Abstract
BACKGROUND The aims of this study were to investigate the expression of methylenetetrahydrofolate dehydrogenase 1 (MTHFD1) in human tissue containing clear cell renal cell carcinoma (CCRCC) compared with normal renal tissue, and the effects of upregulating the expression of MTHFD1 in the human CCRCC cell line, Caki-1. MATERIAL AND METHODS Tumor and adjacent normal renal tissue were obtained from 44 patients who underwent radical nephrectomy for CCRCC. Caki-1 human CCRCC cells were divided into the control group, the empty vector (EV) group, and the plasmid-treated group that overexpressed MTHFD1. MTHFD1 mRNA and protein levels were measured by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot, respectively. The cell counting kit-8 (CCK-8) assay measured cell viability. Flow cytometry evaluated apoptosis and the cell cycle. Western blot measured the protein levels of MTHFD1, Bax, Bcl-2, Akt, p53, and cyclin D1, and qRT-PCR determined the gene expression profiles. RESULTS MTHFD1 mRNA and protein levels in CCRCC tumor tissues were significantly lower compared with adjacent normal renal tissue. MTHFD1 over-expression in Caki-1 cells inhibited cell proliferation, arrested cells in the G1 phase, increased cell apoptosis, and upregulated gene and protein expression of Bax/Bcl-2 and p53 and inhibited p-Akt, and cyclin D1. CONCLUSIONS MTHFD1 was underexpressed in CCRCC tissue when compared with normal renal tissue. MTHFD1 transfection of human CCRCC Caki-1 cells in vitro inhibited cell proliferation and promoted apoptosis, associated with reduced expression of cyclin D1, reduced Akt phosphorylation, and increased expression of Bax/Bcl-2 and p53.
MeSH terms
-
Apoptosis / physiology
-
Carcinoma, Renal Cell / enzymology*
-
Carcinoma, Renal Cell / genetics
-
Carcinoma, Renal Cell / metabolism
-
Carcinoma, Renal Cell / pathology
-
Cell Cycle / physiology
-
Cell Line, Tumor
-
Cell Movement / physiology
-
Cell Proliferation / physiology
-
Cyclin D1 / biosynthesis
-
Cyclin D1 / genetics
-
Cyclin D1 / metabolism
-
Humans
-
Kidney Neoplasms / enzymology*
-
Kidney Neoplasms / genetics
-
Kidney Neoplasms / metabolism
-
Kidney Neoplasms / pathology
-
Methylenetetrahydrofolate Dehydrogenase (NADP) / biosynthesis*
-
Methylenetetrahydrofolate Dehydrogenase (NADP) / genetics
-
Methylenetetrahydrofolate Dehydrogenase (NADP) / metabolism
-
Minor Histocompatibility Antigens / biosynthesis*
-
Minor Histocompatibility Antigens / genetics
-
Minor Histocompatibility Antigens / metabolism
-
Phosphorylation
-
Proto-Oncogene Proteins c-akt / biosynthesis
-
Proto-Oncogene Proteins c-akt / genetics
-
Proto-Oncogene Proteins c-akt / metabolism
-
Proto-Oncogene Proteins c-bcl-2 / biosynthesis
-
Proto-Oncogene Proteins c-bcl-2 / genetics
-
Proto-Oncogene Proteins c-bcl-2 / metabolism
-
RNA, Messenger / genetics
-
RNA, Messenger / metabolism
-
Transcriptome
-
Tumor Suppressor Protein p53 / biosynthesis
-
Tumor Suppressor Protein p53 / genetics
-
Tumor Suppressor Protein p53 / metabolism
-
bcl-2-Associated X Protein / biosynthesis
-
bcl-2-Associated X Protein / genetics
-
bcl-2-Associated X Protein / metabolism
Substances
-
BAX protein, human
-
BCL2 protein, human
-
CCND1 protein, human
-
Minor Histocompatibility Antigens
-
Proto-Oncogene Proteins c-bcl-2
-
RNA, Messenger
-
TP53 protein, human
-
Tumor Suppressor Protein p53
-
bcl-2-Associated X Protein
-
Cyclin D1
-
MTHFD1 protein, human
-
Methylenetetrahydrofolate Dehydrogenase (NADP)
-
Proto-Oncogene Proteins c-akt