Inhibits Colon Cancer Cell Research Articles

MiR‑4262 inhibits colon cancer cell proliferation via targeting of GALNT4.

MicroRNAs (miRs) have been demonstrated to be important in the establishment and progression of colon cancer. However the underlying molecular mechanisms remain to be fully elucidated. Polypeptide N-acetylgalactosaminyltransferase4 (GALNT4) participates in numerous cellular processes, including tumorigenesis. The present study used reverse transcription-quantitative polymerase chain reaction and western blotting to investigate the expression levels of miR-4262 and GALNT4 in tissues and cells. In addition, MTS and colony formation assays, and cell cycle analysis were performed to evaluate the effect of miR-4262 on cell proliferation and the cell cycle. The findings demonstrated that miR-4262 was a direct target of GALNT4 mRNA. Overexpression of miR-4262 was demonstrated to decrease GALNT4 mRNA and protein expression levels, and thereby suppressed cell viability, growth and cell-cycle progression in SW480 and SW620 colon cancer cells. In addition, knockdown of miR-4262 significantly increased the cell viability, growth, and cell-cycle progression of SW480 and SW620 cells. The expression level of miR-4262 was observed to be downregulated as the expression of GALNT4 was upregulated in colon cancer tissues and cell lines. In conclusion, the results demonstrated that miR-4262 may be involved in the development of colon cancer via targeting of GALNT4. The miR-4262/GALNT4 axis may be a novel target for diagnosing and understanding the underlying molecular mechanism of colon cancer.

MicroRNA-136 inhibits colon cancer cell proliferation and invasion through targeting liver receptor homolog-1/Wnt signaling

An increasing number of studies have reported that microRNAs (miRNAs) are involved in the malignant behavior of colon cancer cells through directly targeting multiple tumor suppressors or oncogenes. The expression and role of miR-136 has been reported in several types of human cancer. However, the role of miR-136 in colon cancer remains unclear. In this study, we aimed to investigate the expression and function of miR-136 in colon cancer and the potential underlying mechanism. Here, we found that miR-136 was decreased in colon cancer cell lines and tissues. Overexpression of miR-136 inhibited the proliferation and invasion in SW480 and HCT116 cell lines while suppression of miR-136 exhibited the opposite effect. Liver receptor homolog-1 (LRH-1) was identified as a direct target gene of miR-136. Notably, miR-136 overexpression suppressed LRH-1 expression as well as Wnt signaling in SW480 and HCT116 cell lines. The miR-136 expression level inversely correlated with LRH-1 mRNA expression in colon cancer specimens. Moreover, overexpression of LRH-1 partially reversed the miR-136-induced antitumor effect in SW480 and HCT116 cell lines. Taken together, these findings suggest that miR-136 functions as a negative regulator in colon cancer progression by targeting LRH-1 and that miR-136 downregulation contributes to high expression of LRH-1 and aberrant activation of Wnt signaling, leaving open the possibility that miR-136 may serve as a potential therapeutic target for colon cancer.

Partially Purified Gloriosa superba Peptides Inhibit Colon Cancer Cell Viability by Inducing Apoptosis Through p53 Upregulation

BackgroundColon cancer is a major health problem worldwide. Available treatments such as surgery, chemotherapy, radiation and anticancer drugs are limited due to stage of cancer, side effects and altered biodistribution. The use of peptides extracted from natural products has appeared as a potential therapy. Gloriosa superba is known to contain colchicine and other alkaloids with anticancer activity. However, these peptides contained within the extracts have not been studied. This study, therefore, focuses on an investigation of anti–colon cancer activity from a partially purified protein hydrolysate of G superba rhizome. MethodsDried G superba rhizome was extracted using 0.5% sodium dodecyl sulfate and digested with pepsin. The protein hydrolysates with molecular weight lesser than 3kDa were collected and subjected for cell viability assay. Then, the partial purification of the protein hydrolysate was performed using reverse-phase high-performance liquid chromatography. Fractions containing anticancer peptides were investigated, and their effects on apoptosis and protein expression using apoptosis test and Western blot, respectively. ResultsPartially purified peptides of G superba rhizome demonstrated anticolon activity in SW620 cells by inducing apoptosis through upregulation of p53 and downregulation of nuclear factor kappa B (NF-κB). ConclusionsConsequently, G superba peptides showed high potential for further purification and development of anticolon therapeutics.

MicroRNA-1299 is a negative regulator of STAT3 in colon cancer.

Signal transducers and activators of transcription (STAT) is a family of transcription factors which regulate cell proliferation, differentiation, apoptosis, metastasis, immune and inflammatory responses, and angiogenesis. STAT3 is a latent cytoplasmic transcription factor that belongs to STATs. STAT3 has been reported be regulates genes involved with cellular growth, proliferation and metastasis. Worldwide, colon cancer is one of the leading causes of cancer-related deaths. Cumulative evidence has established that STAT3 is essential for colon cancer progression to advanced malignancy. In our study, we showed that microRNA-1299 (miR-1299) was closely related to the TNM stage of colon cancer, and that the expression of miR-1299 was negatively correlated with the expression of STAT3 in colon cancer which means that miR-1299 can be a negative regulator of STAT3 in colon cancer. A total of 60 cases of different grades of colon samples were used to detect the expression of miR-1299. Results showed that miR-1299 was significantly lower in high-grade colons both in mRNA and protein levels. Furthermore, Overall survival (OS) in patients with low miR-1299 is shorter than 25.6 months, as compared with an OS of 28.4 months in patients with high level of miR-1299. We also confirmed that the overexpression of miR-1299 can not only downregulate the STAT3 pathway, but also inhibited colon cancer cell growth. Our findings could provide new insights into the molecular therapeutic of colon cancer.

Crocetin treatment inhibits proliferation of colon cancer cells through down-regulation of genes involved in the inflammation.

BackgroundThe current study was designed to investigate the effect of crocetin on the proliferation inhibition of colon cancer cells and the underlying mechanism.MethodsMTT assay showed inhibition of proliferation of colon cancer cells in a dose based manner by crocetin treatment. At 30 µM concentration of crocetin proliferation rate of colon cancer cells was reduced to 14% after 24 h. Flow cytometry and fluorescence microscopy revealed induction of apoptosis in colon cancer cells on treatment with crocetin. The tube formation was suppressed significantly in the cultures of HUVEC treated with 30 µM concentration of crocetin compared to the control cultures.ResultsThe results from transwell assay revealed a significant reduction in the population of DU-145 cells passing through filters of transwell on treatment with crocetin compared to the control cells. Treatment of the DU-145 cells with crocetin caused a significant reduction in the expression levels of NF-κB, VEGF and MMP-9. The results from RT-PCR analysis revealed a significant reduction in the expression of genes involved in inflammation including, HMGB1, IL-6 and IL-8 on treatment of DU-145 cells with crocetin. However, the expression of NAG-1 gene was increased by crocetin treatment in DU-145 cells significantly compared to the control cells.ConclusionCrocetin inhibits growth of colon cancer cells and prevents tube formation through induction of apoptosis. Therefore, crocetin can be used efficiently for the treatment of colon cancer.

NCAPH plays important roles in human colon cancer

Colon cancer (CC) is one of the major malignancies worldwide, whose pathogenesis is complex and requires the accumulated alteration of multiple genes and signaling pathways. Condensins are multi-protein complexes that play pivotal roles in chromosome assembly and segregation during mitosis, meiosis and even tumorigenesis. Using tissue microarrays by immunohistochemistry and hematoxylin–eosin staining, we found that non-SMC condensin I complex subunit H (NCAPH) in colon cancerous tissues was higher than that in all corresponding adjacent non-cancerous tissues. We then characterized the exact function of the NCAPH in CC. We provided evidences showing that NCAPH is highly expressed in colorectal cancer cell lines comparing with normal human colonic epithelial cells, and identified many NCAPH mutations in CC patients. We found that depletion of NCAPH inhibits CC cell proliferation, migration in vitro and xenograft tumor formation in vivo. Furthermore, NCAPH knockdown promotes cell apoptosis and cell cycle arrest at G2/M phase. Interestingly, the NCAPH high expression in tumor tissues of colon patients had a significantly better prognosis and survival rate than low-expression patients, suggesting that NCAPH high expression promotes colonic cancerous cell proliferation; on the other hand, it may also sensitize these cells responding to chemo- or radio-therapies. Collectively, these findings reveal an important role of NCAPH in CC, indicating that NCAPH could be used as a new therapeutic target in future.

Dihydroartemisinin inhibits colon cancer cell viability by inducing apoptosis through up-regulation of PPARγ expression.

The present study was aimed to investigate the effect of dihydroartemisinin on the colon cancer cell proliferation and apoptosis. The results from MTT assay revealed a concentration and time dependent relation between the inhibition of SW 948 cell viability and dihydroartemisinin addition. The viability of SW 948 cells was reduced to 45 and 24% on treatment with 30 and 50 µM, respectively concentrations of dihydroartemisinin after 48 h. Morphological examination of SW 948 cells showed attainment of rounded shape and cluster formation on treatment with dihydroartemisinin. Western blot analysis showed a significant increase in the activation of caspase-3 and expression of cleaved PARP by dihydroartemisinin treatment. The activation of PPARγ was increased significantly in SW 948 cells by treatment with dihydroartemisinin. Compared to control, the migration potential of SW 948 cells was reduced significantly (p < 0.005) and the expression levels of MMP-2 and -9 inhibited by dihydroartemisinin at 50 µM concentration. In the dihydroartemisinin treatment group colon tumor formation was significantly inhibited on treatment with 20 mg/kg doses of dihydroartemisinin after 30 days. Therefore, dihydroartemisinin inhibits colon cancer growth by inducing apoptosis and increasing the expression of PPARγ. Thus dihydroartemisinin can be used for the treatment of colon cancer.

Benzoflavonoids Structure Hinders Human Colon Cancer Clonogenicity.

Since flavonoids fused by benzene have been known for their potent chemopreventive effects, in this study, we examined the relationship between the structures and activities of benzoflavones, benzoflavanones, benzochalcones, and benzochalcone derivatives bearing the pyrazole moiety against human colon cancer cells. We investigated the effect of 34 benzoflavonoids on the inhibition of colon cancer cells based on the clonogenicity. The biological activity values used for the quantitative structure-activity relationship (QSAR) calculations were obtained from the cell growth inhibition on the basis of clonogenicity. 3D-QSAR calculations were performed using comparative molecular field analyses (CoMFA) and comparative molecular similarity index analyses (CoMSIA). Of several CoMFA and CoMSIA models, the best models showing the highest cross validated correlation coefficient were selected and validated. The cell growth inhibition values were calculated using the above models. The structural conditions to show good cell growth inhibitory effects on human colon cancer cells were analyzed by CoMFA and CoMSIA contour maps. The contribution of steric fields remarkably decreased without any change in the contribution of the electrostatic field, which means that electrostatic contribution is more crucial than the steric contribution in the modification of benzoflavonoids. Furthermore, the increase in the hydrogen bond donor contribution was approximately proportional to the decrease in steric field contribution. This study demonstrated that benzoflavonoids structure hinders colon cancer clonogenicity. Most of the benzoflavonoids structures comprised a C-3 linkage between the naphthalene and phenyl moieties, which contained diverse functional moieties such as oxygen-fused rings, double bonds, pyrazole rings, and sulfur constituents, and were able to exhibit great potential in diverse anticancer effects. Also, the positions of the hydroxyl group close to the naphthalene and phenyl rings were crucial for activity against colon cancer. The structural conditions obtained here may help us design potent benzoflavonoids against colon cancer cells and predict their activities.

S-Allylmercaptocysteine induces G2/M phase arrest and apoptosis via ROS-mediated p38 and JNK signaling pathway in human colon cancer cells in vitro and in vivo

SAMC inhibits colon cancer cell growth through the reactive oxygen species-dependent pathway.

FH535 Inhibits Proliferation and Motility of Colon Cancer Cells by Targeting Wnt/β-catenin Signaling Pathway.

Aberrant Wnt/β-catenin pathway activation is frequently observed in human colorectal cancer (CRC) and has become a promising target for CRC treatment. Our study aimed to evaluate the effect of FH535, a small molecule inhibitor of Wnt/β-catenin pathway, on two colon cancer cell lines, HT29 and SW480. We found FH535 significantly inhibited colon cancer cell proliferation in vitro and induced cell cycle arrest. Moreover, FH535 inhibited colon cancer xenograft growth in vivo. Wound-healing assay and Transwell assay revealed that FH535 notably suppressed migration and invasion of SW480 cells. FH535 also repressed expression of cancer stem cell markers, CD24, CD44 and CD133 in HT29 cells. Real time-quantitative PCR and Western blotting revealed that targeting Wnt/β-catenin pathway using FH535 effectively downregulated target genes including cyclin D1 and survivin at mRNA and protein level, which contributed to the FH535-induced inhibitory effect on colon cancer cell proliferation. As mechanisms for suppressing cancer cell motility, FH535 downregulated expression of matrix metalloproteinase-7 and -9, Snail and vimentin. RNA sequencing revealed that FH535 prominently altered multiple biological pathways associated with DNA replication, cell cycle and metabolism. Our study highlights the anti-cancer effect of FH535 on colon cancer and presents its potential in colon cancer treatment.

Anti-cancer Effects of MW-03, a Novel Indole Compound, by Inducing 15-Hydroxyprostaglandin Dehydrogenase and Cellular Growth Inhibition in the LS174T Human Colon Cancer Cell Line.

Increases in the expression of prostaglandin E2 (PGE2) are widely known to be involved in aberrant growth in the early stage of colon cancer development. We herein demonstrated that the novel indole compound MW-03 reduced PGE2-induced cAMP formation by catalization to an inactive metabolite by inducing 15-hydroxyprostaglandin dehydrogenase through the activation of peroxisome proliferator-activated receptor-γ. MW-03 also inhibited colon cancer cell growth by arresting the cell cycle at the S phase. Although the target of MW-03 for cell cycle inhibition has not yet been identified, these dual anti-cancer effects of MW-03 itself and/or its leading compound(s) on colon cancer cells may reduce colon cancer development and, thus, have potential as a novel treatment for the early stage of this disease.

Four microRNAs Signature for Survival Prognosis in Colon Cancer using TCGA Data.

This study aims to develop microRNA expression signature for colon cancer survival prognosis based on the Cancer Genomic Common database. miRNAs levels between colon cancer and non-cancer tissues were screened by t-test (p < 0.05). Kaplan-Meier survival method was used to discriminate survival significant miRNAs, followed by miRNAs index accumulation to power the miRNAs-survival reliability. In the end, we test the selected miRNAs in HT126 colon cancer cells to validate its anti-cancer effect. The study identified a 84-miRNAs signature. Of the above 84 miRNAs, we got four miRNAs which were survival associated by using ROC curve method and Kaplan-Meier survival method (p < 0.001). The result showed that low risk group had quite a low death rate, the survival rate was over 80%. The high risk group had survival rate lower than 20%, which was also extremely lower than the overall survival rate. In the HT126 cells study, cell growth assay showed miR-130a sponge inhibited colon cancer cells growth and sensitized the anti-cancer drug effect of 5-FU to blocked cancer cell growth. We developed a prognostic 4-microRNA expression signature for colon cancer patient survival, and validated miR-130a sponge could sensitized 5-FU anti-cancer effect.

Preparation and characterization of polymeric nanoparticles surface modified with chitosan for target treatment of colorectal cancer

5-Fluorouracil (5-FU) loaded chitosan (C) coated polylactic-co-glycolic acid (PLGA) nanoparticles [NPs] (C-5-FU PLGA NPs) and polycaprolactone [PCL] (C-5-FU PCL NPs) were employed as the carriers for cancer treatment. The prepared NPs showed the spherical shape of NPs with the particle size in the range of 188.1–302.2nm with polydispersity index (PDI) of <0.30. C-coated NPs converted zeta potential from negative to positive value with small modification in particle size distribution. The entrapment efficiency of 5-FU was recorded in the range of 32–51%. The in vitro release studies showed an initial rapid 5-FU release followed by a sustained release profile. The in vitro cytotoxicity of C-5-FU PLGA NPs showed significant inhibition of colon cancer cells (HT-29) compared to the other NPs and drug solution. These results showed that C-5-FU PLGA NPs can be considered as a promising carrier for cancer therapy.

8‐C‐(E‐phenylethenyl)quercetin from onion/beef soup induces autophagic cell death in colon cancer cells through ERK activation

Quercetin, a flavonoid, widely distributed in edible fruits and vegetables, was reported to effectively inhibit 2-amino-1-methyl-6-phenylimidazo[4, 5-b]pyridine (PhIP) formation in a food model (roast beef patties) with itself being converted into a novel compound 8-C-(E-phenylethenyl)quercetin (8-CEPQ). Here we investigated whether 8-CEPQ could be formed in a real food system, and tested its anticancer activity in human colon cancer cell lines. LC-MS was applied for the determination of 8-CEPQ formation in onion/beef soup. Anticancer activity of 8-CEPQ was evaluated by using cell viability assay and flow cytometry. Results showed that 8-CEPQ suppressed proliferationand caused G2 phase arrest in colon cancer cells. Based on immunofluorescent staining assay, western blot assay, and RNA knockdown data, we found that 8-CEPQ did not cause apoptotic cell death. Instead, it induced autophagic cell death. Moreover, treatment with 8-CEPQ induced phosphorylation of extracellular signal-regulated kinase (ERK). Inhibition of ERK phosphorylation by the mitogen-activated protein kinasekinase (MEK)/ERK inhibitor U0126 attenuated 8-CEPQ-induced autophagy and reversed 8-CEPQ-mediated cell growth inhibition. Our results demonstrate that 8-CEPQ, a novel quercetin derivative, could be formed in onion/beef soup. 8-CEPQ inhibited colon cancer cell growth by inducing autophagic cell death through ERK activation.

Effects and Mechanism of Imatinib in Inhibiting Colon Cancer Cell Proliferation.

BackgroundThis study investigated the effects and mechanism of imatinib in inhibiting colon cancer cell proliferation.Material/MethodsThe SW480 cells were divided into 4 imatinib-treated groups: 0 μM, 1.25 μM, 2.5 μM, and 5μM. We analyzed the apoptosis and cell cycle of the 4 groups. The gene and protein expressions of p21, p27, HGF, and GAPDH were measured by RT-PCR and Western blot.ResultsCompared with the 0-μM imatinib-treated group, the apoptosis of 1.25-μM, 2.5-μM, and 5.0-μM treated groups was significantly induced (P<0.05, all). The G1 phase was significantly up-regulated in the 1.25-μM, 2.5-μM, and 5.0-μM treated groups compared with the 0-μM imatinib-treated group (P<0.05, respectively), but the S and G2 phase of 3 imatinib-treated groups were significantly down-regulated (P<0.05, all). The gene and protein expressions of p27 and HGF were significantly different among the 4 groups (P<0.05, all).ConclusionsImatinib inhibits proliferation of colon cancer cells by reducing HGF and increasing p27 in a dose-dependent manner.

Stereoisomers of Astaxanthin Inhibit Human Colon Cancer Cell Growth by Inducing G2/M Cell Cycle Arrest and Apoptosis.

Astaxanthin (AST) is a xanthophyll carotenoid with potential protective effects against carcinogenesis. Different stereoisomers of AST (ASTs) exist in a variety of food sources. Due to limited information on the bioactivities of ASTs, the present study investigated the inhibitory effects of ASTs on HCT116 and HT29 human colon cancer cells. ASTs investigated herein included 3S,3'S (S) from Haematococcus pluvialis, 3R,3'R (R) from Phaffia rhodozyma, and a statistical mixture (S: meso: R = 1:2:1) (M) from synthetic AST. Cell viability assay showed that ASTs all inhibited colon cancer cell growth in a time-dependent (24-72 h) and dose-dependent (4-16 μM) manner, and there was no significant difference among the IC50 values of ASTs (p > 0.05). Flow cytometry analysis indicated that ASTs induced G2/M cell cycle arrest and cellular apoptosis in cancer cells. The cell cycle arrest caused by ASTs was associated with increases in the expression levels of p21Cip1/Waf1, p27, and p53, as well as decreases in the levels of CDK4 and CDK6. Meanwhile, the apoptosis induced by ASTs was confirmed by activation of caspase-3 and PARP in the cancer cells. The results indicated that hydroxyl (OH) at C3 and C3' of terminal ring structure might not be the major factor that affects the anticancer activity of AST. This study revealed important information on the inhibitory effects of ASTs on human colon cancer cells, which provided a basis for using ASTs as chemopreventive agents for colon cancer.

MicroRNA-215 suppresses cell proliferation, migration and invasion of colon cancer by repressing Yin-Yang 1

Colorectal cancer is one of the most common malignant tumors worldwide with rising incidence. MicroRNAs are small non-coding RNAs that implicate in multiple physiological or pathological processes. The aberrant expression of miRNA-215 (miR-215) has been illustrated in various types of cancers. However, the expression of miR-215 in human colon cancer and the biological roles of it remain largely unknown. We conducted this study to explore the expression and the function of miR-215 in human colon cancer. The results showed that miR-215 was remarkably downregulated in colon cancer tissues and cell lines. Overexpression of miR-215 by miR-215 mimic significantly inhibited colon cancer cell proliferation, migration and invasion while knockdown of miR-215 by miR-215 inhibitor exerted reverse effects. Furthermore, we newly identified Yin-Yang 1(YY1) as a direct target of miR-215 which could rescue the effects of miR-215 on colon cancer cells. In summary, our investigation revealed that miR-215 was downregulated in colon cancer and it suppressed colon cancer cell proliferation, migration and invasion by directly targeting YY1.

Water-Based Chitosan for Thymine Conjugation: A Simple, Efficient, Effective, and Green Pathway to Introduce Cell Compatible Nucleic Acid Recognition.

Chitosan is a potential biopolymer for cell recognition and targeting; however, when those functions are based on cationic amine groups of chitosan, cell damage is a concern. This study presents water-based chitosan conjugated with thymine (CsT) through a mild and homogeneous conjugating reaction via amide bond without the use of organic and/or acidic solvents. The CsT displays water-solubility in a wide range of pH. A series of comparative gel retardation assays confirm the selective binding with poly(A), resulting in nanoparticles of 100 to 250 nm in size. PrestoBlue cell viability assay clarifies nontoxicity and reveals noncytotoxicity to normal colon cells but inhibition of colon cancer cells. This simple pathway for water-soluble chitosan-nucleic acid leads to synergistic effects of cell compatibility and DNA recognition.

γ-Aminobutyric acid inhibits the proliferation and increases oxaliplatin sensitivity in human colon cancer cells.

γ-Aminobutyric acid (GABA) is a natural non-protein amino acid, which broadly exists in many plant parts and is widely used as an ingredient in the food industry. In mammals, it is widely distributed in central nervous system and non-neural tissues. In addition to a primary inhibitory neurotransmitter in the central nervous system, endogenous GABA content has been found to be elevated in neoplastic tissues in colon cancer. However, the effect of extraneous GABA on colon cancer has rarely been reported. In this study, we found the inhibitory effects of GABA on the proliferation of colon cancer cells (CCCs). The amino acid also suppressed metastasis of SW480 and SW620 cells. To further study the correlated mechanism, we analyzed the changes in cell cycle distribution and found that GABA suppressed cell cycle progression through G2/M or G1/S phase. Furthermore, RNA sequencing analysis revealed GABA-induced changes in the mRNA expression of 30 genes, including EGR1, MAPK4, NR4A1, Fos, and FosB, in all the three types of CCC. Importantly, GABA enhanced the anti-tumor efficacy of oxaliplatin (OXA) in subcutaneous xenograft tumor model in nude mice. The data suggest that GABA inhibits colon cancer cell proliferation perhaps by attenuating EGR1-NR4A1 axis, EGR1-Fos axis, and by disrupting MEK-EGR1 signaling pathway. This work reveals the pharmacological value of GABA derived from food and suggests that exogenous GABA might play an auxiliary role in polychemotherapy of colon cancer.

Curcumin suppresses colon cancer cell invasion via AMPK-induced inhibition of NF-κB, uPA activator and MMP9.

Curcumin, an active nontoxic ingredient of turmeric, possesses potent anti-inflammatory, antioxidant and anti-cancer properties; however, the molecular mechanisms of curcumin are not fully understood. The transcription factor nuclear factor-κB (NF-κB) is key in cellular processes, and the expression/activation of urokinase-type plasminogen activator (uPA) and matrix metalloproteinase-9 (MMP9) are crucial for cell invasion. The present study investigated the hypothesis that curcumin inhibits colon cancer cell invasion by modulating NF-κB-mediated expression and activation of uPA and MMP9. Human colon cancer SW480 and LoVo cells were treated with various concentrations of curcumin. Curcumin was demonstrated to dose-dependently inhibit the adhesion and proliferation ability of LoVo and SW480 cells using Transwell and MTT assays, respectively. In addition, curcumin activated 5' AMP-activated protein kinase (AMPK) and suppressed p65 NF-κB phosphorylation, as shown by western blot analysis. Compound C, a potent AMPK inhibitor, abolished curcumin-induced inhibition of NF-κB, uPA and MMP9, suggesting that AMPK activation is responsible for curcumin-mediated NF-κB, uPA and MMP9 inhibition. The binding activity of NF-κB to DNA was examined and western blotting and quantitative polymerase reaction was performed to detect the effect of curcumin on the expression of uPA and MMP9. The present results revealed that curcumin significantly decreased the expression of uPA and MMP9 and NF-κB DNA binding activity. Furthermore, curcumin decreased the level of the p65 subunit of NF-κB binding to the promoter of the gene encoding uPA and MMP9, which suppressed transcriptional activation of uPA and MMP9. Overall, the present data suggest that curcumin inhibits colon cancer cell invasion via AMPK activation and subsequent inhibition of p65 NF-κB, uPA and MMP9. The therapeutic potential of curcumin for colon cancer metastasis required additional study.