SW480 Human Colon Cancer Cells Research Articles

Ergone Derivatives from the Deep-Sea-Derived Fungus Aspergillus terreus YPGA10 and 25,28-Dihydroxyergone-Induced Apoptosis in Human Colon Cancer SW620 Cells.

Ten new ergone derivatives (1-10) and five known analogues (11-15) were isolated from the deep-sea-derived fungus Aspergillus terreus YPGA10. The structures including the absolute configurations were established by detailed analysis of the NMR spectroscopic data, HRESIMS, ECD calculation, and coupling constant calculation. All the structures are characterized by a highly conjugated 25-hydroxyergosta-4,6,8(14),22-tetraen-3-one nucleus. Structurally, compound 2 bearing a 15-carbonyl group and compounds 5-7 possessing a 15β-OH/OCH3 group are rarely encountered in ergone derivatives. Bioassay results showed that compounds 1 and 11 demonstrated cytotoxic effects on human colon cancer SW620 cells with IC50 values of 8.4 and 3.1 μM, respectively. Notably, both compounds exhibited negligible cytotoxicity on the human normal lung epithelial cell BEAS-2B. Compound 11 was selected for preliminary mechanistic study and was found to inhibit cell proliferation and induce apoptosis in human colon cancer SW620 cells. In addition, compound 1 displayed cytotoxic activity against five human leukemia cell lines with IC50 values ranging from 5.7 to 8.9 μM. Our study demonstrated that compound 11 may serve as a potential candidate for the development of anticolorectal cancer agents.

Human β-defensin-1 affects the mammalian target of rapamycin pathway and autophagy in colon cancer cells through long non-coding RNA TCONS_00014506.

Colorectal cancer has a low 5-year survival rate and high mortality. Human β-defensin-1 (hBD-1) may play an integral function in the innate immune system, contributing to the recognition and destruction of cancer cells. Long non-coding RNAs (lncRNAs) are involved in the process of cell differentiation and growth. To investigate the effect of hBD-1 on the mammalian target of rapamycin (mTOR) pathway and autophagy in human colon cancer SW620 cells. CCK8 assay was utilized for the detection of cell proliferation and determination of the optimal drug concentration. Colony formation assay was employed to assess the effect of hBD-1 on SW620 cell proliferation. Bioinformatics was used to screen potentially biologically significant lncRNAs related to the mTOR pathway. Additionally, p-mTOR (Ser2448), Beclin1, and LC3II/I expression levels in SW620 cells were assessed through Western blot analysis. hBD-1 inhibited the proliferative ability of SW620 cells, as evidenced by the reduction in the colony formation capacity of SW620 cells upon exposure to hBD-1. hBD-1 decreased the expression of p-mTOR (Ser2448) protein and increased the expression of Beclin1 and LC3II/I protein. Furthermore, bioinformatics analysis identified seven lncRNAs (2 upregulated and 5 downregulated) related to the mTOR pathway. The lncRNA TCONS_00014506 was ultimately selected. Following the inhibition of the lncRNA TCONS_00014506, exposure to hBD-1 inhibited p-mTOR (Ser2448) and promoted Beclin1 and LC3II/I protein expression. hBD-1 inhibits the mTOR pathway and promotes autophagy by upregulating the expression of the lncRNA TCONS_00014506 in SW620 cells.

The non-vesicle cell-free DNA (cfDNA) induces cell transformation associated with horizontal DNA transfer.

Cell-free DNA (cfDNA) is a source for liquid biopsy used for cancer diagnosis, therapy selection, and disease monitoring due to its non-invasive nature and ease of extraction. However, cfDNA also participates in cancer development and progression by horizontal transfer. In humans, cfDNA circulates complexed with extracellular vesicles (EV) and macromolecular complexes such as nucleosomes, lipids, and serum proteins. The present study aimed to demonstrate whether cfDNA not associated with EV induces cell transformation and tumorigenesis. Supernatant of the SW480 human colon cancer cell line was processed by ultracentrifugation to obtain a soluble fraction (SF) and a fraction associated with EV (EVF). Primary murine embryonic fibroblast cells (NIH3T3) underwent passive transfection with these fractions, and cell proliferation, cell cycle, apoptosis, cell transformation, and tumorigenic assays were performed. Next, cfDNA was analyzed by electronic microscopy, and horizontal transfer was assessed by human mutantKRASin recipient cells via PCR and recipient cell internalization via fluorescence microscopy. The results showed that the SF but not the EVF of cfDNA induced proliferative and antiapoptotic effects, cell transformation, and tumorigenesis in nude mice, which were reduced by digestion with DNAse I and proteinase K. These effects were associated with horizontal DNA transfer and cfDNA internalization into recipient cells. The results suggest pro-tumorigenic effects of cfDNA in the SF that can be offset by enzyme treatment. Further exploration of the horizontal tumor progression phenomenon mediated by cfDNA is needed to determine whether its manipulation may play a role in cancer therapy.

Potential health benefits of combined extracts from germinated brown jasmine rice (Oryza sativa L.), Moringa oleifera leaves, and Cordyceps militaris

ABSTRACT Extracts from powders of geminated brown rice (Oryza sativa L.), Moringa oleifera, and Cordyceps militaris were combined in various ratios and their health-beneficial properties were investigated. All combined extracts exerted the synergistic effects on antioxidant activity as well as inhibitions against α-amylase and α-glucosidase. Extract formulas T4 (70% germinated brown jasmine rice, 20% M. oleifera dry leaves, and 10% C. militaris) and T2 (80% germinated brown jasmine rice, 10% M. oleifera dry leaves, and 10% C. militaris) elicited the significantly highest antioxidant activity and the significantly highest inhibition against α-glucosidase, respectively. Both formulas exhibited the significantly highest inhibition against α-amylase. As compared to each other, extract formulas T4 and T2 possessed the higher glucose consumption-promoting and antiinflammatory activities, respectively. Both formulas were not toxic to normal RAW-264.7 and SW-480 human colon cancer cells. Bioactive compounds with health-supporting properties were identified using UHPLC-QTOF-MS/MS. These combined extracts can be promising therapeutic food.

Luteolin exhibits synergistic therapeutic efficacy with erastin to induce ferroptosis in colon cancer cells through the HIC1-mediated inhibition of GPX4 expression

Colon cancer continues to be a prevalent gastrointestinal malignancy with a bleak prognosis. The induction of ferroptosis, a new form of regulated cell death, has emerged as a potentially effective strategy for the treatment of colon cancer. However, numerous colon cancer cells display resistance to ferroptosis induced by erastin, a well-established ferroptosis inducer. Finding drugs that can enhance the susceptibility of colon cancer cells to erastin is of utmost importance. This study aimed to examine the synergistic therapeutic impact of combining erastin with a bioactive flavonoid compound luteolin on the ferroptosis-mediated suppression of colon cancer. Human colon cancer HCT116 and SW480 cells were used for the in vitro studies and a xenograft of colon cancer model in BALB/c nude mice was established for the in vivo experiments. The results showed that combinative treatment of luteolin and erastin effectively inhibited the viability and proliferation of colon cancer cells. Luteolin and erastin cotreatment synergistically induced ferroptosis, concomitant with a reduction in glutathione and an elevation in lipid peroxides. In vivo, combinative treatment of luteolin and erastin exhibited a pronounced therapeutic effect on xenografts of colon cancer, characterized by a significant induction of ferroptosis. Mechanistically, luteolin in combination with erastin synergistically reduced the expression of glutathione peroxidase 4 (GPX4), an antioxidase overexpressed in colon cancer cells. Furthermore, luteolin and erastin cotreatment significantly upregulated the expression of hypermethylated in cancer 1 gene (HIC1), a transcriptional repressor also recognized as a tumor suppressor. HIC1 overexpression notably augmented the suppression of GPX4 expression and facilitated ferroptotic cell death. In contrast, HIC1 silencing attenuated the inhibition of GPX4 expression and eliminated the ferroptosis. Conclusively, these results clearly demonstrated that luteolin acts synergistically with erastin and renders colon cancer cells vulnerable to ferroptosis through the HIC1-mediated inhibition of GPX4 expression, which may act as a promising therapeutic strategy.

Design, synthesis, and anticancer activity of novel 4,6-dimorpholinyl-1,3,5-triazine compounds

Abstract A series of novel 4,6-dimorpholinyl-1,3,5-triazine derivatives 6a–6r were obtained through N-substitution and Claisen-Schmidt condensation. 1H NMR, 13C NMR, and mass spectrometry were used to characterize the molecular structures of the derivatives. The in vitro antiproliferation activity of derivatives was evaluated using the MTT assay against SW620 (human colon cancer cells), A549 (human nonsmall cell lung cancer cells), HeLa (human cervical cancer cells), and MCF-7 (human breast cancer cells). Compound 6o bearing a pyridyl group exhibited good cytotoxicity against four cancer cells, with IC50 values of 8.71, 9.55, 15.67, and 21.77 μM, sequentially. In addition, compound 6a showed some selectivity against SW620.

Hypoxia Increases ATX Expression by Histone Crotonylation in a HIF-2α-Dependent Manner.

Autotaxin (ATX), the key enzyme that generates lysophosphatidic acid (LPA) from lysophosphatidylcholine (LPC), is involved in tumorigenesis through the ATX-LPA axis and is regarded as a valuable target in tumor therapy. Hypoxia is a major feature of solid tumors and contributes to tumor development with striking alterations in the gene expression profile. Here, we show that hypoxia induces ATX expression in a hypoxia-inducible factor (HIF) 2α-dependent fashion in human colon cancer SW480 cells. HIF-2α is directly bound to specific hypoxia response elements (HREs) in the ATX promoter. Under hypoxic conditions, knockout or inhibition of ATX suppressed the migration of SW480 cells, which could be rescued by the addition of LPA, suggesting that the induction of ATX during hypoxia promotes cancer cell migration through the ATX-LPA axis. Further studies showed that ATX expression was induced by HIF-2α through recruiting p300/CBP, which led to crotonylation but not acetylation of histone H3 in the ATX promoter region during hypoxia. Moreover, elevation of cellular histone crotonylation levels could induce ATX expression under normoxic conditions. In conclusion, our findings reveal that ATX is induced in SW480 cells during hypoxia by histone crotonylation in a HIF-2α-dependent manner, while as a novel mechanism of ATX expression regulation, the upregulation of ATX expression by histone crotonylation is not confined to hypoxia.

Chinese herb formulae inhibit the proliferation of human colon cancer SW480 cells by inducing cell apoptosis

Objective: This study aimed to reveal the antitumor effects of Chinese herbal formulae and the underlying mechanisms in treating colorectal cancer, with a focus on developing traditional Chinese medicine (TCM) as a supplement and alternative therapeutic method for cancers. Materials and Methods: Human colon cancer SW480 cells were treated with three Chinese herbal formulae, Bu Zhong Yi Qi Decoction, Fuzi Lizhong Decoction, and Pulsatilla Decoction at different concentrations (50–600 μg/mL) for 24, 36, and 48 h, respectively. Cell viability was determined using the resazurin reduction assay, and cell survival rate was evaluated using a colony formation assay. After treatment with different concentrations (50–600 μg/mL) of these three formulae for 48 h, the effects of the Chinese herbal formulae on cell apoptosis were investigated using Hoechst/propidium iodide (PI) staining. The positive PI-stained cells were investigated using an EnSpire multilabel plate reader and the positive Hoechst-stained cells were observed under a fluorescence microscope for morphological changes. Results: Bu Zhong Yi Qi Decoction, Fuzi Lizhong Decoction, and Pulsatilla Decoction inhibited SW480 cell proliferation in a dose-and time-dependent manner and induced cell apoptosis. Conclusion: Chinese herbal formulae with a special prescription form of TCM with antitumor effects bring a new perspective in line with the principles of TCM in cancer treatment.

Gene Expression of Effect of Combination Cassia Fistula Extract and Cisplatin on Inhibition of Human Colon Cancer SW480 Cell Line

Gene Expression of Effect of Combination Cassia Fistula Extract and Cisplatin on Inhibition of Human Colon Cancer SW480 Cell Line

Copper Nanoparticles Induce Apoptosis and Oxidative Stress in SW480 Human Colon Cancer Cell Line.

Cu nanoparticles (CuNPs) have various applications in biomedicine, owing to their unique properties. As the effect of CuNPs on the induction of oxidative stress and apoptosis in the human colorectal cancer cell line SW480 has not yet been studied, we investigated the toxicity and mechanism of action of these NPs in SW480 cells. MTT assay was performed to assess the effect of the particles on the viability of SW480 cells. The levels of oxidative stress were assessed after 24h of treatment with CuNPs by evaluating the Reactive Oxygen Specious (ROS) production. The antioxidant enzyme activity was assessed using a colorimetric method. To investigate the effect of NPs on cellular apoptosis, Hoechst33258 staining was performed, and the expression of Bax, Bcl-2, and p53 was evaluated by qRT-PCR. The MTT assay results showed that CuNPs inhibited the viability of SW480 cells. Moreover, the increase in ROS production at all three concentrations (31, 68, and 100μg/ml) was significant. It has been observed that CuNPs lead to increased expression of Bax and p53, and decreased expression of Bcl-2. Hoechst staining was performed to confirm apoptosis. In conclusion, the induction of apoptosis demonstrated the anticancer potential of the CuNPs.

Design, synthesis and antitumor activity evaluation of novel indole acrylamide derivatives as IMPDH inhibitors

Inosine 5′-monophosphate dehydrogenase (IMPDH; EC1.1.1.205) is the rate-limiting enzyme of GMP biosynthesis. The inhibition of IMPDH limits the growth and survival of tumors. Based on the systematic summary of clinical IMPDH inhibitors, 38 acrylamide derivatives with differently substituted indoles at the 3-position as the core scaffold were designed and synthesized. In addition, the actions of these compounds at the enzyme and cellular levels were evaluated. An MTT assay with different kinds of cells was used to assess the cytotoxic activities of compounds 14e and 14n, which displayed potent hIMPDH2 inhibitory activities (IC50 = 4.207 and 2.948 μM, respectively). Biological evaluation indicated that target compounds 14e and 14n displayed the most significant effects on SW480 human colon cancer cells (IC50 = 15.34 ± 0.06 and 15.31 ± 0.09 μM, respectively), and it was determined that these compounds are effective and valuable IMPDH inhibitors for cancer intervention.

Cucurbitane-type triterpenoids from the branches and leaves of Elaeocarpus sylvestris

The chemical structures of nine cucurbitane-type triterpenoids from the branches and leaves of Elaeocarpus sylvestris (Elaeocarpaceae), including undescribed 29-hydroxymogroside I E 2 , epimogroside I E 2 , epimogroside I E 1 , 24-oxomogroside I E 1 , and 11- O -acetylmogroside I E 1 , were determined using mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy. The absolute configuration of 29-hydroxymogroside I E 2 was confirmed by performing an X-ray diffraction analysis. Cucurbitacin D and 11- O -acetylmogroside I E 1 showed cytotoxicity toward human leukemia HL-60, human lung adenocarcinoma A549, human hepatoma SMMC-7721, human breast cancer MCF-7, and human colon cancer SW480 cell lines. The presence of a 11- O -acetyl group might increase the cytotoxicity of this type of triterpenoids. • Five undescribed cucurbitane-type triterpenoids were isolated from Elaeocarpus sylvestris. • Cucurbitacin D and 11-O-acetylmogroside I E 1 showed cytotoxicity toward five cancer cell lines. • The presence of a 11-O-acetyl group might increase the cytotoxicity of this type of triterpenoids.

Gnetum montanum extract induces apoptosis by inhibiting the activation of AKT in SW480 human colon cancer cells

Context Gnetum montanum Markgr. (Gnetaceae) is used to treat rheumatic arthralgia and bruises in the clinic. Objective To exam the activity and mechanism of G. montanum extract (GME) against colon cancer cells SW480. Materials and methods The anti-proliferative activity of GME (0–120 μg/mL) on SW480 cells was determined using MTS assay at 24, 48, and 72 h. The in vitro activity of GME (0–120 μg/mL) on SW480 cells was investigated using flow cytometry and western blotting analysis. The in vivo activity of GME was evaluated using xenograft tumour model of zebrafish and nude mice. The chemical composition of GME was detected by using HPLC–MS/MS. Results The IC50 value SW480 cells viability by GME were 126.50, 78.25, and 50.77 μg/mL, respectively, for 24, 48, and 72 h. The experiments showed that apoptotic cells and G2/M phase cells increased from 20.81 to 61.53% (p < 0.01) and 25.76 to 34.93% with 120 μg/mL GME, respectively. GME also down-regulated the protein expression of P-AKT, P-GSK-3β, P-PDK1, P-c-Raf, caspase-3, and Bcl-2, and up-regulated the expression cleaved caspase-3, cleaved PARP, and Bax. In vivo study found that GME can significantly inhibit the growth and migration of SW480 cells in xenograft zebrafish. GME reduced the nude mice tumour weight to approximately 32.19% at 28 mg/kg/day and to 53.17% (p < 0.01) at 56 mg/kg/day. Forty-two compounds were identified from the GME. Discussion and conclusions GME has a significant antitumor effect on colon cancer cells SW480, and it has the potential to be developed as an anticancer agent.

The Impact of Sorghum Polyphenols on DNA Methylation and Signaling Pathways in Colon Cancer

Colorectal cancer (CRC) is a major public health issue, accounting for nearly 150,000 cases annually. Because CRC correlates with inflammation and oxidative stress, exploiting the anti‐oxidative and anti‐inflammatory nature of bioactive components in our diet, such as polyphenols, could potentially contribute to the inhibition of cancer initiation. Polyphenols are an immense family of naturally occurring secondary plant metabolites found in fruits, vegetables, and cereal grains, and have been demonstrated across many in vitro and in vivo studies to protect against carcinogenesis. Not only are polyphenols implicated to impact cancer development but they may also be involved in epigenetic regulation. Abnormalities in epigenetic modifications can lead to aberrant regulation of gene expression, and depending on the genes in question, this may contribute to the cell’s ability to acquire hallmarks of cancer. New varietals of the grain Sorghum bicolor provided by the USDA contain high amounts of polyphenols, which may contribute to CRC prevention due to their strong antioxidative and anti‐carcinogenic properties. The expression of genes involved in the promotion or prevention of CRC, like the Wnt/β‐catenin signaling pathway, may be regulated via epigenetic mechanisms. We hypothesize that the high polyphenol sorghum extracts may decrease DNA methylation, thus impacting the expression genes involved in the Wnt/β‐catenin signaling pathway. For our current study, we treated HCT116 and SW480 human colon cancer cells with novel high‐polyphenol S. bicolor extracts (HP and SC) or an extract of low‐polyphenol Macia‐Sorghum for up to 48 hours. Controls included cells treated with solvent or 5 µM epigallocatechin gallate as a benchmark. Total RNA was harvested, and the mRNA expression of selected genes was quantitated via qPCR. Relative to solvent control, the high‐polyphenol S. bicolorextract HP increased mRNA expression of DNA methyltransferase 1 (DNMT1), the key enzyme that upholds methylation patterns after DNA synthesis. Furthermore, mRNA expression of DNMT3A, the enzyme responsible for de novo DNA methylation, was also increased. The SC extract significantly decreased the expression of β‐catenin while increasing expression of transcription factor 4 (TCF4), a gene that mediates the expression of pro‐tumorigenic Wnt target genes such as VEGF and cMYC. Encouraged by these preliminary findings, we will investigate mRNA and protein expression of genes that are involved in epigenetic mechanisms, as well as those of downstream signaling pathways. Our study may help elucidate the question, whether consumption of high‐polyphenol S. bicolor as part of the human diet may lead to beneficial changes in DNA methylation. Subsequently, the expression of specific genes involved in the promotion or prevention of colorectal cancer may be modified, providing a dietary intervention to CRC.

Cytotoxicity of green-synthesized silver nanoparticles by Adansonia digitata fruit extract against HTC116 and SW480 human colon cancer cell lines

Abstract Nanoparticles synthesized from plants are being explored for cancer treatment therapies all over the world. This study reported the eco-friendly and low-cost method for the green synthesis of silver nanoparticles (AgNPs) from Adansonia digitata fruit as a reducing and capping agent. The anti-cancer potential of synthesized particles was explored against HTC116 and SW480 colon cancer cell lines. Prepared AgNPs were characterized by ultraviolet-visible spectroscopy, zeta potential, transmission electronic microscopy, scanning electronic microscopy, Fourier transform infrared, and energy-dispersive spectrum. The cytotoxicity was determined with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and expression levels of four genes (CTNNB1, APC, LRP5, and LRP6) were checked by reverse transcription polymerase chain reaction. The sharp peak of surface plasmon resonance at 400 nm confirms the formation of AgNPs. Dynamic light scattering showed average sizes of 16.34 nm with a polydispersity index of 0.193. A. digitata AgNPs were spherical with slight aggregated. AgNPs were more cytotoxic than A. digitata extract and decrease the expression of CTNNB1 and LRP6 genes while LRP5 gene expression was increased in both cell lines. APC gene expression was decreased in SW480 but increased in HTC116 with treatment. Overall, this study suggested that AgNPs synthesized by A. digitata fruit extract can be an attractive candidate for anticancer applications.

Brazilin From Caesalpinia sappan L. Induced Apoptosis via mTOR and HO-1 Pathway in SW480 Human Colon Cancer Cells

The mTOR pathway is a crucial biological regulatory mechanism of cell growth, proliferation and cell death, and its inhibitors were new candidates of anticancer drugs through regulation of energy balance and metabolism. In the present study, whether brazilin and mTOR inhibitor (Torin1) exerts anti-cancer effects was evaluated and the mechanism of its regulation in colorectal cancer cells investigated. Brazilin showed dose- and time-dependent cytotoxicity of colorectal cancer cells (SW480 cells) through apoptosis pathways such as Bcl-2, Bax, as well as cleavage of caspase 3, caspase 9, and PARP1. In addition, brazilin reduced mammalian target of rapamycin (mTOR) phosphorylation in a dose- and time-dependent manner, and the mTOR inhibitor torin 1 blocked this phosphorylation. Brazilin also decreased heme oxygenase-1 (HO-1) expression in a dose- and time-dependent manner; however, hemin, a specific HO-1 substrate, markedly increased HO-1 expression. Torin 1 reduced hemin-induced HO-1 expression and increased colorectal cell death in a dose-dependent manner in the presence and absence of hemin. Moreover, nuclear factor erythroid 2–related factor 2 (Nrf2) translocation into nucleus fraction was crucial role in brazilin-mediated apoptosis of colorectal cancer cells. These results showed that brazilin and torin1 might regulate the mTOR signaling pathway by decreasing mTOR phosphorylation. Furthermore, mTOR signaling was associated with brazilin-regulated HO-1 expression, which induced apoptosis in colorectal cancer cells. These results suggest that synthetic and/or natural mTOR inhibitors were useful candidate for treatment of colorectal cancer cells.

Effects of miR-490-5p targeting CDK1 on proliferation and apoptosis of colon cancer cells via ERK signaling pathway.

The aim of this study was to investigate the effects of micro ribonucleic acid (miR)-490-5p on the proliferation and apoptosis of colon cancer cells, and to explore its potential mechanism. The mRNA expression of miR-490-5p in 30 pairs of colon cancer tissues and adjacent normal tissues was detected via reverse transcription-polymerase chain reaction (RT-PCR). Human colon cancer SW480 cell lines were cultured in vitro and divided into Control group and miR-490-5p overexpression group (miR-490-5p mimic group). The nonsense sequence and miR-490-5p mimic were transfected using liposome transfection technique into colon cancer cells in control group and miR-490-5p mimic group, respectively. Cell proliferation and apoptosis in each group were then observed. At the same time, the effect of miR-490-5p on the growth of colon cancer in vivo was explored using subcutaneous tumorigenesis assay. The protein expressions of extracellular signal-regulated kinase (ERK)1/2 signaling pathway and cyclin-dependent kinase 1 (CDK1) were determined via Western blotting. Furthermore, immunohistochemical staining was performed to verify the protein expression of CDK1 in vivo. The expression of miR-490-5p in colon cancer tissues was significantly lower than that in adjacent normal tissues (p<0.05). After transfection with miR-490-5p mimic in vitro, EdU staining and colony formation assay showed that the proliferation ability of SW480 cells was significantly weakened (p<0.05). Meanwhile, the number of colonies in miR-490-5p mimic group was markedly less than that in Control group (p<0.05). The results of Western blotting revealed that overexpression of miR-490-5p remarkably up-regulated the Bax/Bcl-2 and C-Caspase3/T-Caspase3 ratios in cancer cells (p<0.05). Subsequent results indicated that the subcutaneous tumorigenesis of colon cancer cells was markedly inhibited by overexpression of miR-490-5p (p<0.05). According to the results of Western blotting, the activation of ERK signaling pathway and the protein expression of CDK1 were significantly suppressed by overexpression of miR-490-5p (p<0.05). In vivo experiments further revealed that the protein expression of CDK1 in colon cancer tissues increased significantly (p<0.05). MiR-490-5p was found lowly expressed in colon cancer patients. In addition, overexpression of miR-490-5p inhibited the proliferation and promoted the apoptosis of colon cancer cells via down-regulating CDK1 both in vitro and in vivo.

Aurora kinase A induces migration and invasion by inducing epithelial-to-mesenchymal transition in colon cancer cells

Aurora kinase is a family of serine/threonine kinases intimately associated with mitotic progression and the development of human cancers. Studies have shown that aurora kinases are important for the protein kinase C (PKC)-induced invasion of colon cancer cells. Recent studies have shown that aurora kinase A promotes distant metastasis by inducing epithelial-to-mesenchymal transition (EMT) in colon cancer cells. However, the role of aurora kinase A in colon cancer metastasis remains unclear. In this study, we investigated the effects of aurora kinase A on PKC-induced cell invasion, migration, and EMT in human SW480 colon cancer cells. Treatment with 12-O-tetradecanoylphorbol-13-acetate (TPA) changed the expression levels of EMT markers, increasing α-SMA, vimentin, and MMP-9 expression and decreasing E-cadherin expression, with changes in cell morphology. TPA treatment induced EMT in a PKC-dependent manner. Moreover, the inhibition of aurora kinase A by siRNAs and inhibitors (reversine and VX-680) suppressed TPA-induced cell invasion, migration, and EMT in SW480 human colon cells. Inhibition of aurora kinase A blocked TPA-induced vimentin and MMP-9 expression, and decreased E-cadherin expression. Furthermore, the knockdown of aurora kinase A decreased the transcriptional activity of NF-κB and AP-1 in PKC-stimulated SW480 cells. These findings indicate that aurora kinase A induces migration and invasion by inducing EMT in SW480 colon cancer cells. To the best of our knowledge, this is the first study that showed aurora kinase A is a key molecule in PKC-induced metastasis in colon cancer cells.

Component Analysis and Anti-Colorectal Cancer Mechanism via AKT/mTOR Signalling Pathway of Sanghuangporus vaninii Extracts.

Sanghuangporus vaninii (Ljub.) L.W. Zhou & Y.C. Dai (SV) is a major cultivar of Sanghuang, which is well known as an excellent anti-tumour drug and reaches the mainstream market in China. Water, 60% ethanol and 95% ethanol were used to extract the drug, and three kinds of polar extracts were obtained separately. Compared with water extracts and 95% ethanol extracts, the 60% ethanol extract had the highest flavonoid content, and its polysaccharide content was greater than that in the 95% ethanol extract and lower than that in the water extract. Its essential components were phenolics whose majority were phenolic acids, flavonoids and phenylpropanoids. This extract has better inhibition effects on the proliferation of SW480 human colon cancer cells, inducing cell apoptosis and blocking G2/M period cells. It can significantly inhibit gene expression and reduce the activation of the AKT/mTOR signalling pathway. The anti-cancer activity of the 60% ethanol extract is satisfactory and may be a result of the combined effects of polysaccharides and flavonoids. The data suggest that the 60% ethanol extract can be used as an adjuvant for chemotherapy and as a potential anti-cancer agent with broad development prospects.

Inhibitory Effect of Polyphyllin D on Proliferation of Human Colon Cancer Cell Line SW480 via the MicroRNA-21/Phosphatase and Tensin Homolog Deleted on Chromosome 10

To evaluate the effects of polyphyllin D on the proliferation and apoptosis of human colon cancer SW480 cell lines and to reveal whether this process is mediated by the micro ribonucleic acid-21/phosphatase and tensin homolog deleted on chromosome 10 pathway. Human colon cancer SW480 cells were transfected with micro ribonucleic acid-21-mimic or mimic-negative control and treated with polyphyllin D at different concentrations (control, 0.25, 0.5, 1 and 2 μM) for 24 h or 48 h. Compared with control polyphyllin D group, the viability of human colon cancer SW480 cells significantly declined but the apoptosis rate significantly rose in 0.5 μM polyphyllin D group and the colony number of SW480 cells significantly decreased in 0.25 μM polyphyllin D group (p<0.05). Compared with control group, 2 μM polyphyllin D group had significantly increased protein expressions of Bcl-2 associated X protein and cleaved caspase-3, but decreased protein expression of B-cell lymphoma-2 in SW480 cells (p<0.001). The protein levels of micro ribonucleic acid-21 and phosphorylated-protein kinase B in SW480 cells were significantly lower, while the phosphatase and tensin homolog deleted on chromosome 10 messenger ribonucleic acid and protein levels were higher in 2 μM polyphyllin D group than those in control group (p<0.001). However, the protein expression of protein kinase B was not changed in 2 μM polyphyllin D group (p>0.05). After transfection with micro ribonucleic acid-21-mimic, the level of micro ribonucleic acid-21 in SW480 cells was significantly higher, while the phosphatase and tensin homolog deleted on chromosome 10 messenger ribonucleic acid and protein levels were significantly lower than those in control group (p<0.001). Compared with control group, the antiproliferative and pro-apoptotic effects of polyphyllin D on SW480 cells were reversed by transfection with micro ribonucleic acid-2-mimic (p<0.001). Polyphyllin D inhibits proliferation and induces apoptosis of human colon cancer SW480 cells via the micro ribonucleic acid-21/phosphatase and tensin homolog deleted on chromosome 10 pathway.