A kinome-centered CRISPR-Cas9 screen identifies activated BRAF to modulate enzalutamide resistance with potential therapeutic implications in BRAF-mutated prostate cancer

AbstractResistance to drugs targeting the androgen receptor (AR) signaling axis remains an important challenge in the treatment of prostate cancer patients. Activation of alternative growth pathways is one mechanism used by cancer cells to proliferate despite treatment, conferring drug resistance. Through a kinome-centered CRISPR-Cas9 screen in CWR-R1 prostate cancer cells, we identified activated BRAF signaling as a determinant for enzalutamide resistance. Combined pharmaceutical targeting of AR and MAPK signaling resulted in strong synergistic inhibition of cell proliferation. The association between BRAF activation and enzalutamide resistance was confirmed in two metastatic prostate cancer patients harboring activating mutations in the BRAF gene, as both patients were unresponsive to enzalutamide. Our findings suggest that co-targeting of the MAPK and AR pathways may be effective in patients with an activated MAPK pathway, particularly in patients harboring oncogenic BRAF mutations. These results warrant further investigation of the response to AR inhibitors in BRAF-mutated prostate tumors in clinical settings.

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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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Long non-coding RNA H19 acts as a microRNA-194 sponge to inhibit the apoptosis and promote the proliferation of hypertrophic scar fibroblasts

Canadian Journal of Physiology and Pharmacology ◽

10.1139/cjpp-2021-0351 ◽

2021 ◽

Author(s):

Bo Liu ◽

Lijuan Lin ◽

Shengjin Yu ◽

Rongjun Xia ◽

Linlin Zheng

Keyword(s):

P38 Mapk ◽

Hypertrophic Scar ◽

Mapk Pathway ◽

Mapk Signaling ◽

Hypertrophic Scars ◽

Downstream Effector ◽

Non Coding Rna ◽

Proliferation And Apoptosis ◽

Signaling Axis ◽

Underlying Mechanisms

The effects of long non-coding RNAs (lncRNAs) on the proliferation of hypertrophic scars have been described. However, the underlying mechanisms are not well characterized. The present study aimed to investigate the mechanisms of lncRNA H19 in hypertrophic scars. The effects of the lncRNA H19 on the proliferation and apoptosis of hypertrophic scar fibroblasts (HSFs) were analyzed using 5’-Ethynyl-2’-deoxyuridine staining, flow cytometry, and MTT. The results revealed H19 promoted the proliferation and inhibited the apoptosis in HSF. In addition, the binding associations between H19 and microRNA-194 (miR-194), and miR-194 and insulin-like growth factor-I receptor (IGF1R) were identified using bioinformatics screening and verified using dual-luciferase assays. Furthermore, the effects of the IGF1R knockdown on H19-induced HSF phenotypes and regulation over the p38 MAPK pathway were determined. Mechanistically, miR-194 was identified as the downstream effector of the H19-mediated phenotypes of HSFs through its ability to directly target IGF1R, thus modulating the p38 MAPK signaling pathway. In conclusion, the findings suggested that H19 may inhibit the apoptosis and promote the proliferation of HSFs through the miR-194/IGF1R/p38 MAPK signaling axis, thereby contributing to the progression of hypertrophic scars. These findings may provide novel targets for the treatment of hypertrophic scars.

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Nimbolide Inhibits Cell Survival and Proliferation “ of IGF1 Mediated PI3K-Akt and MAPK Signaling in Human Breast Cancer Cells and TNF-A/TNFR1 Mediated Signaling Molecules in Prostate Cancer

Modern Applications of Bioequivalence & Bioavailability ◽

10.19080/mabb.2017.01.555554 ◽

2017 ◽

Vol 1 (1) ◽

Author(s):

J Arunakaran

Keyword(s):

Breast Cancer ◽

Prostate Cancer ◽

Cell Survival ◽

Cancer Cells ◽

Human Breast Cancer ◽

Breast Cancer Cells ◽

Mapk Signaling ◽

Signaling Molecules ◽

Human Breast Cancer Cells ◽

Human Breast

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Cellular and Molecular Mechanisms Underlying Prostate Cancer Development: Therapeutic Implications

Medicines ◽

10.3390/medicines6030082 ◽

2019 ◽

Vol 6 (3) ◽

pp. 82 ◽

Cited By ~ 12

Author(s):

Ugo Testa ◽

Germana Castelli ◽

Elvira Pelosi

Keyword(s):

Prostate Cancer ◽

Molecular Mechanisms ◽

Intraepithelial Neoplasia ◽

Cancer Development ◽

Initial Development ◽

Therapeutic Implications ◽

Cancer Pathways ◽

Signaling Axis ◽

Prostate Cancer Development ◽

Prostate Cancers

Prostate cancer is the most frequent nonskin cancer and second most common cause of cancer-related deaths in man. Prostate cancer is a clinically heterogeneous disease with many patients exhibiting an aggressive disease with progression, metastasis, and other patients showing an indolent disease with low tendency to progression. Three stages of development of human prostate tumors have been identified: intraepithelial neoplasia, adenocarcinoma androgen-dependent, and adenocarcinoma androgen-independent or castration-resistant. Advances in molecular technologies have provided a very rapid progress in our understanding of the genomic events responsible for the initial development and progression of prostate cancer. These studies have shown that prostate cancer genome displays a relatively low mutation rate compared with other cancers and few chromosomal loss or gains. The ensemble of these molecular studies has led to suggest the existence of two main molecular groups of prostate cancers: one characterized by the presence of ERG rearrangements (~50% of prostate cancers harbor recurrent gene fusions involving ETS transcription factors, fusing the 5′ untranslated region of the androgen-regulated gene TMPRSS2 to nearly the coding sequence of the ETS family transcription factor ERG) and features of chemoplexy (complex gene rearrangements developing from a coordinated and simultaneous molecular event), and a second one characterized by the absence of ERG rearrangements and by the frequent mutations in the E3 ubiquitin ligase adapter SPOP and/or deletion of CDH1, a chromatin remodeling factor, and interchromosomal rearrangements and SPOP mutations are early events during prostate cancer development. During disease progression, genomic and epigenomic abnormalities accrued and converged on prostate cancer pathways, leading to a highly heterogeneous transcriptomic landscape, characterized by a hyperactive androgen receptor signaling axis.

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Diallyl Trisulfide Is More Cytotoxic to Prostate Cancer Cells PC-3 than to Noncancerous Epithelial Cell Line PNT1A: A Possible Role of p66Shc signaling Axis

Nutrition and Cancer ◽

10.1080/01635581.2013.789115 ◽

2013 ◽

Vol 65 (5) ◽

pp. 711-717 ◽

Cited By ~ 15

Author(s):

Andzelika Borkowska ◽

Narcyz Knap ◽

Jędrzej Antosiewicz

Keyword(s):

Prostate Cancer ◽

Epithelial Cell ◽

Cancer Cells ◽

Cell Line ◽

Epithelial Cell Line ◽

Diallyl Trisulfide ◽

Prostate Cancer Cells ◽

Signaling Axis

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Naringenin-Induced Apoptotic Cell Death in Prostate Cancer Cells Is Mediated via the PI3K/AKT and MAPK Signaling Pathways

Journal of Cellular Biochemistry ◽

10.1002/jcb.25729 ◽

2017 ◽

Vol 118 (5) ◽

pp. 1118-1131 ◽

Cited By ~ 44

Author(s):

Whasun Lim ◽

Sunwoo Park ◽

Fuller W. Bazer ◽

Gwonhwa Song

Keyword(s):

Prostate Cancer ◽

Cell Death ◽

Cancer Cells ◽

Signaling Pathways ◽

Apoptotic Cell ◽

Mapk Signaling ◽

Apoptotic Cell Death ◽

Prostate Cancer Cells ◽

Mapk Signaling Pathways

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Prostate Osteoblast-Like Cells: A Reliable Prognostic Marker of Bone Metastasis in Prostate Cancer Patients

Contrast Media & Molecular Imaging ◽

10.1155/2018/9840962 ◽

2018 ◽

Vol 2018 ◽

pp. 1-12 ◽

Cited By ~ 6

Author(s):

Manuel Scimeca ◽

Nicoletta Urbano ◽

Bonfiglio Rita ◽

Sarah Natalia Mapelli ◽

Carlo Vittorio Catapano ◽

...

Keyword(s):

Prostate Cancer ◽

Bone Metastasis ◽

Cancer Cells ◽

Cancer Patients ◽

Clinical Evidence ◽

Prostate Cancer Cells ◽

Ultrastructural Studies ◽

Pet Ct ◽

Ct Analysis ◽

Malignant Lesions

The main aim of this study was to investigate the putative association among the presence of prostate cancer cells, defined as prostate osteoblast-like cells (POLCs), and showing the expression of typical morphological and molecular characteristics of osteoblasts, the development of bone metastasis within 5 years of diagnosis, and the uptake of 18F-choline evaluated by PET/CT analysis. To this end, prostate biopsies (n= 110) were collected comprising 44 benign lesions and 66 malignant lesions. Malignant lesions were further subdivided into two groups: biopsies from patients that had clinical evidence of bone metastasis (BM+,n= 23) and biopsies from patients that did not have clinical evidence of bone metastasis within 5 years (BM−,n= 43). Paraffin serial sections were obtained from each specimen to perform histological classifications and immunohistochemical (IHC) analysis. Small fragments of tissue were used to perform ultrastructural and microanalytical investigations. IHC demonstrated the expression of markers of epithelial-to-mesenchymal transition (VIM), bone mineralization, and osteoblastic differentiation (BMP-2, PTX-3, RUNX2, RANKL, and VDR) in prostate lesions characterized by the presence of calcium-phosphate microcalcifications and high metastatic potential. Ultrastructural studies revealed the presence of prostate cancer cells with osteoblast phenotype close to microcalcifications. Noteworthy, PET/CT analysis showed higher uptake of 18F-choline in BM+ lesions with high positivity (≥300/500 cells) for RUNX2 and/or RANKL immunostaining. Although these data require further investigations about the molecular mechanisms of POLCs generation and role in bone metastasis, our study can open new and interesting prospective in the management of prostate cancer patients. The presence of POLCs along with prostate microcalcifications may become negative prognostic markers of the occurrence of bone metastases.

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Glycidamide Promotes the Growth and Migratory Ability of Prostate Cancer Cells by Changing the Protein Expression of Cell Cycle Regulators and Epithelial-to-Mesenchymal Transition (EMT)-Associated Proteins with Prognostic Relevance

International Journal of Molecular Sciences ◽

10.3390/ijms20092199 ◽

2019 ◽

Vol 20 (9) ◽

pp. 2199

Author(s):

Titus Ime Ekanem ◽

Chi-Chen Huang ◽

Ming-Heng Wu ◽

Ding-Yen Lin ◽

Wen-Fu T. Lai ◽

...

Keyword(s):

Prostate Cancer ◽

Cell Cycle ◽

Protein Expression ◽

Cancer Cells ◽

Cancer Patients ◽

Epithelial To Mesenchymal Transition ◽

Gene Signature ◽

Prostate Cancer Cells ◽

Mesenchymal Transition ◽

Prognostic Relevance

Acrylamide (AA) and glycidamide (GA) can be produced in carbohydrate-rich food when heated at a high temperature, which can induce a malignant transformation. It has been demonstrated that GA is more mutagenic than AA. It has been shown that the proliferation rate of some cancer cells are increased by treatment with GA; however, the exact genes that are induced by GA in most cancer cells are not clear. In the present study, we demonstrated that GA promotes the growth of prostate cancer cells through induced protein expression of the cell cycle regulator. In addition, we also found that GA promoted the migratory ability of prostate cancer cells through induced epithelial-to-mesenchymal transition (EMT)-associated protein expression. In order to understand the potential prognostic relevance of GA-mediated regulators of the cell cycle and EMT, we present a three-gene signature to evaluate the prognosis of prostate cancer patients. Further investigations suggested that the three-gene signature (CDK4, TWIST1 and SNAI2) predicted the chances of survival better than any of the three genes alone for the first time. In conclusion, we suggested that the three-gene signature model can act as marker of GA exposure. Hence, this multi-gene panel may serve as a promising outcome predictor and potential therapeutic target in prostate cancer patients.

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