RASSF1A-Hippo pathway link in patients with urothelial carcinoma of bladder: plausible therapeutic target.


Journal

Molecular and cellular biochemistry
ISSN: 1573-4919
Titre abrégé: Mol Cell Biochem
Pays: Netherlands
ID NLM: 0364456

Informations de publication

Date de publication:
Jan 2020
Historique:
received: 20 05 2019
accepted: 03 11 2019
pubmed: 23 11 2019
medline: 19 3 2020
entrez: 23 11 2019
Statut: ppublish

Résumé

RASSF1A is a tumor suppressor gene, and its hypermethylation has been observed in cancers. RASSF1A acts as an upstream regulator of Hippo pathway and modulates its function. The aim of this study was to analyze expression of RASSF1A, Hippo pathway molecules (YAP, MST) and downstream targets (CTGF, Cyr61 and AREG) in bladder cancer patients. Later, the link between RASSF1A and Hippo pathway and a potential therapeutic scope of this link in UBC were also studied. MSPCR was performed to study methylation of RASSF1A promoter. Expression of molecules was studied using qPCR, Western blot and IHC. The link between RASSF1A and Hippo pathway was studied using Spearman's correlation in patients and validated by overexpressing RASSF1A in HT1376 cells and its effect on Hippo pathway was observed using qPCR and Western blot. Further therapeutic potential of this link was studied using MTT and PI assays. The expression of RASSF1A was lower, whereas the expression of YAP, CTGF and CYR61 was higher. The expression of RASSF1A protein gradually decreased, while the expression of YAP, CTGF and CYR61 increased with severity of disease. Based on Spearman's correlation, RASSF1A showed a negative correlation with YAP, CTGF and CYR61. YAP showed a positive correlation with CTGF and CYR61. To validate this link, RASSF1A was overexpressed in HT1376 cells. Overexpressed RASSF1A activated Hippo pathway, followed by a decrease in CTGF and CYR61 at mRNA, and enhanced cytotoxicity to chemotherapeutic drugs. This study finds a previously unrecognized role of RASSF1A in the regulation of CTGF and CYR61 through mediation of Hippo pathway in UBC and supports the significance of this link as a potential therapeutic target for UBC.

Identifiants

pubmed: 31754973
doi: 10.1007/s11010-019-03648-y
pii: 10.1007/s11010-019-03648-y
doi:

Substances chimiques

CCN1 protein, human 0
CCN2 protein, human 0
Cysteine-Rich Protein 61 0
RASSF1 protein, human 0
Tumor Suppressor Proteins 0
Connective Tissue Growth Factor 139568-91-5
Protein Serine-Threonine Kinases EC 2.7.11.1

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

51-63

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Auteurs

Madhuram Khandelwal (M)

Department of Biochemistry, AIIMS, New Delhi, India.

Vivek Anand (V)

Department of Biochemistry, AIIMS, New Delhi, India.

Sandeep Appunni (S)

Department of Biochemistry, AIIMS, New Delhi, India.

Amlesh Seth (A)

Department of Urology, AIIMS, New Delhi, India.

Prabhjot Singh (P)

Department of Urology, AIIMS, New Delhi, India.

Sandeep Mathur (S)

Department of Pathology, AIIMS, New Delhi, India.

Alpana Sharma (A)

Department of Biochemistry, AIIMS, New Delhi, India. dralpanasharma@aiims.edu.

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