AKT/NF-κβ/ Bcl2- signaling axis in colon cancer cells. VJ also Inhibited migration and invasion of colon cancer cells that corresponds with downregulation of mesenchymal marker expression. In addition, VJ treatment induces apoptosis in colon cancer cells as well as AKT overexpressing cells. Intraperitoneal administration of VJ in pCMV/HCT116 and AKT/HCT-116 xenograft mice revealed decrease in tumor volume in comparison with control mice. We also examined the in vivo efficacy of VJ in ApcMin/+ mouse model. Treatment of ApcMin/+ with VJ (0.5 mg/kg/IP/twice a week) over 12 weeks significantly reduced the number of intestinal polyps (distal 84%; Middle 63% and Proximal 2%) as compared vehicle treated mice. In addition, we also observed 50% reduction in number of colon tumors in VJ treated mice. Conclusion: In conclusion, our studies suggest that VJ inhibits both AKT and Notch1 signaling and induces apoptosis in colon cancer cells. In vivo studies revealed significant reduction in tumor formation in mice models of colon cancer. Hence VJ could be a viable therapeutic agent for treating patients with metastatic colon cancer. Citation Format: Deeksha Pal, Balaji Chandrasekaran, Ashish Tyagi, Becca Von Baby, Houda Alatassi, Murali Ankem, Chendil Damodaran. Chemoprevention of metastatic colon cancer by a novel small molecule [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3930.

LncRNA CASC15 promotes colon cancer cell proliferation and metastasis by regulating the miR‑4310/LGR5/Wnt/β‑catenin signaling pathway.

Previous studies have indicated that overexpression of long noncoding RNA cancer susceptibility 15 (CASC15) may promote tumor development and progression in gastric cancer and hepatocellular carcinoma. However, the function of CASC15 in colon cancer remains unknown. In the present study, the expression of CASC15 was upregulated in colon cancer tissues and its expression was correlated with clinical Tumor‑Node‑Metastasis stage and tumor metastasis. In addition, knockdown of CASC15 significantly inhibited the proliferation, migration and invasion of colon cancer cells invitro and invivo. Following mechanistic experiments, CASC15 was observed to act as a sponge to suppress microRNA (miR)‑4310 that targeted LGR5. Through the inhibition of miR‑4310, CASC15 promoted leucine‑rich repeat‑containing G‑protein coupled receptor 5 (LGR5) expression and consequently activated the Wnt/β‑catenin signaling pathway. The results revealed that the inhibition of the Wnt/β‑catenin signaling pathway in CASC15‑overexpressing colon cancer cells suppressed cellular proliferation, migration and invasion. Collectively, these results demonstrated that CASC15 promoted colon cancer growth and metastasis through the activation of the Wnt/β‑catenin signaling pathway in a miR‑4310/LGR5 dependent manner. Thus, the present study suggested that CASC15 may be a therapeutic target for colon cancer treatment.

Klotho negatively regulated aerobic glycolysis in colorectal cancer via ERK/HIF1\u03b1 axis

BackgroundKlotho (KL) was originally characterized as an aging suppressor gene, and has been identified as a tumor suppressor gene in a variety of cancers, including colorectal cancer. Recent years have witnessed the importance of metabolism transformation in cancer cell malignancies maintenance. Aberrant cancer cell metabolism is considered to be the hallmark of cancer. Our previous studies demonstrated that KL played negative roles in colon cancer cell proliferation and metastasis. However, its role in the cancer cell reprogramming has seldom been reported. The aim of this study was to examine the role of KL in aerobic glycolysis in colorectal cancer.MethodsCombining maximum standardized uptake value (SUVmax), which was obtained preoperatively via a PET/CT scan, with immunohistochemistry staining, we analyzed the correlation between SUVmax and KL expression in colorectal cancer tissues. The impact of KL on glucose metabolism and its mechanisms were further validated in vitro and in vivo.ResultsPatients with lower KL expression exhibited higher 18F-FDG uptake (P < 0.05), indicating that KL might participate in aerobic glycolysis regulation. In vitro assay by using colon cancer cell lines further supported this observation. By overexpressing KL in HTC116 and SW480 cells, we observed that the glycolysis was inhibited and the mitochondrial respiration increased, indicating that KL was a negative regulator of aerobic glycolysis. To seek for the underlying mechanisms, we tried to dig out the relation between KL and HIF1α signaling pathway, and found that KL negatively regulated HIF1α protein level and transcriptional activity. Western blot analysis showed that KL overexpression negatively regulated ERK pathway, and KL regulated aerobic glycolysis in part through its regulation of ERK/ HIF1α axis.ConclusionsTaken together, KL is a negative regulator of aerobic glycolysis and KL inhibited glucose metabolism transformation via the ERK/ HIF1α axis.

SERPINA3 Silencing Inhibits the Migration, Invasion, and Liver Metastasis of Colon Cancer Cells.

To investigate the impact of SERPINA3 on the migration, invasion, and liver metastasis of colon cancer cells. Immunohistochemical staining was conducted to determine SERPINA3 expression in the cancer and adjacent normal tissues of 131 patients suffering from colon cancer. In vitro experiment, colon cancer cells with low (HT-29P), intermediate (KM-12C), and high (HT-29LMM, KM-12L4) metastatic potential were obtained to examine SERPINA3 expression levels. Besides, quantitative real-time PCR (qRT-PCR) and Western Blot were performed to detect SERPINA3 expression in HT-29LMM and KM-12L4 cells transfected with SERPINA3 siRNA; Wound-healing and Transwell assays to measure cell migration and invasion, respectively; and ELISA to detect MMP-2 and MMP-9 levels. In vivo experiment, mice with liver metastasis of colon cancer were established to observe the effect of SERPINA3 silencing on liver metastasis. Immunohistochemical assay was applied to evaluate the expressions of Serpina3, Mmp-2, Mmp-9, and proliferating cell nuclear antigen (Pcna) in liver metastasis tissues. SERPINA3 in colon cancer tissues was higher than in adjacent normal tissues, which was associated with patients' clinicopathological features. Besides, SERPINA3 expression showed a rising trend in low, intermediate, and high metastatic potential colon cancer cells. After KM-12L4 and HT-29LMM cells transfected with SERPINA3 siRNA, the migration and invasive ability of cells, as well as the expression levels of MMP-2 and MMP-9 were all decreased. Moreover, SERPINA3 siRNA could not only reduce live metastasis of mice, but also down-regulate the expression of Mmp-2 and Mmp-9 in liver metastasis tissues. SERPINA3 silencing could inhibit the migration, invasion, and liver metastasis of colon cancer cells.

Targeting PRPK Function Blocks Colon Cancer Metastasis.

The biological functions of the p53-related protein kinase (PRPK) remain unclear. We have previously demonstrated that PRPK is phosphorylated by the T-LAK cell-originated protein kinase (TOPK) and that phosphorylated PRPK (p-PRPK) promotes colon cancer metastasis. Here, we analyzed colon adenocarcinomas from 87 patients and found that higher expression levels of p-PRPK were associated with later stages of metastatic dissemination (stage III and IV) as compared with earlier stages (stages I and II). Indeed, levels of p-PRPK were higher in metastatic versus malignant human colon adenocarcinomas. Knocking down PRPK expression attenuated colorectal liver and lung metastasis of colon cancer cells in vivo An in vitro kinase assay indicated that active PRPK does not phosphorylate p53 directly. We found that PRPK phosphorylates survivin, a regulator of colon cancer metastasis. PRPK phosphorylates survivin at Thr34, which is important for survivin stability. Taken together, our data strongly suggest that the PRPK signaling pathway promotes colon cancer metastasis by modulating survivin stability, and that PRPK could be a new prognostic marker for the survival of colon cancer patients. In addition, we identified an FDA-approved bacteriostatic antibiotic, fusidic acid sodium salt (fusidic acid or FA) as an inhibitor of PRPK, and show that FA combined with 5-fluorouracil (5-FU) inhibited PRPK activity and colon cancer metastasis to the lung in mice. We contend that the combination of FA with 5-FU could be an alternative therapeutic strategy to traditional chemotherapy for colon cancer patients with poor prognosis. Mol Cancer Ther; 17(5); 1101-13. ©2018 AACR.

Caspase-3 regulates the migration, invasion and metastasis of colon cancer cells.

Caspase-3 (CASP3) is a major mediator of apoptosis activated during cellular exposure to cytotoxic drugs, radiotherapy or immunotherapy. It is often used as a marker for efficacy of cancer therapy. However, recent reports indicate that caspase-3 has also non-apoptotic roles such as promotion of tumor relapse and tumor angiogenesis. Therefore, the roles of caspase-3 in tumor progression remain to be defined clearly. In our study, we established caspase-3 knockout (KO) colon cancer cell lines by use of the CRISPR technology. In vitro, caspase-3 knockout HCT116 cells were significantly less clonogenic in soft agar assays. They were also significantly less invasive and more sensitive to radiation and mitomycin C than control cells. In vivo, CASP3KO cells formed tumors at rates similar to control cells but were significantly more sensitive to radiotherapy. They were also less prone to pulmonary metastasis when inoculated either subcutaneously or intravenously. At the mechanistic level, caspase-3 gene knockout appeared to cause reduced EMT phenotypes when compared to parental HCT116 cells. Indeed, they showed significantly increased E-cadherin expression, reduced N-cadherin, Snail, Slug and ZEB1 expression than control cells. Therefore, therapeutic targeting of caspase-3 may not only increase the sensitivity of cancer cell to chemotherapy and radiotherapy, but also inhibit cancer cell invasion and metastasis.

Tetrandrine suppresses adhesion, migration and invasion of human colon cancer SW620 cells via inhibition of nuclear factor-κB, matrix metalloproteinase-2 and matrix metalloproteinase-9 signaling pathways

Tetrandrine (TET) exhibits biological activities, including anticancer activity. In Chinese medicine, TET has been used to treat hypertensive and arrhythmic conditions and has been demonstrated to induce cytotoxic effects on human cancer cell lines. However, to the best of the author's knowledge, no previous studies have revealed that TET affects cell metastasis in SW620 human colon cancer cells. The present study demonstrated that TET decreased the cell number and inhibited cell adhesion and mobility of SW620 cells. Furthermore, a wound healing assay was performed to demonstrate that TET suppressed cell movement, and Transwell chamber assays were used to reveal that TET suppressed the cell migration and invasion of SW620 cells. Western blotting demonstrated that TET significantly reduced protein expression levels of SOS Ras/Rac guanine nucleotide exchange factor 1, phosphatidylinositol 3-kinase, growth factor receptor bound protein 2, phosphorylated (p)-c Jun N-terminal kinase 1/2, p-p38, p38, 14-3-3, Rho A, β-catenin, nuclear factor-κB p65, signal transducer and activator of transcription-1 and cyclooxygenase-2, in comparison with untreated SW620 cells. Overall, the results of the present study suggested that TET may be used as a novel anti-metastasis agent for the treatment of human colon cancer in the future.

Effect of mitochondrial uncouplers niclosamide ethanolamine (NEN) and oxyclozanide on hepatic metastasis of colon cancer

Metabolism of cancer cells is characterized by aerobic glycolysis, or the Warburg effect. Aerobic glycolysis reduces pyruvate flux into mitochondria, preventing a complete oxidation of glucose and shunting glucose to anabolic pathways essential for cell proliferation. Here we tested a new strategy, mitochondrial uncoupling, for its potential of antagonizing the anabolic effect of aerobic glycolysis and for its potential anticancer activities. Mitochondrial uncoupling is a process that facilitates proton influx across the mitochondrial inner membrane without generating ATP, stimulating a futile cycle of acetyl- CoA oxidation. We tested two safe mitochondrial uncouplers, NEN (niclosamide ethanolamine) and oxyclozanide, on their metabolic effects and anti-cancer activities. We used metabolomic NMR to examine the effect of mitochondrial uncoupling on glucose metabolism in colon cancer MC38 cells. We further tested the anti-cancer effect of NEN and oxyclozanide in cultured cell models, APCmin/+ mouse model, and a metastatic colon cancer mouse model. Using a metabolomic NMR approach, we demonstrated that mitochondrial uncoupling promotes pyruvate influx to mitochondria and reduces various anabolic pathway activities. Moreover, mitochondrial uncoupling inhibits cell proliferation and reduces clonogenicity of cultured colon cancer cells. Furthermore, oral treatment with mitochondrial uncouplers reduces intestinal polyp formation in APCmin/+ mice, and diminishes hepatic metastasis of colon cancer cells transplanted intrasplenically. Our data highlight a unique approach for targeting cancer cell metabolism for cancer prevention and treatment, identified two prototype compounds, and shed light on the anti-cancer mechanism of niclosamide.

The Effect of Plasma Activated Verbascoside on HIF-1A Expression as Valuable Metastasis and Angiogenesis factor in HT-29 Colon Cancer Cells

Colon cancer is one of the most common and most prevalent Neoplasma in men; adenocarcinomas make up 95 percent of all colon cancer cases. Due to the side effects of conventional treatment methods such as chemotherapy, today, attentions have been attracted to natural compounds that have less side effects and anticancer properties. In attempting to investigate factors which may account for cancer treatment, this study raises two interrelated questions. Firstly, what is the effect of Verbascoside as a natural compound in inhibiting HIF-1α signaling pathway which is an effective route in the metastasis, angiogenesis and cell migration in the human cell line of HT-29 colon cancer; second, is it possible to enhance and raise the effect of this herbal drug by cold atmospheric pressure plasma jet? The focus of this study will be investigating the effect of Verbascoside as a natural compound in inhibiting HIF-1α signaling pathway and also examining the effect of atmospheric pressure plasma on the function of Verbascoside. A the first step, 29-HT cells were treated at different concentrations of verbascoside 10, 20, 30, 40, 50, 70, 100 μg / ml for 24 hours, then MTT test was used to calculate the inhibitory concentration. The colon cancer cell migration was evaluated by scratch test and HIF-1α gene expression was evaluated by Real Time PCR. The results of the MTT assay showed that verboscoside dose-dependent inhibition of cancer cells. The HT-29 colon is at a concentration of 50 μg/ml. Scratch repair tests showed that verboscoside reduced the migration of cells in a dose-dependent manner, and the results of Real Time PCR showed a decrease in the expression of HIF-1α gene expression. Cancer tumors undergo some changes in oxygen deficiency conditions, including increasing the expression of the HIF-1α gene, which in turn play an important role in activating and enhancing the expression of genes involved in metastasis, angiogenesis and migration. At the second step of this study, verboscoside was primarily treated by cold atmospheric plasma jet then the same procedure has been taken for plasma activated verboscoside. In this study helium jet has been used.

Overexpression of Sirt6 is a novel biomarker of malignant human colon carcinoma.

Sirt family has been reported playing a significant role in the cancer development, especial its deacetylase activity plays a key function, but whether SIRT6 plays a role in mediating tumor epithelial-mesenchymal transition (EMT) and metastasis in colon cancer has not been explored. Here, the mass spectrometry and co-immunoprecipitation assays were utilized to detect that SIRT6 was physically associated with transcription factor snail. Most important, HCT116 cells transfected with SIRT6 shRNA reversed EMT, while increased the expression of TET1, and the HCT116 cells transfected with SIRT6 displayed the contrary tendency. Transwell invasion assay, soft agar assay, as well as colony formation together showed that SIRT6 promoted cell EMT and tumorigenesis in vitro. We also found SIRT6 is a reader of snail. ChIP as well as qChIP suggested H3K9 binding on the promoter of TET1 dependent on SIRT6. SIRT6 promoted EMT process through two different ways, one is as a reader of snail, and other way was the suppression of TET1 transcription. These two ways are all dependent on its H3K9 deacetylase activity. Further, patient samples collected showed that SIRT6 was significantly increased in colon cancer samples, and its higher expression was correlated with poor prognosis, worse overall survivals. Together, our experiments revealed the mechanism for SIRT6 in facilitating tumorigenesis and metastasis of colon cancer cells, suggesting that SIRT6 might be a potential therapeutic target for treating colon cancer.

Aspirin inhibited the metastasis of colon cancer cells by inhibiting the expression of toll-like receptor 4

BackgroundThe metastasis of colorectal cancer frequently tends to liver, which is one of the three leading causes of cancer-related deaths worldwide. Growing evidence showed that aspirin could effectively inhibit liver metastasis of colorectal cancer. However, the potential mechanism has not been fully understood.MethodsMouse splenic vein metastasis assay was used to examine the metastatic ability of colon cancer cells in vivo. And wound healing and transwell assay were applied to detect the metastasis potential of C26 and HCT116 colon cancer cell lines in vitro. RT-PCR and western blotting were used to explore Toll-like receptor 4 (TLR4) expression in colon cancer cell lines. The functions of TLR4 in the migration of the colon cancer cell line were analyzed by infecting cells with lentivirus containing TLR4 siRNA.ResultsWe demonstrated that lipopolysaccharides (LPS) could enhance the metastasis potential of C26 and HCT116 colon cancer cell lines. However, aspirin effectively decreased the metastasis capacity of colon cancer cells in vitro and in vivo. We found that the enhancement of LPS on the migration of colon cancer cells by inducing epithelial-mesenchymal transition (EMT) phenotype demonstrated a TLR4-dependent manner. Aspirin treatment lead to the downregulation of TLR4 on C26 cells which resulted in the decrease of C26 cells migration and EMT phenotype that induced by LPS. Additionally, the inhibitory effect from aspirin on the expression of TLR4 on C26 cells leads to the downregulation of NF-κB.ConclusionThe results of our study indicate that LPS origin from intestinal flora may promote the metastasis of colon cancer to liver and aspirin may inhibit the metastasis of colon cancer by inhibiting the expression of TLR4.

Mechanism of chlorogenic acid in apoptotic regulation through AKT/GSK-3β pathway in colon cancer cell

Objective To investigate the regulation mechanism of chlorogenic acid (CGA) on proliferation and apoptosis of colon cancer cell. Methods The effect of CGA with different concentrations on the apoptosis of human colon cancer cell line HT-29 was detected by flow cytometry, the cell proliferation was detected by MTT assay, and the cell migration was detected by Transwell method. The nude mice tumor model of HT-29 cells was established by injecting 100 μl CGA (100 μg/ml) into the tumor area, and 100 μl saline was injected into the same area of control group. The tumor volume and quality were measured periodically. The expression level of AKT/GSK-3β signaling pathway was detected by Western Blot. Results CGA can significantly promote the apoptosis of HT-29 cells (P 0.05). The phosphorylated proteins (p-AKT and p-GSK-3) and β-catenin in the experimental group were significantly lower than those in the control group (all P<0.01). Conclusion CGA can inhibit the proliferation and metastasis of colon cancer cells, and promote apoptosis. This mechanism might involve AKT/GSK-3β pathways. Key words: Colon cancer; Chlorogenic acid; AKT/GSK-3β pathways; Proliferation; Apoptosis

Phthalate exposure promotes chemotherapeutic drug resistance in colon cancer cells.

Phthalates are widely used as plasticizers. Humans can be exposed to phthalates through ingestion, inhalation, or treatments that release di(2-ethylhexyl) phthalate (DEHP) and its metabolite, mono(2-ehylhexyl) phthalate (MEHP), into the body from polyvinyl chloride-based medical devices. Phthalate exposure may induce reproductive toxicity, liver damage, and carcinogenesis in humans. This study found that colon cancer cells exposed to DEHP or MEHP exhibited increased cell viability and increased levels of P-glycoprotein, CD133, Bcl-2, Akt, ERK, GSK3β, and β-catenin when treated with oxaliplatin or irinotecan, as compared to control. The P-glycoprotein inhibitor, tariquidar, which blocks drug efflux, reduced the viability of DEHP- or MEHP-treated, anti-cancer drug-challenged cells. DEHP or MEHP treatment also induced colon cancer cell migration and epithelial-mesenchymal transformation. Elevated stemness-related protein levels (β-catenin, Oct4, Sox2, and Nanog) and increased cell sphere sizes indicated that DEHP- or MEHP-treated cells were capable of self-renewal. We also found that serum DEHP concentrations were positively correlated with cancer recurrence. These results suggest phthalate exposure enhances colon cancer cell metastasis and chemotherapeutic drug resistance by increasing cancer cell stemness, and that P-glycoprotein inhibitors might improve outcomes for advanced or drug-resistant colon cancer patients.

CAFFEIC ACID PHENETHYL ESTER (CAPE) MEDIATED DECREASE IN METASTASIS OF COLON CANCER CELLS: AN IN VITRO AND IN VIVO STUDY

Background: Caffeic acid phenethyl ester (CAPE) is a phytochemically active component obtained from honeybee hive&#x0D; propolis. CAPE has been reported to show antimitogenic, anticancer, and other beneficial medicinal properties. Many of its&#x0D; activities have been reported to be mediated by inhibiting levels of matrix metalloproteinase, that is, MMP-2 and MMP-9.&#x0D; We hypothesize the effect of CAPE on the metastasis of colon cancer cells in both in vitro and in vivo.&#x0D; Methods: Cell migration, motility, invasion were evaluated also expression of protein and matrix metalloproteinases&#x0D; (MMPs) such as MMP-2 and MMP-9 were measured in SW-480 cancer cells in vitro. The cells were exposed to Phorbol&#x0D; 12-myristate 13-acetate (PMA) and were treated with various concentration of CAPE.&#x0D; Results: The treatment of CAPE caused significant decrease (P

Secretory RAB GTPase 3C modulates IL6-STAT3 pathway to promote colon cancer metastasis and is associated with poor prognosis

BackgroundRAB GTPases are important in the regulation of membrane trafficking and cell movement. Recently, exocytic RABs have received increasing attention in cancer research. However, the functional roles of exocytic RABs in colorectal carcinogenesis remain to be elucidated.MethodsImmunohistochemistry analysis of a microarray containing 215 colorectal adenocarcinoma tissues was used to identify the association between exocytic RABs and patient prognosis. Complementary functional RAB3C overexpression and knockdown experiments were performed. The molecular mechanism of RAB3C in inducing colon cancer cell metastasis was determined.ResultsHigh RAB3C expression in patients was found to be significantly associated with advanced pathological stage, distant metastasis and poor prognosis. Multivariate analyses showed that high RAB3C expression was an independent prognostic marker in overall (P = 0.001) and disease-free survival (P < 0.001). Furthermore, our experimental results showed an increase in the migration and invasion ability of RAB3C-overexpressing colon cancer cells and increased metastatic nodules in a mouse metastasis model. The effect of RAB3C-overexpressing cell-conditioned medium was found to significantly promote the migration ability of parental colon cancer cells, thus suggesting that the promotion of migration is exocytosis dependent. Upregulation of other exocytic RABs was also seen in RAB3C-overexpressing cells. Through microarray and proteomics analyses, increased production of multiple cytokines was observed in RAB3C-overexpressing cell lines, and the IL-6 pathway was the top pathway whose members exhibited gene expression changes after RAB3C overexpression, according to Ingenuity Pathway Analysis. Blocking IL-6 with IL-6 antibody treatment or IL-6 knockdown significantly inhibited the migration potential of RAB3C-overexpressing colon cancer cells. In addition, IL-6 was found to induce STAT3 phosphorylation in RAB3C-overexpressing colon cancer cells, thus promoting migration. Ruxolitinib, a JAK2 inhibitor, was found to significantly inhibit RAB3C-induced colon cancer cell migration.ConclusionsOur study revealed that RAB3C overexpression promotes tumor metastasis and is associated with poor prognosis in colorectal cancer, through modulating the ability of cancer cells to release IL-6 through exocytosis and activate the JAK2-STAT3 signaling pathway. These results further suggest that inhibition of STAT3 phosphorylation in the RAB3C-IL-6-STAT3 axis by using Ruxolitinib may be a new therapeutic strategy to combat metastatic colon cancers.

MiR-223 promotes colon cancer by directly targeting p120 catenin

microRNA (miRNA) dysregulation is frequently observed in colon cancer. Previous studies found that miR-223 is upregulated in colon cancer and functions as an oncogene. Conversely, p120 is often downregulated or even absent in colon cancer, and is a likely tumor suppressor. The present study showed that increased miR-223 and decreased p120 levels are associated with colon cancer malignancy, and p120 expression is negatively correlated with miR-223 expression. A dual luciferase reporter assay showed that miR-223 directly targets p120. miR-223 upregulation in a colon cancer cell line upregulated c-Myc, cyclinD1, MMP7, and vimentin expression, downregulated E-cadherin, increased nuclear expression of β-catenin, and enhanced RhoA activation. We suggest miR-223 may promote colon cancer cell invasion and metastasis by downregulating p120, thereby reducing intercellular adhesion, promoting RhoA activity, and activating β-catenin signaling. Thus miR-223 functions as an oncogene in colon cancer and may be a potential diagnostic and therapeutic target for anti-colon cancer treatment.

Expression of CXC chemokine ligand 16/CXC chemokine receptor 6 in colon cancer and its effect on proliferation, invasion and metastasis of colon cancer

Objective To investigate the expression of CXC chemokine ligand 16 (CXCL16)/CXC chemokine receptor 6 (CXCR6) in colon cancer and its effect on the proliferation, invasion and metastasis of colon cancer. Methods Immunohistochemistry was used to detect the expression of CXCL16/CXCR6 in colorectal cancer tissues and para carcinoma tissues, and colon cancer cell lines: SW480, LoVo, HCT116. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the expression of CXCL16 mRNA and CXCR6 mRNA. The methyl thiazol tetrazolium (MTT) and Transwell were used to analyze the effects of CXCL16/CXCR6 on the proliferation, invasion and metastasis of colon cancer cells by CXCR16-RNA lentiviral vector transfected SW480 cells. Results In 56 cases of colon cancer, there were 48 cases positive for CXCL12 expression, and 47 positive for CXCR6 expression, with the positive expression rate being 85.7% and 83.9% respectively, significantly higher than in the adjacent normal tissues (26.7%, 33.3%), the difference was statistically significant (χ2=18.542, P=0.005; χ2=19.464, P=0.005). CXCL16 and CXCR6 in colon cancer cell lines SW480, LoVo, HCT116 were all positively expressed. The RT-PCR revealed that the expression of CXCL16 and CXCR6 mRNA in cancerous tissues was statistically significantly higher than in the normal tissues adjacent to carcinoma (t=2.592, P=0.008; t=2.364, P=0.003). CXCL16 and CXCR6 mRNA was highly expressed in the cancer cell linves (P=0.232). After transfection with lentivirus vector, MTT experiment showed that the proliferation of CXCL16-short hairpin RNA (shRNA) group was significantly lower than the control group (P=0.002). Transwell experimental results demonstrated that the number of cells in the experimental group and the control group was (14.2±1.68)×105 and (28.7±2.02)×105 respectively, and the difference was statistically significant (t=3.022, P=0.001). Conclusion CXCL16/CXCR6 may be involved in the proliferation, invasion and metastasis of colon cancer. Key words: CXC chemokine ligand 16; CXC chemokine receptor 6; Colon cancer; Proliferation; Invasion