ABSTRACT
BACKGROUND: Hematoporphyrin derivative (HPD) has a sensibilization effect in lung adenocarcinoma. This study was conducted to identify the target genes of HPD in lung adenocarcinoma. METHODS: RNA sequencing was performed using the lung adenocarcinoma cell line A549 after no treatment or treatment with X-ray or X-ray + HPD. The differentially expressed genes (DEGs) were screened using Mfuzz package by noise-robust soft clustering analysis. Enrichment analysis was carried out using "BioCloud" online tool. Protein-protein interaction (PPI) network and module analyses were performed using Cytoscape software. Using WebGestalt tool and integrated transcription factor platform (ITFP), microRNA target and transcription factor (TF) target pairs were separately predicted. An integrated regulatory network was visualized with Cytoscape software. RESULTS: A total of 815 DEGs in the gene set G1 (continuously dysregulated genes along with changes in processing conditions [untreated-treated with X-ray-X-ray + treated with HPD]) and 464 DEGs in the gene set G2 (significantly dysregulated between X-ray + HPD-treated group and untreated/X-ray-treated group) were screened. The significant module identified from the PPI network for gene set G1 showed that ribosomal protein L3 (RPL3) gene could interact with heat shock protein 90 kDa alpha, class A member 1 (HSP90AA1). TFs AAA domain containing 2 (ATAD2) and protein inhibitor of activated STAT 1 (PIAS1) were separately predicted for the genes in gene set G1 and G2, respectively. In the integrated network for gene set G2, ubiquitin-specific peptidase 25 (USP25) was targeted by miR-200b, miR-200c, and miR-429. CONCLUSION: RPL3, HSP90AA1, ATAD2, and PIAS1 as well as USP25, which is targeted by miR-200b, miR-200c, and miR-429, may be the potential targets of HPD in lung adenocarcinoma.
Subject(s)
Humans , Hematoporphyrin Derivative/pharmacology , Gene Regulatory Networks/genetics , Adenocarcinoma of Lung/genetics , Lung Neoplasms/genetics , Ribosomal Proteins/drug effects , Ribosomal Proteins/genetics , Transcription Factors , Cluster Analysis , Gene Expression Regulation, Neoplastic , Sequence Analysis, RNA , HSP90 Heat-Shock Proteins/drug effects , HSP90 Heat-Shock Proteins/genetics , Small Ubiquitin-Related Modifier Proteins/drug effects , Small Ubiquitin-Related Modifier Proteins/genetics , MicroRNAs/metabolism , Cell Line, Tumor , DNA-Binding Proteins/drug effects , DNA-Binding Proteins/genetics , Protein Inhibitors of Activated STAT/drug effects , Protein Inhibitors of Activated STAT/genetics , Flow Cytometry , ATPases Associated with Diverse Cellular Activities/drug effects , ATPases Associated with Diverse Cellular Activities/genetics , Adenocarcinoma of Lung/drug therapy , Adenocarcinoma of Lung/radiotherapy , Lung Neoplasms/drug therapy , Lung Neoplasms/radiotherapyABSTRACT
Effect of hematoporphyrin derivative (HpD) and light on sulfhydryl (SH) groups in brain tumor cells was studied. Sulfhydryl groups were measured by 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB) and a fluorescent probe 3,4-maleimidylphenyl-4-methyl-7-diethylaminocoumarin (CPM). Incubation of cells with HpD in dark resulted in the loss of DTNB as well as CPM reactive SH groups. After 2 hr of incubation DTNB reactive SH groups showed a negligible change while a continuous decrease was observed in CPM reactive SH groups. Cells treated with HpD showed a further degradation of SH groups upon light irradiation. A comparison of cytotoxicity and SH groups under identical conditions showed that blockage of SH groups by HpD binding is not leathal to the cells where as photoinduced cell death was observed on photodegradation of SH groups.