Proliferation Of HCT116 Cells Research Articles

MicroRNA-142-3p Inhibits Tumorigenesis of Colorectal Cancer via Suppressing the Activation of Wnt Signaling by Directly Targeting to β-Catenin.

BackgroundAltered expression profile of microRNAs (miRNAs) was reported to be associated with colorectal cancer (CRC). The aims of this study are to identify the changed miRNAs in the plasma of CRC patients and explore the underlying mechanism of these miRNAs during tumorigenesis.MethodsPlasma miRNA expression profiles were compared between healthy people and CRC patients. MiRNA expression was measured using quantitative real-time PCR. Colony formation and MTT assays were used to test cell proliferation. Luciferase assay, immunohistochemistry and Western blotting were employed to explore the molecular mechanism.ResultsMiR-142-3p level was found as the most significantly repressed miRNA in CRC patients. Overexpression of miR-142-3p dramatically repressed colony formation and cell proliferation of both HT29 and HCT116 cells while inhibition of miR-142-3p promoted those of the cells. Interestingly, overexpression of miR-142-3p reduced the level and nuclear accumulation of β-catenin. We further observed that miR-142-3p remarkably inhibited the transcriptional activity of β-catenin gene (CTNNB1). However, mutations in the predicted binding sites blocked this inhibition, suggesting that miR-142-3p may directly bind to the mRNA of β-catenin.ConclusionIn conclusion, we identified miR-142-3p exerts its function as a tumor suppressor through blocking the activation of Wnt signaling by directly targeting to CTNNB1.

Multi-spectroscopic and molecular modeling studies on the interactions of serum albumin with 20(S, R)-protopanaxadiol and 20(S, R)-protopanaxatriol that inhibit HCT-116 cells proliferation

Serum albumin (SA) acts as the carrier protein to transport drugs and plays a crucial role in the pharmacokinetics of distribution. In this work, the cytotoxicities of 20(S, R)-protopanaxadiol [20(S, R)-PPD] and 20(S, R)-protopanaxatriol [20(S, R)-PPT] against HCT-116 cell lines were assessed. 20(S)-PPD, 20(R)-PPD, 20(S)-PPT, and 20(R)-PPT inhibited the cells growth in a concentration-dependent manner with the IC50 values of 30.27, 18.80, 68.28, and 14.38 μM, respectively. The result of ABTS assay showed that 20(S, R)-PPD-BSA exhibited a higher antioxidant activity than that of 20(S, R)-PPD alone. Subsequently, the binding affinity and interaction mechanism of ginsenosides with SA were studied by the ultraviolet–visible absorption and fluorescence spectroscopy, which indicated that ginsenosides interfered with the microenvironment around amide bonds in SA. Through the calculation of Stern-Volmer plot, the quenching rate kq values of 20(S)-PPD, 20(R)-PPD, 20(S)-PPT, and 20(R)-PPT were 6.75, 4.30, 2.37, and 3.31 ( × 1011 M−1 s−1), respectively. These results suggested that the quenching was not caused by dynamic collision, but the formation of ginsenoside-SA complexes. The results of molecular docking suggested that these compounds tended to bind more strongly to Sudlow's site II of HSA rather than Sudlow's site I. In addition, 20(S)-ginsenosides showed more stable binding to HSA in comparison with 20(R)-ginsenosides, which may help elucidating the mechanisms that account for the stronger bioactivities of 20(S)-ginsenosides.

Expression of NEK2 in Colorectal Cancer Tissue and Its Effect on Proliferation, Invasion and Migration of HCT116 Cells

Expression of NEK2 in Colorectal Cancer Tissue and Its Effect on Proliferation, Invasion and Migration of HCT116 Cells

Knockdown of AURKA sensitizes the efficacy of radiation in human colorectal cancer

AimsAbnormally amplified expression of AURKA (aurora kinase A) is closely related to chemo-resistance in human colorectal cancer, lung cancer and leukemia. However, the biological role of AURKA in response to radio-sensitivity in human colorectal cancer is still unknown. Therefore, we evaluated the radio-sensitize ability of perturbation AURKA in human colorectal cancer. Main methodsThe knockdown effect of shAURKA was determined by western blot and qRT-PCR, respectively. Cell growth was determined by CCK-8 and clonogenic assay. Cell migration and metastasis was measured by wound healing assay and transwell invasive assay, respectively. Cell cycle and apoptosis was analyzed by flow cytometry. The alteration of down-stream targets was determined by western blot analysis. Key findingsWe observed that high-level of AURKA expression is associated with poor prognosis in CRC patients receiving radiotherapy. Knockdown of AURKA significantly sensitizes the efficacy of radiation on the proliferation of HCT116 and HT-29 cells. The combination of AURKA inhibition and radiation could effectively suppress the ability of cell migration and metastasis, but also synergistically induce cellular apoptosis and arrest cell cycle at G2/M phase. Further studies demonstrated that knockdown AURKA markedly enhanced the efficacy of radiation through elevated PARP cleavage and induced AURKA-mediated pro-apoptosis factor BIM. Meanwhile, knockdown of AURKA in combination with radiation synergistically suppressed the regulator in blockage of G2/M phase, CDK2. SignificanceTaken together, our results provide the evidence that targeted inhibition of AURKA could be a promising strategy for enhancing the efficacy of radiation for the treatment of human colorectal cancer.

A short deletion in the DNA-binding domain of STAT3 suppresses growth and progression of colon cancer cells.

In this study, we investigated the effect of a short deletion in the DNA-binding domain of STAT3 (STAT3del) on the transcriptional activation of STAT3 target genes and its relationship with colon carcinogenesis. We used the CRISPR-CAS9 gene editing system to delete a short sequence encoding amino acids 400-411 in the DNA-binding domain (amino acid sequence: 317-567) from STAT3 gene in SW480, SW620 and HCT116 colon cancer cells. ChIP sequencing analysis showed that STAT3del occupancy was significantly reduced in 1029 genes and significantly increased in 475 genes compared to wild-type STAT3. The mutation altered the DNA motifs recognized by STAT3del as compared to the wild-type STAT3. We observed a strong correlation between expression of the STAT3 target genes and the loss or gain of STAT3del binding to their promoters. CCK-8, wound healing, and TUNEL assays showed reduced proliferation, migration, and survival of SW480, SW620 and HCT-116 cells expressing STAT3del as compared to the corresponding controls. These findings demonstrate that a short deletion in the DNA-binding domain of STAT3 alters its genome-wide DNA-binding and transcriptional profile of STAT3-target proteins, and suppresses the growth, progression and survival of colon cancer cells.

Long non-coding RNA OTUD6B-AS1 overexpression inhibits the proliferation, invasion and migration of colorectal cancer cells via downregulation of microRNA-3171.

Colorectal cancer (CRC) is a common digestive system malignancy and a major cause of cancer-associated mortality worldwide. Aberrant expression of long non-coding RNAs has been reported in several types of cancer. The aim of the present study was to investigate the role of ovarian tumor domain containing 6B antisense RNA1 (OTUD6B-AS1) in CRC and its underlying mechanisms. OTUD6B-AS1 expression in CRC cell lines was examined using reverse transcription-quantitative PCR. Furthermore, The Cancer Genome Atlas database was utilized to examine the expression levels of OTUD6B-AS1 in CRC tissues. Following OTUD6B-AS1 overexpression, Cell Counting Kit-8 and colony formation assays were used to detect the proliferation ability of HCT116 cells. The expression levels of proliferation-related protein Ki67 were determined using immunofluorescence staining. Subsequently, Transwell and wound healing assays were used to evaluate the invasion and migration of HCT116 cells, respectively. The expression levels of migration-related proteins (MMP2 and MMP9) were measured using western blotting. Additionally, a luciferase reporter assay was used to verify the potential interaction between OTUD6B-AS1 and microRNA-3171 (miR-3171). Subsequently, rescue assays were performed to clarify the regulatory effects of OTUD6B-AS1 and miR-3171 on CRC development. The results demonstrated that OTUD6B-AS1 expression was low in CRC cells and tissues. Overexpression of OTUD6B-AS1 inhibited the proliferation, invasion and migration of HCT116 cells. Furthermore, miR-3171 was demonstrated to be a direct target of OTUD6B-AS1 using a luciferase reporter assay. The rescue assays revealed that miR-3171 mimics markedly reversed the inhibitory effects of OTUD6B-AS1 overexpression on proliferation, invasion and migration of CRC cells. Overall, these findings demonstrated that OTUD6B-AS1 overexpression inhibited the proliferation, invasion and migration of HCT116 cells via downregulation of miR-3171, suggesting that OTUD6B-AS1 may serve as a novel biomarker for CRC treatment.

Overexpression of microRNA-29b inhibits epithelial-mesenchymal transition and angiogenesis of colorectal cancer through the ETV4/ERK/EGFR axis

BackgroundRecent studies have reported the involvement of microRNA-29 (miR-29) family members in human cancers through their ability to regulate cellular functions. The present study investigated biological function of miR-29b in colorectal cancer (CRC).MethodsCRC tissues and adjacent normal tissues were collected and the expression of ETV4 and miR-29b in the tissues were identified. The relationship between ETV4 and miR-29b or ETV4 expression and the EGFR promoter was identified using dual-luciferase reporter gene and CHIP assays. The proliferation, invasion, migration, and apoptosis of CRC HCT116 cells were assayed using MTT assay, Scratch test, Transwell assay, and flow cytometry, respectively. Also, expression of epithelial-mesenchymal transition (EMT) markers, angiogenic factors, and vasculogenic mimicry formation were evaluated using RT-qPCR and Western blot.ResultsETV4 was upregulated, while miR-29b expression was decreased in CRC tissues. ETV4 was identified as a target gene of miR-29b, which in turn inactivated the ERK signaling pathway by targeting ETV4 and inhibiting EGFR transcription. Transfection with miR-29b mimic, siRNA-ETV4, or ERK signaling pathway inhibitor U0126 increased expression of E-cadherin and TSP-1, and CRC cell apoptosis, yet reduced expression of ERK1/2, MMP-2, MMP-9, Vimentin, and VEGF, as well as inhibiting EMT, angiogenesis, and CRC cell migration and invasion. The EMT, angiogenesis and cancer progression induced by miR-29b inhibitor were reversed by siRNA-mediated ETV4 silencing.ConclusionsmiR-29b suppresses angiogenesis and EMT in CRC via the ETV4/ERK/EGFR axis.

Inhibition of Growth of Colon Tumors and Proliferation of HT-29 Cells by Warburgia ugandensis Extract through Mediating G0/G1 Cell Cycle Arrest, Cell Apoptosis, and Intracellular ROS Generation.

Warburgia ugandensis Sprague (W. ugandensis), widely distributed in Africa, is a traditional medicinal plant used for the treatment of various diseases including cancer. We intended to evaluate the anticolorectal cancer (CRC) activities of the crude extract from W. ugandensis (WUD) and reveal the underlying molecular mechanisms of its action. We found that WUD inhibited the proliferation of HT-29 and HCT116 cells in a time- and dose-dependent manner and induced intracellular ROS generation. The inhibitory effect of WUD on the proliferation of HT-29 and HCT116 cells could be attenuated by NAC (a ROS scavenger) in a dose-dependent manner. WUD induced G0/G1 phase arrest, down-regulated the protein expression of Cyclin D1 via ROS accumulation in HT-29 cells. In search of the molecular mechanism involved in WUD-induced Cyclin D1 down-regulation, it was found that WUD can suppress PI3K/Akt/GSK3β signaling pathway in HT-29 cells. Next, it was found that WUD also activated apoptosis, poly-ADP ribose polymerase 1 (PARP1) cleavage and down-regulated pro-caspase 3 in HT-29 and HCT116 cells. Besides, WUD decreased the growth of colon tumors in vivo in the xenograft mouse model. We demonstrated for the first time that ROS and their modulation in the corresponding intracellular signaling could play a significant role in the potential activity of WUD against CRC cells.

The LncRNA FEZF1-AS1 promotes tumor proliferation in colon cancer by regulating the mitochondrial protein PCK2.

LncRNAs and metabolism represents two factors involved in cancer initiation and progression. However, the interaction between lncRNAs and metabolism remains to be fully explored. In this study, lncRNA FEZF1-AS1 (FEZF1-AS1) was found upregulated in colon cancer after screening all the lncRNAs of colon cancer tissues deposited in TCGA, the result of which was further confirmed by RNAscope staining on a colon tissue chip. The results obtained using FEZF1-AS1 knockout colon cancer cells (SW480 KO and HCT-116 KO) constructed using CRISPR/Cas9 system confirmed the proliferation, invasion, and migration-promoting function of FEZF1-AS1 in vitro. Mechanistically, FEZF1-AS1 associated with the mitochondrial protein phosphoenolpyruvate carboxykinase (PCK2), which plays an essential role in regulating energy metabolism in the mitochondria. Knockdown of FEZF1-AS1 greatly decreased PCK2 protein levels, broke the homeostasis of energy metabolism in the mitochondria, and inhibited proliferation, invasion, and migration of SW480 and HCT-116 cells. PCK2 overexpression in FEZF1-AS1 knockout cells partially rescued the tumor inhibitory effect on colon cancer cells both in vitro and in vivo. Moreover, PCK2 overexpression specifically rescued the abnormal accumulation of Flavin mononucleotide (FMN) and succinate, both of which play an important role in oxidative phosphorylation (OXPHOS). Overall, these results indicate that FEZF1-AS1 is an oncogene through regulating energy metabolism of the cell. This research reveals a new mechanism for lncRNAs to regulate colon cancer and provides a potential target for colon cancer diagnosis and treatment.

LATS1 exerts tumor suppressor functions via targeting Gli1 in colorectal cancer.

Background: The Hippo pathway's primary kinase component, large tumor suppressor 1 (LATS1), has been hypothesized as a tumor suppressor in a variety of cancers. LATS1's biological effects on colorectal cancer (CRC) are yet to be determined.Methods: The analysis of LATS1 mRNA expression in CRC was conducted using public databases from the Gene Expressing Profiling Interactive Analysis database (GEPIA). Investigation for the expression of LATS1 protein in 102 CRC tumor tissues and 57 normal tissues was performed using immunohistochemistry (IHC) analysis. In vitro genetic manipulation was used to explore the potential role and mechanism of LATS1 in the regulation of proliferation and migration of CRC cells.Results: LATS1 was found to be considerably downregulated in CRC tissues, with much lower levels in individuals with bigger tumors of size (≥5 cm), deeper invasion (T3-4), positive lymph node metastasis (LNM), and advanced tumor-node-metastasis (TNM) stage (III-IV). As exhibited by clinical data analysis, LATS1 loss was significantly associated with TNM and LNM staging in CRC patients. Furthermore, our in vitro investigations revealed that LATS1 depletion increased CRC cell proliferation and migration in HCT116 cells, whereas overexpressing LATS1 had the opposite effect in SW620 cells. LATS1 suppressed the expression of glioma-associated oncogene-1 (Gli1), and LATS1's tumor-suppressive actions in CRC are dependent on Gli1. Moreover, LATS1 could modulate Yes-associated protein 1 (YAP1) expression and mTOR activation in CRC cells.Conclusion: Our findings identify the LATS1 as a unique Gli1 regulator in CRC cell migration and proliferation, and suggest that LATS1 may serve as a potential therapeutic target for CRC.

Long noncoding RNA SNHG10 promotes colorectal cancer cells malignant progression by targeting miR-3690

ABSTRACT Long noncoding RNA small nucleolar RNA host gene 10 (SNHG10) has been suggested to function as tumor promoter in various human cancer types. Herein, the role of SNHG10 in colorectal cancer (CRC) was explored. Expression levels of genes in colorectal cancer tissues and cell lines were detected by Starbase and reverse transcription quantitative PCR (RT-qPCR) Cell Counting Kit-8 (CCK-8), the BrdU incorporation assay and Transwell assays were explored to study the function of SNHG10 in HCT116 and DXH-1 cells. In addition, the interaction of SNHG10 and miR-3690 was analyzed by dual-luciferase reporter assays. SNHG10 had a high expression level in CRC tissues and cell lines. Meanwhile, knockdown of SNHG10 reduced cell viability, inhibited cell proliferation and decreased cell migration and invasion. Moreover, bioinformatics analysis revealed that one potential target gene of SNHG10 was miR-3690. Dual-luciferase reporter assay confirmed that miR-3690 directly targeted SNHG10. Importantly, SNHG10 could decrease the expression of miR-3690 in HCT116 and DXH-1 cells. More importantly, the silencing of miR-3690 reversed the effect of the SNHG10 knockdown on the cell viability, proliferation, migration and invasion of HCT116 and DXH-1 cells. The present results demonstrated that SNHG10 promotes colorectal cancer cells the malignant progression by targeting miR-3690. Abbreviations: CRC: Colorectal cancer; Lnc RNA: Long noncoding RNA; microRNAs: miRNAs/miRs; RT-qPCR: reverse transcription quantitative polymerase chain reaction; CCK-8: Cell Counting Kit-8

Microbial Metabolite Deoxycholic Acid Promotes Vasculogenic Mimicry Formation in Intestinal Carcinogenesis

Background: High-fat diet (HFD) leads to exposure to gut microbial metabolite secondary bile acids such as deoxycholic acid (DCA) in the intestine, which is closely linked to colorectal cancer (CRC). Vasculogenic mimicry (VM) is a critical event for cancer malignant transformation. We aimed to investigate the crucial roles of DCA on VM and the intestinal carcinogenesis. Methods: The effects of HFD on VM formation and epithelial-mesenchymal transition (EMT) in human CRC tissues were investigated. The fecal DCA level was detected in the HFD treated- Apc min/+ mice. The four-week Apc min/+ mice were treated with 0·2% DCA for 12 weeks. Intestinal tumor tissues were collected for detection of VM, EMT, and vascular endothelial growth factor receptor 2 (VEGFR2) signaling. The effects of DCA on tube formation, EMT, and VEGFR2 signaling of HCT-116 cells were evaluated. Findings: Compared with the normal diet, HFD exacerbated VM formation and EMT in CRC patients. HFD could significantly increase fecal DCA level in Apc min/+ mice. More importantly, DCA promoted tumor cell proliferation and EMT, and increased VM formation accompanied by VEGFR2 activation, which led to intestinal carcinogenesis. DCA enhanced proliferation and migration of HCT-116 cells, and induced EMT and vitro tube formation. Furthermore, the silence of VEGFR2 blunted DCA-induced EMT, VM formation, and migration. Interpretation: Microbial metabolite DCA promoted VM formation and EMT through VEGFR2 activation, which further exacerbated intestinal carcinogenesis. Funding: This study was supported by the National Natural Science Foundation of China and the Key Project of Science and Technology Pillar Program of Tianjin. Declaration of Interest: The authors declare no conflicts of interest. Ethical Approval: This study was approved by the Ethics Committee of General Hospital, Tianjin Medical University, China.

FOXD3-induced miR-133a blocks progression and metastasis of colorectal cancer through regulating UBA2.

Background and Aim: Some studies have verified that miR-133a played an inhibitory role in several cancers. Whereas, the effect of miRNA-133a in colorectal cancer (CRC) has not been fully elucidated. Our study aims to confirm UBA2 as a direct target gene of miRNA-133a and explore the upstream modulatory molecules of miR-133a. In addition, their impacts on the biological characteristics of CRC cells were assessed.Methods: QRT-PCR analyzed miR-133a expression levels in colorectal cells including HCT116, SW48 cells and human normal colorectal cell line NCM460. A serial biological experiment assessed miR-133a effects on cell proliferation, migration, invasion and apoptosis capacities in HCT116 and SW48 cells. MiRNA targeting gene prediction and a dual luciferase assay were employed to confirm miR-133a-targeted UBA2. Transcription factors (TFs) FOXD3 was identified as an upstream regulator of miR-133a via JASPAR. The influence of miR-133a and FOXD3 on UBA2 expression was analyzed by qRT-PCR or western blot.Results: miR-133a was lowly expressed in CRC cells. High miRNA-133a expression suppressed the proliferation, migration, invasion and enhanced apoptosis capacities of CRC cells. MiR-133a targeted the UBA2 mRNA 3ʹUTR area and reduced UBA2 protein expression. We also unveiled that FOXD3 high-expression significantly raised miR-133a expression and diminished UBA2 expression. We also discovered that high miR-133a expression augmented the effects of elevated FOXD3 expression on CRC cell proliferation, migration and invasion, whereas, low miR-133a expression generated the opposite outcomes.Conclusion: FOXD3 induced miRNA-133a directly targeting UBA2 could affect the progression and growth of CRC.

Serum Starvation Sensitizes Anticancer Effect of Anemarrhena asphodeloides via p38/JNK-Induced Cell Cycle Arrest and Apoptosis in Colorectal Cancer Cells.

The molecular mechanism underlying the anticancer effects of Anemarrhena asphodeloides (A. asphodeloides) on colon cancer is unknown. This is the first study evaluating the anticancer effect of A. asphodeloides extract (AA-Ex) in serum-starved colorectal cancer cells. Changes in cell proliferation and morphology in serum-starved MC38 and HCT116 colorectal cancer cells were investigated using MTS assay. Cell cycle and apoptosis were investigated using flow cytometry, and cell cycle regulator expression was determined using qRT-PCR. Apoptosis regulator protein levels and mitogen-activated protein kinase (MAPK) phosphorylation were assessed using western blotting. AA-Ex sensitively suppressed proliferation of serum-starved colorectal cancer cells, with MC38 and HCT116 cells showing greater changes in proliferation after treatment with AA-Ex under serum starvation than HaCaT and RAW 264.7 cells. AA-Ex inhibited cell cycle progression in serum-starved MC38 and HCT116 cells and increased the expression of cell cycle inhibitors (p53, p21, and p27). Furthermore, AA-Ex induced apoptosis in serum-starved MC38 and HCT116 cells. Consistently, AA-Ex suppressed the expression of the anti-apoptotic molecule Bcl-2 and upregulated pro-apoptotic molecules (cytochrome c, cleaved caspase-9, cleaved caspase-3, and cleaved-PARP) in serum-starved cells. AA-Ex treatment under serum starvation decreased AKT and ERK1/2 phosphorylation in the cell survival signaling pathway but increased p38 and JNK phosphorylation. Furthermore, AA-Ex treatment with serum starvation increased the levels of the transcription factors of the p38 and JNK pathway. Serum starvation sensitizes colorectal cancer cells to the anticancer effect of A. asphodeloidesvia p38/JNK-induced cell cycle arrest and apoptosis. Hence, AA-Ex possesses therapeutic potential for colon cancer treatment.

SPOCK1 promotes the proliferation and migration of colon cancer cells by regulating the NF-κB pathway and inducing EMT.

Sparc/osteonectin, cwcv, and kazal-like domains proteoglycan 1 (SPOCK1) has been shown to promote various tumors, but its role in colon cancer (CRC) has not been clearly illuminated. The aim of this study was to investigate the effects of SPOCK1 interference on the proliferation, migration, and EMT of CRC cells. First, we analyzed the expression of SPOCK1 in various CRC datasets. Then, we investigated the correlation between SPOCK1 and prognosis in CRC patients. We overexpressed SPOCK1 and knocked down SPOCK1 expression in HCT-116 and SW480 cells, respectively. Then, cell proliferation was assayed with a CCK-8 assay, and cell migration was evaluated with a Transwell migration assay. NF-κB and EMT-related proteins were studied by western blotting. The results indicated that the mRNA levels of SPOCK1 were relatively high in CRC tissues and that high expression of SPOCK1 was negatively correlated with patient prognosis. With SPOCK1 overexpression in HCT-116 cells, cell proliferation and migration were increased, while SPOCK1 knockdown had the opposite effects. With SPOCK1 overexpression in HCT-116 cells, the expression levels of NF-κB and EMT-related proteins were elevated, while SPOCK1 knockdown produced the opposite results. In conclusion, our study demonstrates that SPOCK1 may activate the NF-κB/Snail signaling cascade to promote the proliferation and migration of CRC cells. SPOCK1 may serve as a new prognostic indicator and potential therapeutic target in CRC.

Lysosome-Mediated Cytotoxic Autophagy Contributes to Tea Polysaccharide-Induced Colon Cancer Cell Death via mTOR-TFEB Signaling.

Targeting autophagy and lysosome may serve as a promising strategy for cancer therapy. Tea polysaccharide (TP) has shown promising antitumor effects. However, its mechanism remains elusive. Here, TP was found to have a significant inhibitory effect on the proliferation of colon cancer line HCT116 cells. RNA-seq analysis showed that TP upregulated autophagy and lysosome signal pathways, which was further confirmed through experiments. Immunofluorescence experiments indicated that TP activated transcription factor EB (TFEB), a key nuclear transcription factor modulating autophagy and lysosome biogenesis. In addition, TP inhibited the activity of mTOR, while it increased the expression of Lamp1. Furthermore, TP ameliorated the lysosomal damage and autophagy flux barrier caused by Baf A1 (lysosome inhibitor). Hence, our data suggested that TP repressed the proliferation of HCT116 cells by targeting lysosome to induce cytotoxic autophagy, which might be achieved through mTOR-TFEB signaling. In summary, TP may be used as a potential drug to overcome colon cancer.

Photoprotective, antioxidant, antimicrobial and cytotoxic activities of guarana (Paullinia cupana) seed extracts

Introduction: Guarana (Paullinia cupana) is a native Amazonian fruit and, due to its medicinal properties, stands out as one of the most promising species of Brazilian flora. Known for its invigorating and stimulating action, this species produces phenolic compounds responsible for its biological activities, which are of great interest to the phytocosmetic industry, in the preparation of anti-aging creams, for example. In this study, the antimicrobial, antioxidant, photoprotective and cytotoxic activities of extracts obtained from guarana seeds were evaluated. Methods: Guarana seeds were obtained in Maués, Amazonas state, Brazil and, after drying, the seeds were ground in a knife mill. P. cupana seed extracts were obtained by the depletion method using hexane, dichloromethane and ethanol as solvents. Antioxidant activity assays were performed to determine the ability to scavenge the free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH•). To assess the sun protection factor (SPF), Mansur's spectrophotometric method was used. For the analysis of antimicrobial activity, the microplate dilution technique was used against strains of Escherichia coli, Pseudomonas aeruginosa and Candida albicans. To evaluate the toxicity of the extracts, the lethal concentration of 50% of the population of the microcrustacean Artemia salina was determined. The cytotoxic effect of the extracts against human colorectal carcinoma cells (HCT 116) was also evaluated using the Alamar Blue test. Results: The ethanolic extract from guarana seeds showed SPF value of 249.10 and a critical wavelength number (λc) of 383 nm, when evaluated at a concentration of 0.1% (m/m). The ethanolic extract also showed antioxidant activity, and a 50% efficient concentration (EC50) of 647 μg mL-1. The dichloromethane extract was active against E. coli and P. aeruginosa, with a minimum inhibitory concentration (MIC) of 20 mg mL-1 for both bacteria, and a MIC of 2.0 mg mL-1 for the yeast. This extract did not show toxicity against A. salina (LD50> 1 mg mL-1). The ethanolic and hexane extracts, however, presented low toxicity to the microcrustacean. On the other hand, none of the evaluated extracts inhibited the proliferation of HCT116 cells, showing no cytotoxicity. Conclusion: P. cupana seeds have an important therapeutic value due to the presence of substances with antioxidant, antibiotic and photoprotective activities, though with low cytotoxicity, which allows their use as functional additives in phytocosmetics, as well as in the development of possible phytotherapeutic products.

S100A16 suppresses the proliferation, migration and invasion of colorectal cancer cells in part via the JNK/p38 MAPK pathway.

S100 calcium binding protein A16 (S100A16) is the most recent member of the S100 calcium-binding protein family. The function of S100A16 has been associated with various types of cancer; however, its role in colorectal cancer (CRC) remains unknown. Therefore, the aim of the present study was to investigate the role of S100A16 in CRC progression. The Oncomine dataset used in the current study revealed that the expression of S100A16 was decreased in CRC compared with normal colorectal tissues. Similar results were also determined via immunohistochemistry. In addition, a negative association was identified between S100A16 expression and the prognosis of patients with CRC. Further functional experiments revealed that S100A16 knockdown promoted the proliferation, migration and invasion of HCT116 and SW480 cells, and vice versa in Lovo cells. Epithelial-mesenchymal transition (EMT) was promoted and the JNK/p38 MAPK pathway was activated in HCT116 cells following S100A16 knockdown, as determined via western blotting. Furthermore, S100A16 silencing promoted the migration and invasion of cells. EMT was also reversed when cells were treated with the JNK inhibitor (SP600125) or the p38 inhibitor (SB203580). In summary, the results of the present study demonstrated that S100A16 suppressed the proliferation, migration and invasion of CRC cells partially via the JNK/p38 MAPK signalling pathway and subsequent EMT mediation.

MicroRNA‑223‑induced inhibition of the FBXW7 gene affects the proliferation and apoptosis of colorectal cancer cells via the Notch and Akt/mTOR pathways.

The tumour suppressor gene F-box and WD repeat domain-containing 7 (FBXW7) plays an important role in human cancer by regulating cell division, proliferation and differentiation. However, the exact regulatory mechanisms of microRNA (miR)-223 in colorectal cancer (CRC) cells are still unknown. The present study aimed to investigate the effect and mechanism of miR-223 inhibiting FBXW7 on the proliferation and apoptosis of CRC cells. HCT116 cells were transfected with miR-223 mimics or small interfering RNA (siRNA) targeting FBXW7 (siFBXW7), and the effects of these treatments on cell proliferation and apoptosis were examined. The downstream Notch and Akt/mTOR pathways were also assessed. Following miR-223 overexpression, the mRNA and protein expression levels of FBXW7 were downregulated. Transfection with miR-223 mimics or siFBXW7 promoted the proliferation of HCT116 cells and inhibited apoptosis by promoting the Notch and Akt/mTOR signalling pathways. Conversely, miR-223 mimics transfection with FBXW7 overexpression inhibited cell viability and restored apoptosis. Thus, the present study demonstrated that miR-223 could bind to the FBXW7 gene and inhibit its expression, ultimately increasing the proliferation and preventing the apoptosis of CRC cells through the Notch and Akt/mTOR signalling pathways.

Matrine Can Inhibit the Growth of Colorectal Cancer Cells by Inducing Ferroptosis

Colorectal cancer (CRC) is a common malignant tumor of the digestive system that can seriously threaten human health. Chinese matrine is known to have a wide range of antiviral and immunomodulatory effects. In this study, we evaluated the effect of matrine on ferroptosis using the HCT116 human colon cancer cell line. We evaluated cell viability and proliferation using the cell counting kit-8 assay and carried out cell clone formation experiment by measuring reactive oxygen species (ROS) production, as well as levels of glutathione (GSH), lipid peroxide (MDA), Fe2+, glutathione peroxidase 4 (GPX4), activating transcription factor 4 (ATF4), solute carrier family 7 member 11 (SLC7A11), transferrin receptor protein 2 (TFR2), and Sigma-1 receptor (Sigma-1R) in order to evaluate the redox status of cells. These results indicate that matrine can significantly reduce the cell viability of HCT116 and decrease cell proliferation. After treatment with matrine, ROS, Fe2+, and MDA levels increased significantly, while the GSH content decreased. In addition, the expression of GPX4, SLC7A11, and Sigma-1R decreased significantly, while the expression of ATF4 and TFR2 increased significantly. These results indicate that matrine can induce ferroptosis in CRC, which can provide new clues for further pharmacological research of matrine and provide experimental evidence for understanding its mechanism of inhibiting CRC.