Proliferation Of Colon Cancer Cells Research Articles

Potential bioactivities via anticancer, antioxidant, and immunomodulatory properties of cultured mycelial enriched β-D-glucan polysaccharides from a novel fungus Ophiocordyceps sinensis OS8.

Ophiocordyceps sinensis is well-known worldwide as a traditional medicine. An alternative natural source of O. sinensis is provided by mycelial cultivation. However, the bioactivities of cultured mycelial-enriched β-D-glucan polysaccharides from a novel fungus O. sinensis OS8 are still unknown. We investigated the potential bioactivities via anticancer, antioxidant, and immunomodulatory polysaccharides (OS8P) produced from cultured mycelia of O. sinensis OS8. This strain is a novel fungus isolated from natural O. sinensis, which is further cultured by submerged mycelial cultivation for polysaccharide production. The yield of mycelial biomass was 23.61 g/l, and it contained 306.1 mg/100 g of adenosine and 3.22 g/100 g of polysaccharides. This OS8P was enriched with β-D-glucan at 56.92% and another form of α-D-glucan at 35.32%. The main components of OS8P were dodecamethyl pentasiloxane, 2,6-bis (methylthiomethyl) pyridine, 2-(4-pyrimidinyl)-1H-Benzimidazole, and 2-Chloro-4-(4-nitroanilino)-6-(O-toluidino)-1,3,5-triazine at the rates of 32.5, 20.0, 17.5, and 16.25%, respectively. The growth of colon cancer cells (HT-29) was significantly inhibited by OS8P, with IC50 value of 202.98 µg/ml, and encouraged apoptosis in HT-29 cells as confirmed by morphological change analysis via AO/PI and DAPI staining, DNA fragmentation, and scanning electron microscopic observations. In addition, significant antioxidant activity was demonstrated by OS8P through DPPH and ABTS assays, with IC50 values of 0.52 and 2.07 mg/ml, respectively. The OS8P also exhibited suitable immunomodulatory activities that significantly enhanced (P< 0.05) the induction of splenocyte proliferation. The OS8P enriched with β-D-glucan polysaccharides and produced by submerged mycelial culture of a new fungal strain of O. sinensis OS8 strongly inhibited the proliferation of colon cancer cells without any cytotoxicity against normal cells. The potential effect of the OS8P on the cancer cells was due to the stimulation of apoptosis. Also, the OS8P exhibited good antioxidant and immunomodulatory activities. The results indicate that OS8P has promising applications in the functional food industry and/or therapeutic agents for colon cancer.

Overexpression of hBD3 inhibits cell proliferation, cell cycle, and migration in colon cancer cells

Overexpression of hBD3 inhibits cell proliferation, cell cycle, and migration in colon cancer cells

Abstract 3699: Involvement of p53 and KRAS in the regulation of SLC38A5 in colon cancer

Abstract Cancer-associated upregulation has been demonstrated for selective glucose transporters and amino acid transporters. The amino acid transporter SN2 (SLC38A5) represents the latest addition to this list. Transcriptomic analysis of tumor tissues has shown SLC38A5 is upregulated in breast and colon cancer. SLC38A5 is an amino acid-dependent Na+/H+ exchanger, induced in cancer, which not only supplies amino acids to cancer cells but also maintains an alkaline intracellular pH. The substrates for SLC38A5 include glutamine, asparagine, histidine, methionine, glycine, and serine, highlighting the role of SLC38A5 in glutamine addiction and one-carbon metabolism, both pathways being essential for cancer cells. In addition, SLC38A5 activates macropinocytosis, a process involved in cellular uptake of proteins in the extracellular fluid to meet amino acid demands in cancer cells. Since the transporter is upregulated in colon cancer, we hypothesized that the multiple functions of SLC38A5 fuel the growth, proliferation, and survival in colon cancer cells and that its induction involves oncogenic mutations in p53 and KRAS that are common mediators of carcinogenesis in colon. Firstly, we used a panel of colon cancer cells and analyzed SLC38A5 expression and its functional characteristics. Next, we compared SLC38A5 expression and function between two isogenic cell lines: SW48 with and without the oncogenic mutation (G12D) in KRAS and HCT116 with and without the tumor suppressor p53. Among the colon cancer cells examined, KM12L4 showed highest expression of SLC38A5; therefore, we characterized the function of the transporter in detail in this cell line. Our studies showed that KM12L4 cells were able to take up serine, one of the substrates for SLC38A5, in a Na+-dependent manner in the presence of 5 mM tryptophan, which is not a substrate for SLC38A5 but is known to block LAT1 transporter, which also transports serine. SLC38A5 exhibited an active serine-uptake when Na+ was replaced with Li+ at a higher pH. Li+ tolerance and a higher activity at an alkaline pH are two unique features of SLC38A5. The oncogenic mutation G12D in KRAS increased SLC38A5 activity in SW48 cells. In contrast, loss of p53 decreased SLC38A5 activity in HCT116 cells. These data demonstrate that SLC38A5 is induced in colon cancer cells and its activity is increased in the presence of oncogenic KRAS mutations and suppressed when p53 is inactivated, thus providing a valuable insight into the molecular mechanisms for the upregulation of the transporter in colon cancer. We conclude that SLC38A5, an amino acids transporter, is upregulated in colon cancer and its functions are uniquely suited to promote cell proliferation and tumor growth. We also conclude that KRAS and p53 functional status play a critical role in the increased SLC38A5 expression and activity in colon cancer. These data suggest that SLC38A5 could be exploited as a drug target for cancer therapy. Citation Format: Nhi Thi Nguyen, Sathish Sivaprakasam, Vadivel Ganapathy. Involvement of p53 and KRAS in the regulation of SLC38A5 in colon cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3699.

Carboxypeptidase A6 suppresses the proliferation and invasion of colorectal cancer cells and is negatively regulated by miR-96–3p

BackgroundColorectal cancer (CRC) is a common malignant tumor, and this study aims to explore the role and the regulatory mechanism of carboxypeptidase A6 (CPA6) in CRC cells. MethodsSpecific shRNA targeting CPA6 mRNA was transfected into NCM460 and HT29 cells to down-regulate CPA expression, and expression plasmid was transfected into HCT116 cells to exogenously overexpress CPA6. The dual luciferase assay was used to detect the direct binding of miR-96–3p to CPA6 3′UTR. Phosphorylation and activation of Akt were detected using Western blot. Cells were treated with miR-96–3p mimics, Akt inhibitor (MK-2206) or agonist (SC79) for rescue experiments. The cell functions were evaluated using CCK-8, clone formation, transwell, and Western blot assays. Xenograft tumor assay was also used to analyze the effect of altered CPA6 expression on tumor growth. ResultsKnockdown of CPA6 promoted the proliferation, clone formation, migration, and invasion of NCM460 and HT29 cells in vitro, and the tumor growth of nude mouse xenograft tumor in vivo. Moreover, over-expression of CPA6 significantly inhibited the malignant proliferation and invasion of HCT116 cells in vitro, and the tumor growth of xenograft tumor in vivo. Furthermore, miR-96–3p could directly regulate CPA6 expression by targeting its 3′UTR, and miR-96–3p mimics rescued the inhibitory effects of CPA6 overexpression on the malignant proliferation and invasion of CRC cells. Finally, CPA6 knockdown enhanced Akt/mTOR phosphorylation and activation, while CPA6 overexpression inhibited Akt/mTOR activation. The regulatory effect of CPA6 on Akt/mTOR signaling was naturally regulated by miR-96–3p. Akt inhibitor or agonist rescued the effects of CPA6 knockdown or overexpression on proliferation and EMT of colon cancer cells. ConclusionCPA6 has a significant tumor suppressive effect on CRC by inhibiting the activation of Akt/mTOR signaling, and miR-96–3p negatively regulates the expression of CPA6.

Lokal anesteziklerden levobupivakain ve ropivakain kolon kanseri tedavisinde kullanılabilir mi?

Purpose: Colon cancer is among the most common causes of death among cancers, and in the treatment of colon cancer, adjuvant chemotherapy is applied mainly after curative surgery, to prevent recurrence and metastases after resection. In recent years, local anesthetics have received increasing attention in cancer research. This study aimed to investigate the proliferative and apoptotic effects of local anesthetics levobupivacaine and ropivacaine on SW480 colon cancer cells.&#x0D; Materials and Methods: In the study, different concentrations and durations of levobupivacaine and ropivacaine were applied to the SW480 colon cancer cell line and their effect on proliferation was determined by MTT analysis. The TUNEL method was used to determine its apoptotic activity. The results were evaluated statistically.&#x0D; Results: It was determined that levobupivacaine and ropivacaine decreased cell proliferation in SW480 colon cancer cells depending on dose and time. It was also observed to promote apoptosis in colon cancer cells.&#x0D; Conclusion: According to the study data, it was determined that levobupivacaine and ropivacaine acted cytotoxic by activating apoptosis in SW480 colon cancer cells. It is thought that these data will lead to multidisciplinary studies to elucidate the antitumor effect mechanism of local anesthetics.

YWHAB knockdown inhibits cell proliferation whilst promoting cell cycle arrest and apoptosis in colon cancer cells through PIK3R2.

Colon cancer is one of the most common causes of cancer-associated mortality. Tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein β (YWHAB) has been reported to be aberrantly expressed in human colon cancer cells following alltrans retinoic acid treatment. PI3K regulatory subunit 2 (PIK3R2) has also been identified as a gene associated with colon cancer metastasis and tumor progression. The present study aimed to determine the role of YWHAB in colon cancer in addition to its detailed reaction mechanism. The expression levels of YWHAB and PIK3 'A0* R2 before or after transfection of YWHAB interference plasmids or PIK3R2 overexpression plasmids were examined by reverse transcription-quantitative PCR and western blotting. PI flow cytometry, Cell Counting Kit-8 and TUNEL assays were performed to measure the extent of cell cycle progression, proliferation and apoptosis. Additionally, the expression levels of G1-S cell-cycle transition regulator cyclin D1 and G1-checkpoint CDK inhibitor p21 and apoptosis marker proteins Bcl2 and Bax were assessed using western blotting. Subsequently, the Monarch Initiative database (https://monarchinitiative.org/) predicted the binding of YWHAB and PIK3R2, following which co-immunoprecipitation assay was utilized to assess their potential interaction. Furthermore, western blotting was performed to examine the expression levels of PI3K/AKT signaling pathway markers. It was revealed that YWHAB expression was upregulated in colon cancer cells compared with HIEC-6 human intestinal epithelial cells. Functionally, YWHAB depletion by transfection of YWHAB interference plasmids was demonstrated to suppress the proliferation of colon cancer cells whilst promoting cell cycle arrest at the G0/G1 phase and apoptosis, decreasing cyclin D1 and Bcl2 expression, and increasing p21 and Bax expression. Additionally, YWHAB was verified to bind to PIK3R2 and YWHAB knockdown decreased PIK3R2 expression. PIK3R2 overexpression by transfection of PIK3R2 overexpression plasmids reversed the effects of YWHAB knockdown on cell proliferation, cycle arrest, apoptosis, and apoptotic and cell cycle proteins in colon cancer cells. YWHAB knockdown reduced the levels of p-PI3K/PI3K and p-AKT/AKT and PIK3R2 overexpression also reversed the effects of YWHAB knockdown on the expression of PI3K/AKT signaling markers. In conclusion, these data suggest that YWHAB can activate the PI3K/AKT signaling pathway and participate in the malignant progression of colon cancer by targeting PIK3R2, which provides novel insights into the mechanism of colon cancer.

CHRM3-Associated miRNAs May Play a Role in Bile Acid-Induced Proliferation of H508 Colon Cancer Cells.

It was well defined that proliferative effects of bile acids on colon epithelium are through interaction with muscarinic-3 receptors. Recently, microRNA emerged as an important regulator of gene expression and has been implicated in pathogenesis of many malignancies. However, the interaction of CHRM3 and microRNAs and their potential effects on colon carcinogenesis remains to be elucidated. In the current study, analysis of cell proliferation for 6 days after treatment with sodium taurolithocholate was analyzed by using WST-1 method. microRNAs which possibly target CHRM3 were identified by in silico analyses. Expression profiling of these microRNAs, expression changes of CHRM3 gene at mRNA level for H508 and SNU-C4 colon cancer cells were analyzed by quantitative polymerase chain reaction; the protein level of CHRM3 was analyzed using Western blot; apoptotic experiments were analyzed using the Annexin V assay. The Gene Ontology and the Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed using the miRPath v3.0. It was found that the expression level of CHRM3 gene was 6.133 ± 0.698-fold in H508 cells compared with SNU-C4 cells (P =.004). Treatment of H508 cells with sodium taurolithocholate caused 1.34 ± 0.4156-fold change in the expression level of CHRM3 gene (P =.0448). No apoptotic changes were observed in both colon cancer cells after treatment with sodium taurolithocholate. Different expression changes were detected of hsa-miR-129-5p, hsa-miR-30c-5p, hsa-miR-224-5p, hsa-miR-30b-5p, hsa-miR-522-3p, and hsa-miR-1246. Finally, hsa-miR-1246 and hsa-miR-522-3p could play a critical role in tumor development via bile acid-related genes in colon cancer. These findings reflected that CHRM3-dependent oncogenetic pathways might be in charge of colon cancer. We suggest that the microRNA expression profile of each individual colon cancer tissue is a unique digital signature.

SETD8 promotes glycolysis in colorectal cancer via regulating HIF1α/HK2 axis

Glycolysis is one of the factors influencing cancer cell growth and metastasis. Here, we aimed to investigate the role of SETD8 gene, which is a pro-oncogene. Using bioinformatics tools including Ualcan, Timer, GEPIA, and PrognoScan to study the expression of SETD8 in colorectal cancer, we found that SETD8 expression was higher in colon cancer tissues than that in normal tissues. Higher levels of SETD8 predicted poorer survival of patients. This piqued our interest, so we transfected SETD8 knockdown and overexpression plasmids into colorectal cancer cells and found that SETD8 overexpression enhanced proliferation and glycolysis in colon cancer cells, while SETD8 knockdown decreased cell proliferation and glycolysis. Mechanistically, we examined the expression of HIF1α and HK2 protein by western-blot assay and found that SETD8 activated the HIF1α/HK2 pathway. Then, we treated SETD8-overexpressed cells with HIF1α inhibitor and found that the pro-tumor growth and glycolytic effects of SETD8 were reversed, indicating that SETD8 promoted the growths of colorectal cancer cells by upregulating the HIF1α /HK2 pathway.

Phloretin induces G2/M arrest and apoptosis by suppressing the β-catenin signaling pathway in colorectal carcinoma cells.

Colorectal carcinoma (CRC) is the third most prevalent cancer, causing a significant mortality worldwide. Present available therapies are surgery, chemotherapy including radiotherapy, and these are known to be associated with heavy side effects. Therefore, nutritional intervention in the form of natural polyphenols has been well recognised to prevent CRC. Phloretin, a known dihydrochalcone is present in apple, pear and strawberry. This has been proven to induce apoptosis in cancer cells and also exhibited anti-inflammatory activity, thus can be explored as a potential anticancer nutraceutical agent. This study demonstrated phloretin's significant in vitro anticancer activity against CRC. Phloretin suppressed cell proliferation, colony forming ability and cellular migration in human colorectal cancer HCT-116 and SW-480 cells. Results also revealed that phloretin generated reactive oxygen species (ROS) which provoked depolarization of mitochondrial membrane potential (MMP) and further contributed to cytotoxicity in colon cancer cells. Phloretin also modulated the cell cycle regulators including cyclins and cyclin-dependent kinases (CDKs) and halted cell cycle at G2/M phase. Moreover, it also induced apoptosis by regulating the expression of Bax and BCl-2. The Wnt/β-catenin signaling is inactivated by phloretin by targeting the downstream oncogenes namely CyclinD1, c-Myc and Survivin which are involved in the proliferation and apoptosis of colon cancer cells. In our study we showed that lithium chloride (LiCl) induced the expression of β-catenin and its target genes and the co-treatment of phloretin circumvent its effect and downregulated the Wnt/β-catenin signaling. In conclusion, our results strongly suggested that phloretin can be utilized as a nutraceutical anticancer agent for combating CRC.

Probiotic Properties of Loigolactobacillus coryniformis NA-3 and In Vitro Comparative Evaluation of Live and Heat-Killed Cells for Antioxidant, Anticancer and Immunoregulatory Activities.

Some Latiactobacilli are often used as probiotics due to their functional activities, including antioxidant, anticancer and immunoregulation effect. Loigolactobacillus coryniformis NA-3 obtained from our laboratory is a promising probiotic according to the previous study. Coculture, the Oxford cup test and disk-diffusion methods were used to evaluate the probiotic properties and antibiotic resistance of L. coryniformis NA-3. The antioxidant activities of live and heat-killed L. coryniformis NA-3 were assessed via radicals' scavenging ability. The potential anticancer and immunoregulatory capacity was determined in vitro using cell lines. The results indicate that L. coryniformis NA-3 has antibacterial activity and cholesterol removal ability and is sensitive to most antibiotics. Dead L. coryniformis NA-3 can scavenge free radicals as well as live strains. Live L. coryniformis NA-3 can significantly inhibit the proliferation of colon cancer cells; however, dead cells cannot. After RAW 264.7 macrophages were treated with live and heat-killed L. coryniformis NA-3, the production of NO, IL-6, TNF-α and reactive oxygen species (ROS) was induced. The increased expression of inducible nitric oxide synthase (iNOS) in treated macrophages mediates the production of NO. In conclusion, L. coryniformis NA-3 showed potential probiotic properties, and the heat-killed strain also exhibited activities similar to those of live bacteria, suggesting the possible value of its further application in the food processing and pharmaceutical industries.

Gelsolin Inhibits the Proliferation of Colon Cancer Cells by Enhancing the Expression of TNFR2/CASP10 as a Death Receptor Pathway.

Colon cancer has the second highest incidence rate of digestive system tumors. It relies on surgical treatment, radiotherapy and chemotherapy, and targeted drug therapy. To study the mechanism of GSN in the proliferation of colon cancer cells. The expression of gelsolin (GSN) was analyzed with the data of colon cancer patients in the TCGA database. SW620 cells were treated by GSN in vitro and the gene expression was detected by immunoblotting and quantitative PCR. The expression of GSN was found significantly low in colon cancer cells and correlated with the prognosis of patients. The SW620 cell line cultured in vitro was treated with exogenous GSN. SW620 can be significantly inhibited above the concentration of 250 μg/ml. The results of immunoblotting and quantitative PCR showed that exogenous GSN can effectively improve the transcription level of death receptor-related pathway genes such as TNFR2 and CASP10. This study found that GSN inhibited the proliferation of SW620 cells in vitro by upregulating the expression of death receptor pathway-related proteins.

Aerobic glycolysis in colon cancer is repressed by naringin via the HIF1Α pathway.

Metabolic reprogramming is a common phenomenon in cancer, with aerobic glycolysis being one of its important characteristics. Hypoxia-inducible factor-1α (HIF1Α) is thought to play an important role in aerobic glycolysis. Meanwhile, naringin is a natural flavanone glycoside derived from grapefruits and many other citrus fruits. In this work, we identified glycolytic genes related to HIF1Α by analyzing the colon cancer database. The analysis of extracellular acidification rate and cell function verified the regulatory effects of HIF1Α overexpression on glycolysis, and the proliferation and migration of colon cancer cells. Moreover, naringin was used as an inhibitor of colon cancer cells to illustrate its effect on HIF1Α function. The results showed that the HIF1Α and enolase 2 (ENO2) levels in colon cancer tissues were highly correlated, and their high expression indicated a poor prognosis for colon cancer patients. Mechanistically, HIF1Α directly binds to the DNA promoter region and upregulates the transcription of ENO2; ectopic expression of ENO2 increased aerobic glycolysis in colon cancer cells. Most importantly, we found that the appropriate concentration of naringin inhibited the transcriptional activity of HIF1Α, which in turn decreased aerobic glycolysis in colon cancer cells. Generally, naringin reduces glycolysis in colon cancer cells by reducing the transcriptional activity of HIF1Α and the proliferation and invasion of colon cancer cells. This study helps to elucidate the relationship between colon cancer progression and glucose metabolism, and demonstrates the efficacy of naringin in the treatment of colon cancer.

G-quadruplex-enhanced circular single-stranded DNA (G4-CSSD) adsorption of miRNA to inhibit colon cancer progression.

Chromosomal heterogeneity leads to the abnormal expression and mutation of tumor-specific genes. Drugs targeting oncogenes have been extensively developed. However, given the random mutation of tumor suppressor genes, the development of its targeted drugs is difficult. Our early research revealed that artificial circular single-stranded DNA (CSSD) can restore multiple tumor suppressor genes to inhibit tumor malignant progression by adsorbing miRNA. Here, we improved CSSD to a fully closed single-stranded DNA with G quadruplex DNA secondary structure (G4-CSSD), which made G4-CSSD with higher acquisition rate and decreased degradation. The Cancer Genome Atlas (TCGA) and Human Protein Atlas database were used to predict tumour suppressor genes in colon cancer. Cellular and animal experiments were performed to validate the role of G4-CSSD in cancer cell progression. In colon cancer, we observed the simultaneous low expressions of chloride channel accessory 1 (CLCA1), UDP-GlcNAc:betaGal beta-1,3-N-acetylglucosaminyltransferase 6 (B3GNT6) and UDP glucuronosyltransferase family 2 member A3 (UGT2A3), which indicated an favourable prognosis. After repressing miR-590-3p with G4-CSSD590, the upregulation of CLCA1, B3GNT6 and UGT2A3 inhibited the proliferation and metastasis of colon cancer cells. This study may provide basis for new treatment methods for colon cancer by restoration of tumor suppressor genes.

XBP1s acts as a transcription factor of IRE1α and promotes proliferation of colon cancer cells

Upon ER stress, IRE1α is activated to splice XBP1 mRNA to generate XBP1s, a transcription factor that induces the expression of genes to cope with the stress. Expression of IRE1α is elevated in cancers and the IRE1α-XBP1s axis plays an important role in proliferation of cancer cells. However, the underlying mechanism is not well known. We found that ER stressors induced the expression of IRE1α, which was inhibited by depletion of XBP1s. XBP1s bound IRE1α promoter and initiated the transcription of IRE1α. These data indicate that XBP1s acts as a transcription factor of IRE1α. Overexpression of XBP1s increased the phosphorylation of JNK, a substrate of IRE1α kinase, which was inhibited by IRE1α kinase inhibitor Kira8. Overexpression of XBP1s also activated the regulated IRE1-dependent decay of mRNAs, which was suppressed by IRE1α RNase inhibitor STF083010. Moreover, we found that expression of XBP1s promoted proliferation of colon cancer cells, which was abrogated by Kira8 and STF083010. The results suggest that XBP1s functions to induce IRE1α expression and promote cancer cell proliferation. Our findings reveal a previously unknown mechanism of IRE1α expression by XBP1s and highlight the role of this regulation in proliferation of colon cancer cells, suggesting that IRE1α-targeting is a potential therapeutic strategy for colon cancer.

A Comprehensive Study on the Anti-cancer Effects of Quercetin and Its Epigenetic Modifications in Arresting Progression of Colon Cancer Cell Proliferation

Colon cancer etiology involves a wide spectrum of genetic and epigenetic alterations, finding it challenging to find effective therapeutic strategies. Quercetin exhibits potent anti-proliferative/apoptotic properties. In the present study, we aimed to elucidate the anti-cancer and anti-aging effect of quercetin in colon cancer cell lines. The anti-proliferative effect of quercetin was assessed in vitro by CCK-8 in normal and colon cancer cell lines. To check the anti-aging potential of quercetin, collagenase, elastase, and hyaluronidase inhibitory activity assays were performed. The epigenetic and DNA damage assays were performed using the human NAD-dependent deacetylase Sirtuin-6, proteasome 20S, Klotho, Cytochrome-C, and telomerase ELISA kits. Furthermore, the aging-associated miRNA expression profiling was performed on colon cancer cells. The treatment with quercetin inhibited cell proliferation of colon cancer cells in a dose-dependent manner. Quercetin arrested colon cancer cell growth by modulating expression of aging proteins including Sirtuin-6 and Klotho and also by inhibiting telomerase activity to restrict the telomere length which is evident from qPCR analysis. Quercetin also exhibited DNA damage protection by reducing proteasome 20S levels. The miRNA expression profiling results displayed differential expression of miRNA in colon cancer cell, and in addition, the highly upregulated miRNA was involved in the regulation of cell cycle, proliferation, and transcription. Our data suggest that quercetin treatment inhibited cell proliferation in colon cancer cells through regulating the anti-aging protein expression and provides better understanding for quercetin’s potential use in colon cancer treatment.

Prevention of Cell Proliferation Signaling Via RNA Interference of c-myc Gene Expression in Colon Cancer In-vitro

Colon cancer is a cancer begins in the large intestine (colon). Its aggression is due to late diagnosis, so poor prognosis and higher mortality rates are reported. Colon cancer has become a thorny research area that requires more examination of cellular pathways involved in its emergency. Based on this, we sought to check the biological role of cMyc in colon cancer cell proliferation through targeting its expression by a respective siRNA using CaCo-2 cell lines. Notably, the cytotoxic potential of cMyc knockdown was monitored by cell imaging using an inverted microscope, the production of lactate dehydrogenase (LDH) and viability of the transfected cells using MTT assay. The relative gene expression of c-myc and other related factors such as Raf-1 was investigated using quantitative real-time PCR (qRT-PCR). ELISA assay has been used to achieve the production of pro and anti-inflammatory cytokines released from transfected cells. Interestingly, the transfection with a new designed siRNA antagonist cMyc markedly decreased CaCo-2 cell viability rate in a dose-dependent manner compared with the transfection with specific siRNA antagonist Luciferase gene, which served as control. Likewise, number of survived cells significantly reduced following transfection with siRNA against c-myc, while the relative production of LDH significantly increased. Importantly, the cytotoxic effect of the transfection protocol showed neglected influence in normal colon epithelial cells (NCM-460 cell lines) indicating the selective regulation of colon cancer cell proliferation by transfection with siRNA targeting c-myc. Noteworthy, the knockdown efficiency of c-myc showed more than 80% inhibition of its expression in CaCo-2 cells transfected with the siRNA antagonist c-myc indicated by qRT-PCR. The relative gene expression of Raf-1 significantly decreased in c-myc knockdown cells, while the relative expression of both P53 and Caspase 3, as cell death indicators, significantly upregulated compared with control-transfected cells. Furthermore, the transfection with siRNA targeting c-myc markedly increased the production of interleukin 1 alpha (IL-1α ) and IL-1β, as pro-inflammatory cytokines, while decreased the production of IL- 4 and IL-10, as anti-inflammatory cytokines. These data provide evidence for the involvement of c-myc gene expression in colon cancer cell proliferation and immune response during cancer development.

Vitamin D Promotes Ferroptosis in Colorectal Cancer Stem Cells via SLC7A11 Downregulation.

Colorectal cancer stem cells (CCSCs) play important roles in the prognosis, chemoresistance, and treatment failure of colorectal cancer (CRC). Ferroptosis is an effective treatment for CCSCs. Vitamin D (VD) reportedly inhibits colon cancer cell proliferation. However, information on the relationship between VD and ferroptosis in CCSCs is not well documented. In this study, we aimed to understand the effect of VD on ferroptosis in CCSCs. To this end, we treated CCSCs with different concentrations of VD and performed spheroid formation assay and transmission electron microscopy and determined cysteine (Cys), glutathione (GSH), and reactive oxygen species (ROS) levels. Furthermore, functional experiments, western blotting, and qRT-PCR were performed to explore the downstream molecular mechanisms of VD in vitro and in vivo. Results showed that VD treatment significantly inhibited the proliferation of CCSCs and reduced the number of tumour spheroids in vitro. Further evaluations showed that the VD-treated CCSCs exhibited significantly higher ROS levels and lower levels of Cys and GSH as well as thickened mitochondrial membranes. Furthermore, the mitochondria in CCSCs were narrowed and ruptured after VD treatment. These results indicated that VD treatment significantly induced ferroptosis in CCSCs. Further exploration showed that SLC7A11 overexpression significantly attenuated VD-induced ferroptosis in vitro and in vivo. Hence, we concluded that VD induces ferroptosis in CCSCs by downregulating SLC7A11 in vitro and in vivo. These results provide new evidence for the therapeutic use of VD in treating CRC and new insights into VD-induced ferroptosis in CCSCs.

Exploring the mechanism of curcumin in the treatment of colon cancer based on network pharmacology and molecular docking.

Objective: Curcumin is a plant polyphenol extracted from the Chinese herb turmeric. It was found that curcumin has good anti-cancer properties in a variety of cancers, but the exact mechanism is not clear. Based on the network pharmacology and molecular docking to deeply investigate the molecular mechanism of curcumin for the treatment of colon cancer, it provides a new research direction for the treatment of colon cancer. Methods: Curcumin-related targets were collected using PharmMapper, SwissTargetPrediction, Targetnet and SuperPred. Colon cancer related targets were obtained using OMIM, DisGeNET, GeneCards and GEO databases. Drug-disease intersection targets were obtained via Venny 2.1.0. GO and KEGG enrichment analysis of drug-disease common targets were performed using DAVID. Construct PPI network graphs of intersecting targets using STRING database as well as Cytoscape 3.9.0 and filter core targets. Molecular docking via AutoDockTools 1.5.7. The core targets were further analyzed by GEPIA, HPA, cBioPortal and TIMER databases. Results: A total of 73 potential targets of curcumin for the treatment of colon cancer were obtained. GO function enrichment analysis yielded 256 entries, including BP(Biological Progress):166, CC(celluar component):36 and MF(Molecular Function):54. The KEGG pathway enrichment analysis yielded 34 signaling pathways, mainly involved in Metabolic pathways, Nucleotide metabolism, Nitrogen metabolism, Drug metabolism - other enzymes, Pathways in cancer,PI3K-Akt signaling pathway, etc. CDK2, HSP90AA1, AURKB, CCNA2, TYMS, CHEK1, AURKA, DNMT1, TOP2A, and TK1 were identified as core targets by Cytoscape 3.9.0. Molecular docking results showed that the binding energies of curcumin to the core targets were all less than 0kJ-mol-1, suggesting that curcumin binds spontaneously to the core targets. These results were further validated in terms of mRNA expression levels, protein expression levels and immune infiltration. Conclusion: Based on network pharmacology and molecular docking initially revealed that curcumin exerts its therapeutic effects on colon cancer with multi-target, multi-pathway. Curcumin may exert anticancer effects by binding to core targets. Curcumin may interfere with colon cancer cell proliferation and apoptosis by regulating signal transduction pathways such as PI3K-Akt signaling pathway,IL-17 signaling pathway, Cell cycle. This will deepen and enrich our understanding of the potential mechanism of curcumin against colon cancer and provide a theoretical basis for subsequent studies.

Litchi procyanidins inhibit colon cancer proliferation and metastasis by triggering gut-lung axis immunotherapy

Litchi chinensis seed, as a valuable by-product of the subtropical fruit litchi (Litchi chinensis Sonn.), has been confirmed to be rich in procyanidins (LPC). The anticarcinogenic properties of procyanidins has been primarily attributed to their antioxidant and anti-inflammatory activities. However, there is a comparative paucity of information on if and how LPC inhibits colon cancer. Here, LPC significantly inhibited CT26 colon cancer cells proliferation and metastasis in vivo and in vitro. In CT26 lung metastatic mice, the anti-metastatic effect of LPC relied on its regulation of gut microbiota such as increase of Lachnospiraceae UCG-006, Ruminococcus, and their metabolites such as acetic acid, propionic acid and butyric acid. In addition, LPC significantly inhibited CT26 colon cancer cells metastasis through increasing CD8+ cytotoxic T lymphocytes infiltration and decreasing the number of macrophages. Antibiotics treatment demonstrated that the therapeutic effect of LPC depended on the gut microbiota, which regulated T cells immune response. Taken together, LPC had strong inhibitory effects on colon cancer pulmonary metastasis by triggering gut-lung axis to influence the T cells immune response. Our research provides a novel finding for the utilization of procyanidins in the future, that is, supplementing more fruits and vegetables rich in procyanidins is beneficial to the treatment of colon cancer, or it can be used as an adjuvant drug in clinical anti-tumor immunotherapy.

Targeting BAP1 with small compound inhibitor for colon cancer treatment

BRCA1-associated protein-1 (BAP1) is a ubiquitin C-terminal hydrolase domain-containing deubiquitinase. The gene encoding BAP1 is mutated in various human cancers, including mesothelioma, uveal melanoma and renal cell carcinoma. BAP1 plays roles in many cancer-related cellular functions, including cell proliferation, cell death, and nuclear processes crucial for genome stability, such as DNA repair and replication. While these findings suggest that BAP1 functions as a tumor suppressor, recent data also suggest that BAP1 might play tumor-promoting roles in certain cancers, such as breast cancer and hematopoietic malignancies. Here, we show that BAP1 is upregulated in colon cancer cells and tissues and that BAP1 depletion reduces colon cancer cell proliferation and tumor growth. BAP1 contributes to colon cancer cell proliferation by accelerating DNA replication and suppressing replication stress and concomitant apoptosis. A recently identified BAP1 inhibitor, TG2-179-1, which seems to covalently bind to the active site of BAP1, exhibits potent cytotoxic activity against colon cancer cells, with half-maximal inhibitory concentrations of less than 10 μM, and inhibits colon tumor growth. TG2-179-1 exerts cytotoxic activity by targeting BAP1, leading to defective replication and increased apoptosis. This work therefore shows that BAP1 acts oncogenically in colon cancer and is a potential therapeutic target for this cancer. Our work also suggests that TG2-179-1 can be developed as a potential therapeutic agent for colon cancer.