Screening of Alternative Drugs to the Tumor Suppressor miR-375 in Esophageal Squamous Cell Carcinoma Using the Connectivity Map

Abstract

Objective: The aim of this study was to identify alternative compounds to the tumor suppressor miR-375 using the connectivity map (CMAP) and to validate the antitumor effects of the identified drugs in esophageal squamous cell carcinoma (ESCC). Methods: Gene profiling of miR-375-treated TE2 and T.Tn cells was applied in order to search the CMAP database. Among the compounds identified using the CMAP, we focused on 8 drugs [(-)-epigallocatechin-3-gallate, metformin, rosiglitazone among others], excluding 2 drugs among the top 10 compounds. We evaluated whether these compounds possess tumor-suppressive functions in ESCC. Results: A cytotoxicity assay showed that the sensitivity of TE2 and T.Tn cells treated with the 8 compounds was evaluated based on IC₅₀ values of 42.9 µ<smlcap>M</smlcap> to 3.8 m<smlcap>M</smlcap>. A cell cycle analysis revealed that the percentage of TE2 and T.Tn cells incubated with 6 compounds in the G0/G1 phase or the G2/M phase increased by approximately 40-80%. A TUNEL assay showed that the percentages of apoptotic cells treated with almost all compounds were significantly increased (p < 0.05) compared with the control cells. Conclusion: The CMAP database is a useful tool for identifying compounds affecting the same molecular pathways, particularly products that are difficult to apply via practical approaches, such as microRNAs.

Antitumor effects of metformin are a result of inhibiting nuclear factor kappa B nuclear translocation in esophageal squamous cell carcinoma

“…We searched drugs which induce similar gene expression style by connectivity map and found metformin. It restrained the increase of ESCC cell lines, TE2 and T.Tn …”

Metformin in Esophageal Carcinoma: Exploring Molecular Mechanisms and Therapeutic Insights

“…Unsupervised clustering analysis illustrated a distinct miRNA expression profile in metformin-treated KYSE30 cells [ 50 ]. Additional studies examined the gene expression profiles of numerous miRNAs in ESCC cell lines [ 67 , 68 , 69 ]. These analyses employed microarray data in conjunction with the Connectivity Map (CMAP) database, revealing crucial biological functions associated with these miRNAs.…”

“…These analyses employed microarray data in conjunction with the Connectivity Map (CMAP) database, revealing crucial biological functions associated with these miRNAs. These functions encompassed processes such as development, differentiation, apoptosis, and proliferation, elucidating the intricate connections between metformin, genes, and the pathogenesis of ESCC [ 67 , 68 , 69 ] In particular, metformin might activate the TMCO3 and PLA2G4A genes, which are tumor suppressor genes upregulated by miR-375 and have a tumor-suppressive function in themselves as an alternative substance to miR-375 [ 67 ]. Investigating the communication between hypoxic and normoxic cells via exosomes, this study identified exo-miR-340-5p as a key player in the transfer of radioresistance.…”

Metformin-Induced Heat Shock Protein Family A Member 6 Is a Promising Biomarker of Esophageal Squamous Cell Carcinoma

“…Metformin inhibits the proliferation of ESCC cells [11]. We previously reported that microRNA-375 exerted an antitumor effect on ESCC cells and that metformin altered gene expression and inhibited ESCC cell proliferation similar to microRNA-375 [12]. We also reported that nuclear factor-kappa B was important for mediating the antitumor effects of metformin, such as inhibition of tumor growth and epithelial-mesenchymal transition [13].…”