Acrolein contributes to human colorectal tumorigenesis through the activation of RAS-MAPK pathway

AbstractColorectal cancer (CRC) is one of the most well-known malignancies with high prevalence and poor 5-year survival. Previous studies have demonstrated that a high-fat diet (HFD) is capable of increasing the odds of developing CRC. Acrolein, an IARC group 2A carcinogen, can be formed from carbohydrates, vegetable oils, animal fats, and amino acids through the Maillard reaction during the preparation of foods. Consequently, humans are at risk of acrolein exposure through the consumption of foods rich in fat. However, whether acrolein contributes to HFD-induced CRC has not been determined. In this study, we found that acrolein induced oncogenic transformation, including faster cell cycling, proliferation, soft agar formation, sphere formation and cell migration, in NIH/3T3 cells. Using xenograft tumorigenicity assays, the acrolein-transformed NIH/3T3 clone formed tumors. In addition, cDNA microarray and bioinformatics studies by Ingenuity Pathway Analysis pointed to the fact that RAS/MAPK pathway was activated in acrolein-transformed clones that contributed to colon tumorigenesis. Furthermore, acrolein-induced DNA damages (Acr-dG adducts) were higher in CRC tumor tissues than in normal epithelial cells in CRC patients. Notably, CRC patients with higher levels of Acr-dG adducts appeared to have better prognosis. The results of this study demonstrate for the first time that acrolein is important in oncogenic transformation through activation of the RAS/MAPK signaling pathway, contributing to colon tumorigenesis.

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Acrolein Contributes to Human Colorectal Tumorigenesis Through the Activation of RAS/MAPK Pathway.

10.21203/rs.3.rs-142335/v1 ◽

2021 ◽

Author(s):

Hong-Chieh Tsai ◽

Han-Hsing Tsou ◽

Chun-Chi Lin ◽

Shao-Chen Chen ◽

Hsiao-Wei Cheng ◽

...

Keyword(s):

Mapk Pathway ◽

Soft Agar ◽

Mapk Signaling ◽

Glycerol Metabolism ◽

Oncogenic Transformation ◽

Dna Damages ◽

Colorectal Tumorigenesis ◽

Colon Tumorigenesis ◽

Nih 3T3 ◽

Sphere Formation

Abstract Colorectal cancer (CRC) is one of the most well-known malignancies with high prevalence and poor 5-year survival. Previous studies have demonstrated that high-fat diet (HFD) is capable of increasing the odds of developing CRC. Acrolein, an IARC group 2A carcinogen, can be formed through Maillard reaction. Also, acrolein has been shown to be produced from microbial glycerol metabolism in human gut. Consequently, humans are at risk of acrolein exposure through consumption of foods rich in fat. However, whether acrolein contributes to HFD-induced CRC remains elusive. In this study, we found that acrolein induced oncogenic transformation including faster cell cycling, proliferation, soft agar formation, sphere formation and cell migration in NIH/3T3 cells. Using xenograft tumorigenicity assays, the acrolein-transformed NIH/3T3 clone formed tumors. In addition, RAS/MAPK pathway contributing to colon tumorigenesis was activated in acrolein-transformed clones using cDNA microarray analysis with Ingenuity Pathway Analysis. Besides, acrolein-induced DNA damages (Acr-dG adducts) were higher in CRC tumor tissues compared to normal epithelial cells in CRC patients. Intriguingly, CRC patients with higher Acr-dG adducts have better prognosis. Taken together, this is the first study to demonstrate that acrolein is important in oncogenic transformation through activating RAS/MAPK signaling pathway contributing to colon tumorigenesis.

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BTB and CNC homology 1 (Bach1) induces lung cancer stem cell phenotypes by inducing CD44 expression

10.21203/rs.3.rs-122503/v1 ◽

2020 ◽

Author(s):

Pan Jiang ◽

Shengyu Hao ◽

Liang Xie ◽

Guiling Xiang ◽

Weiping Hu ◽

...

Keyword(s):

Lung Cancer ◽

Flow Cytometry ◽

Cancer Metastasis ◽

Mapk Pathway ◽

Mapk Signaling ◽

Luciferase Reporter ◽

Ki 67 ◽

Tumor Sphere ◽

Sphere Formation

Abstract Background Growing evidence suggests that CSCs are responsible for cancer initiation in tumors. Bach1 has been identified to contribute to several tumor progression, including lung cancer. The role of Bach1 in CSCs remains poorly known. Therefore, the function of Bach1 on lung CSCs was focused currently. Methods The expression of Bach1, CD133, CD44, Sox2, Nanog and Oct4 mRNA was assessed using RT-qPCR. Protein expression of Bach1, CD133, CD44, Sox2, Nanog and Oct4 was analyzed by western blotting. EdU, colony formation, Flow cytometry analysis and transwell invasion assay were carried out to analyze cell proliferation, apoptosis and invasion. Tumor sphere formation assay was utilized to evaluate spheroid capacity. The relationship between Bach1 and CD44 was verified using ChIP-qPCR and dual-luciferase reporter assay. Ki-67 expression was determined by flow cytometry and IHC assay. Results The ratio of CD44 + CSCs from A549 and SPC-A1 cells were significantly enriched. Tumor growth of CD44 + CSCs was obviously suppressed in vivo. Bach1 expression was obviously increased in CD44 + CSCs. Then, via using the in vitro experiment, it was observed that CSC growth and invasion were greatly reduced by the down-regulation of Bach1. Loss of Bach1 was able to repress tumor-sphere formation and tumor-initiating CSC markers via acting as a transcription inducer of CD44. A repression of CSCs growth and metastasis of shRNA-Bach1 was confirmed using xenograft models. Furthermore, MAPK signaling pathway was selected and we proved the effects of Bach1 on lung CSCs were associated with the activation of the MAPK pathway. Inhibition of MAPK signaling remarkably restrained lung CSCs growth and CSCs properties. Conclusion In summary, our findings reveal that Bach1 positively regulated CD44 + lung cancer CSCs growth and we imply that Bach1 demonstrates great potential for the treatment of lung cancer metastasis and recurrence via regulating CD44 and MPAK signaling.

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Resistance to oncogenic transformation in revertant R1 of human ras-transformed NIH 3T3 cells.

Molecular and Cellular Biology ◽

10.1128/mcb.9.5.2258 ◽

1989 ◽

Vol 9 (5) ◽

pp. 2258-2263 ◽

Cited By ~ 17

Author(s):

N Kuzumaki ◽

Y Ogiso ◽

A Oda ◽

H Fujita ◽

H Suzuki ◽

...

Keyword(s):

Simian Virus 40 ◽

Soft Agar ◽

Simian Virus ◽

T Antigen ◽

Oncogenic Transformation ◽

3T3 Cells ◽

Cellular Mechanisms ◽

Ras Gene ◽

Nih 3T3 ◽

Nih 3T3 Cells

A flat revertant, R1, was isolated from human activated c-Ha-ras-1 (hu-ac-Ha-ras) gene-transformed NIH 3T3 cells (EJ-NIH 3T3) treated with mutagens. R1 contained unchanged transfected hu-ac-Ha-ras DNA and expressed high levels of hu-ac-Ha-ras-specific mRNA and p21 protein. Transfection experiments revealed that NIH 3T3 cells could be transformed by DNA from R1 cells but R1 cells could not be retransformed by Kirsten sarcoma virus, DNA from EJ-NIH 3T3 cells, hu-ac-Ha-ras, v-src, v-mos, simian virus 40 large T antigen, or polyomavirus middle T antigen. Somatic cell hybridization studies showed that R1 was not retransformed by fusion with NIH 3T3 cells and suppressed anchorage independence of EJ-NIH 3T3 and hu-ac-Ha-ras gene-transformed rat W31 cells in soft agar. These results suggest that the reversion and resistance to several oncogenes in R1 is due not to cellular defects in the production of the transformed phenotype but rather to enhancement of cellular mechanisms that suppress oncogenic transformation.

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Acrolein Contributes to Human Colorectal Tumorigenesis Through Activation of RAS/MAPK Pathway

10.21203/rs.3.rs-139690/v1 ◽

2021 ◽

Author(s):

Hong-Chieh Tsai ◽

Han-Hsing Tsou ◽

Chun-Chi Lin ◽

Shao-Chen Chen ◽

Hsiao-Wei Cheng ◽

...

Keyword(s):

Microarray Analysis ◽

Cdna Microarray ◽

Oncogenic Transformation ◽

3T3 Cells ◽

Cdna Microarray Analysis ◽

Colon Tumorigenesis ◽

Tumor Tissues ◽

Chi Square Analysis ◽

Nih 3T3 ◽

Nih 3T3 Cells

Abstract Background. Colorectal cancer (CRC) is one of the most well-known malignancies with high prevalence and poor 5-year survival. Previous studies have demonstrated that intake of food rich in fat, and with low fiber content (as known as high-fat diet, HFD) is capable of increasing the odds of developing CRC. Acrolein, an IARC group 2A carcinogen, can be formed by thermal treatment of animal and vegetable fats, carbohydrates and amino acids through Maillard reaction. Also, acrolein has been shown to be produced from microbial glycerol metabolism in human gut. Consequently, humans are at risk of acrolein exposure through consumption of foods rich in fat. However, whether acrolein contributes to HFD-induced CRC tumorigenesis remains elusive.Methods. The effect of acrolein in oncogenic transformation was analyzed using NIH/3T3 cells with xenograft tumorigenesis mice models. Furthermore, cDNA microarray analysis with Ingenuity Pathway Analysis (IPA) was performed in acrolein-transformed NIH/3T3 cells. Finally, acrolein-induced DNA damages (Acr-dG) were analyzed in tumor tissues and normal epithelial of CRC patients using immunohistochemical analysis. The levels of Acr-dG adducts were associated with tumor characteristics and CRC patients’ survival using Chi-Square analysis and Kaplan Meier survival analysis, respectively.Results. In this present study, we found that acrolein induced oncogenic transformation including faster cell cycling, proliferation, soft agar formation, sphere formation, cell migration in NIH/3T3 cells. Using xenograft tumorigenicity assays, the acrolein-transformed NIH3T3 clone formed tumors whereas no tumors were observed in mice inoculated with NIH3T3 parental cells. In addition, RAS/MAPK pathway contributing to colon tumorigenesis was activated in NIH/3T3 Acr-clone using cDNA microarray analysis with IPA. Finally, Acr-dG adducts were higher in CRC tumor tissues compared to normal epithelial cells in CRC patients. Intriguingly, CRC patients with higher Acr-dG adducts have better prognosis. Conclusions. Taken together, this is the first study to demonstrate that acrolein is important in oncogenic transformation through activating RAS/MAPK signaling pathway contributing to colon tumorigenesis. Thus, acrolein might be a novel target for early detection, prevention and treatment of tumors in the future.

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Resistance to oncogenic transformation in revertant R1 of human ras-transformed NIH 3T3 cells

Molecular and Cellular Biology ◽

10.1128/mcb.9.5.2258-2263.1989 ◽

1989 ◽

Vol 9 (5) ◽

pp. 2258-2263

Author(s):

N Kuzumaki ◽

Y Ogiso ◽

A Oda ◽

H Fujita ◽

H Suzuki ◽

...

Keyword(s):

Simian Virus 40 ◽

Soft Agar ◽

Simian Virus ◽

T Antigen ◽

Oncogenic Transformation ◽

3T3 Cells ◽

Cellular Mechanisms ◽

Ras Gene ◽

Nih 3T3 ◽

Nih 3T3 Cells

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Brassinin Inhibits Proliferation in Human Liver Cancer Cells via Mitochondrial Dysfunction

Cells ◽

10.3390/cells10020332 ◽

2021 ◽

Vol 10 (2) ◽

pp. 332

Author(s):

Taeyeon Hong ◽

Jiyeon Ham ◽

Jisoo Song ◽

Gwonhwa Song ◽

Whasun Lim

Keyword(s):

Mitochondrial Dysfunction ◽

Cancer Cells ◽

Cell Lines ◽

Mapk Pathway ◽

Mapk Signaling ◽

Nuclear Antigen ◽

Cruciferous Vegetable ◽

Antiproliferative Effects ◽

Hep3b Cells ◽

Human Liver Cancer Cells

Brassinin is a phytochemical derived from Chinese cabbage, a cruciferous vegetable. Brassinin has shown anticancer effects on prostate and colon cancer cells, among others. However, its mechanisms and effects on hepatocellular carcinoma (HCC) have not been elucidated yet. Our results confirmed that brassinin exerted antiproliferative effects by reducing proliferating cell nuclear antigen (PCNA) activity, a proliferation indicator and inducing cell cycle arrest in human HCC (Huh7 and Hep3B) cells. Brassinin also increased mitochondrial Ca2+ levels and depolarized the mitochondrial membrane in both Huh7 and Hep3B cells. Moreover, brassinin generated high amounts of reactive oxygen species (ROS) in both cell lines. The ROS scavenger N-acetyl-L-cysteine (NAC) inhibited this brassinin-induced ROS production. Brassinin also regulated the AKT and mitogen-activated protein kinases (MAPK) signaling pathways in Huh7 and Hep3B cells. Furthermore, co-administering brassinin and pharmacological inhibitors for JNK, ERK1/2 and P38 decreased cell proliferation in both HCC cell lines more than the pharmacological inhibitors alone. Collectively, our results demonstrated that brassinin exerts antiproliferative effects via mitochondrial dysfunction and MAPK pathway regulation on HCC cells.

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Antimetastatic Effects of Sesamin on Human Head and Neck Squamous Cell Carcinoma through Regulation of Matrix Metalloproteinase-2

Molecules ◽

10.3390/molecules25092248 ◽

2020 ◽

Vol 25 (9) ◽

pp. 2248 ◽

Cited By ~ 1

Author(s):

Jian-Ming Chen ◽

Pei-Yin Chen ◽

Chia-Chieh Lin ◽

Ming-Chang Hsieh ◽

Jen-Tsun Lin

Keyword(s):

Oral Cancer ◽

Matrix Metalloproteinase ◽

Head And Neck ◽

Mapk Pathway ◽

Human Head ◽

Mapk Signaling ◽

Matrix Metalloproteinase 2 ◽

Migration And Invasion ◽

Dependent Manner

Background: Sesamin is a lignin present in sesame oil from the bark of Zanthoxylum spp. Sesamin reportedly has anticarcinogenic potential and exerts anti-inflammatory effects on several tumors. Hypothesis/Purpose: However, the effect of sesamin on metastatic progression in human head and neck squamous carcinoma (HNSCC) remains unknown in vitro and in vivo; hence, we investigated the effect of sesamin on HNSCC cells in vitro. Methods and Results: Sesamin-treated human oral cancer cell lines FaDu, HSC-3, and Ca9-22 were subjected to a wound-healing assay. Furthermore, Western blotting was performed to assess the effect of sesamin on the expression levels of matrix metalloproteinase (MMP)-2 and proteins of the MAPK signaling pathway, including p-ERK1/2, P-p38, and p-JNK1/2. In addition, we investigated the association between MMP-2 expression and the MAPK pathway in sesamin-treated oral cancer cells. Sesamin inhibited cell migration and invasion in FaDu, Ca9-22, and HSC-3 cells and suppressed MMP-2 at noncytotoxic concentrations (0 to 40 μM). Furthermore, sesamin significantly reduced p38 MAPK and JNK phosphorylation in a dose-dependent manner in FaDu and HSC-3 cells. Conclusions: These results indicate that sesamin suppresses the migration and invasion of HNSCC cells by regulating MMP-2 and is thus a potential antimetastatic agent for treating HNSCC.

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Epigenetic control of pheromone MAPK signaling determines sexual fecundity inCandida albicans

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1711141115 ◽

2017 ◽

Vol 114 (52) ◽

pp. 13780-13785 ◽

Cited By ~ 6

Author(s):

Christine M. Scaduto ◽

Shail Kabrawala ◽

Gregory J. Thomson ◽

William Scheving ◽

Andy Ly ◽

...

Keyword(s):

Candida Albicans ◽

G Protein ◽

Map Kinases ◽

Mapk Pathway ◽

Cell Types ◽

Mapk Signaling ◽

Toggle Switch ◽

Β Subunit ◽

Epigenetic Control ◽

Incandida Albicans

Several pathogenicCandidaspecies are capable of heritable and reversible switching between two epigenetic states, “white” and “opaque.” InCandida albicans, white cells are essentially sterile, whereas opaque cells are mating-proficient. Here, we interrogate the mechanism by which the white-opaque switch regulates sexual fecundity and identify four genes in the pheromone MAPK pathway that are expressed at significantly higher levels in opaque cells than in white cells. These genes encode the β subunit of the G-protein complex (STE4), the pheromone MAPK scaffold (CST5), and the two terminal MAP kinases (CEK1/CEK2). To define the contribution of each factor to mating,C. albicanswhite cells were reverse-engineered to express elevated, opaque-like levels of these factors, either singly or in combination. We show that white cells co-overexpressingSTE4,CST5, andCEK2undergo mating four orders of magnitude more efficiently than control white cells and at a frequency approaching that of opaque cells. Moreover, engineered white cells recapitulate the transcriptional and morphological responses of opaque cells to pheromone. These results therefore reveal multiple bottlenecks in pheromone MAPK signaling in white cells and that alleviation of these bottlenecks enables efficient mating by these “sterile” cell types. Taken together, our findings establish that differential expression of several MAPK factors underlies the epigenetic control of mating inC. albicans. We also discuss how fitness advantages could have driven the evolution of a toggle switch to regulate sexual reproduction in pathogenicCandidaspecies.

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The K-fgf/hst oncogene induces transformation through an autocrine mechanism that requires extracellular stimulation of the mitogenic pathway

Molecular and Cellular Biology ◽

10.1128/mcb.11.2.1138-1145.1991 ◽

1991 ◽

Vol 11 (2) ◽

pp. 1138-1145

Author(s):

D Talarico ◽

C Basilico

Keyword(s):

Growth Factor ◽

Receptor Activation ◽

Soft Agar ◽

Serum Free Medium ◽

Free Medium ◽

3T3 Cells ◽

Serum Free ◽

Nih 3T3 ◽

Autocrine Mechanism ◽

Nih 3T3 Cells

The K-fgf/hst oncogene encodes a secreted growth factor of the fibroblast growth factor (FGF) family. The ability of K-fgf-transformed cells to grow in soft agar and in serum-free medium is inhibited by anti-K-FGF neutralizing antibodies, consistent with an autocrine mechanism of transformation. The transformed properties of clones that express high levels of K-FGF are, however, only partially affected. To better define the autocrine mechanism of transformation by K-fgf and to determine whether receptor activation could occur intracellularly, we constructed two mutants of the K-fgf cDNA. Deletion of the sequences encoding the signal peptide suppressed K-fgf ability to induce foci in NIH 3T3 cells. A few morphologically transformed colonies were observed in cotransfection experiments, and they were found to express high levels of cytoplasmic K-FGF. However, their ability to grow in serum-free medium and in soft agar was inhibited by anti-K-FGF antibodies. Addition of a sequence encoding the KDEL endoplasmic reticulum and Golgi retention signal to the K-fgf cDNA led to accumulation of the growth factor in intracellular compartments. The ability of the KDEL mutant to induce foci in NIH 3T3 cells was much lower than that of the wild-type cDNA, and also in this case the transformed phenotype was reverted by anti-K-FGF antibodies. These and other findings indicate that the transformed phenotype of cells expressing a nonsecretory K-FGF is due to the extracellular activation of the receptor by the small amounts of growth factor that these cells still release. Thus, transformation by K-fgf appears to be due to an autocrine growth mechanisms that requires activation of the mitogenic pathway at the cell surface.

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The defense response of Caenorhabditis elegans to Cutibacterium acnes SK137 via the TIR-1-p38 MAPK signaling pathway

Bioscience Biotechnology and Biochemistry ◽

10.1093/bbb/zbab218 ◽

2021 ◽

Author(s):

Ayano Tsuru ◽

Yumi Hamazaki ◽

Shuta Tomida ◽

Mohammad Shaokat Ali ◽

Eriko Kage-Nakadai

Keyword(s):

Caenorhabditis Elegans ◽

Signaling Pathway ◽

P38 Mapk ◽

Defense Response ◽

Mapk Pathway ◽

Defense Responses ◽

Mapk Signaling ◽

Dependent Manner ◽

Healthy Skin ◽

Cutibacterium Acnes

Abstract Cutibacterium acnes plays roles in both acne disease and healthy skin ecosystem. We observed that mutations in the tir-1/SARM1 and p38 MAPK cascade genes significantly shortened Caenorhabditis elegans lifespan upon Cutibacterium acnes SK137 infection. Antimicrobial molecules were induced by SK137 in a TIR-1-dependent manner. These results suggest that defense responses against SK137 involve the TIR-1-p38 MAPK pathway in Caenorhabditis elegans.

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