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. 2019 Aug 12:12:6361-6370.
doi: 10.2147/OTT.S210800. eCollection 2019.

The effect of MTHFD2 on the proliferation and migration of colorectal cancer cell lines

Yameng Wei #  1 , Pengfei Liu #  2 , Qian Li #  3 , Juan Du  1 , Yani Chen  1 , Yu Wang  1 , Haiyan Shi  1 , Yanfeng Wang  1 , Huahua Zhang  1 , Wanjuan Xue  1 , Yi Gao  1 , Dan Li  1 , Yun Feng  1 , Jing Yan  1 , Jiming Han  1 , Jing Zhang  1
Affiliations

The effect of MTHFD2 on the proliferation and migration of colorectal cancer cell lines

Yameng Wei et al. Onco Targets Ther. .

Abstract

Purpose: Methylenetetrahydrofolate dehydrogenase 2 (MTHFD2) is a tetramethylfolate dehydrogenase enzyme involved in folate metabolism. The aim of this study is to determine the effect of MTHFD2 on the proliferation and metastasis of colorectal cancer (CRC).

Patients and methods: MTHFD2 was silenced or overexpressed in CRC cells. qRT-PCR and Western blotting were used to analyze mRNA and protein expression, respectively. The MTT assay and colony forming assay were used to detect cell proliferation and colony formation ability. The cycle and apoptosis changes were detected by flow cytometry. Transwell experiments were used to analyze the migration ability of CRC cells.

Results: The expression of MTHFD2 in 31 kinds of cancers was analyzed by bioinformatics, and MTHFD2 was found highly expressed in various cancer cells including CRC cells. Silencing the expression of MTHFD2 resulted in inhibition of the proliferation of CRC cells, weakening of the migration ability, blocking of the cell cycle in G0/G1-S phase, and promotion of the apoptosis of CRC cells. On the contrary, overexpression of MTHFD2 in CRC cells resulted in enhancement of the proliferation and migration ability, promotion of cell cycle progression and inhibition of cell apoptosis.

Conclusion: MTHFD2 is positively related with colorectal cancer and the MTHFD2 gene is a tumor promoting gene in CRC cells.

Keywords: colorectal cancer; methylenetetrahydrofolate dehydrogenase 2; oncogene.

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Conflict of interest statement

The authors report no conflicts of interest in this work.

Figures

Figure 1
Figure 1
The expression of MTHFD2 in colorectal cancer (CRC) was up-regulated. (A) The expression of MTHFD2 in different types of human tumors within The Cancer Genome Atlas (TCGA). (B) The expression of MTHFD2 in CRC tissues (n=288) and normal tissues (n=35) in TCGA database. (C) The MTHFD2 mRNA and (D) protein expression levels were measured in CRC cell lines and a normal colon epithelial cell line (NCM-460). The intensity of each band was quantified, and β-actin was used as a housekeeping control (*P<0.05, **P<0.01, ***P<0.001, compared to the control group). Abbreviations: MTHFD2, Methylenetetrahydrofolate dehydrogenase 2; CRC, colorectal cancer.
Figure 2
Figure 2
Silencing MTHFD2 in CRC cells HCT116 and CACO-2 inhibited the proliferation and migration of cells. (A) HCT116 and caco-2 were transfected with si1- MTHFD2 and si2-MTHFD2, respectively. Silencing efficacy was detected with Western blot. (B) MTHFD2 mRNA and protein expression levels were measured in HCT116 and CACO-2 cells after transfection with si1-MTHFD2. (C) Colony-forming experiment showed the cell proliferative capacity of HCT116 and CACO-2 after transfection with si1-MTHFD2. (D) MTT assay analysis showed the cell growth of HCT116 and CACO-2 after transfection with si1-MTHFD2. (E) Transwell cell migration assay showed the cell migratory ability of HCT116 and CACO-2 after transfection with si1-MTHFD2. (**P<0.01, ***P<0.001, compared to the control group). Abbreviations: MTHFD2, Methylenetetrahydrofolate dehydrogenase 2; CRC, colorectal cancer.
Figure 3
Figure 3
Silencing MTHFD2 in CRC cells HCT116 and CACO-2 blocked the cell cycle in G0/G1-S phase, and promoted apoptosis. (A) The cell cycle was analyzed by flow cytometry in HCT116 and CACO-2 after transfection with si1-MTHFD2. (B) Western blot showed the expression of G0/G1-S-related genes. (C) Cell apoptosis was analyzed by flow cytometry in HCT116 and CACO-2 after transfection with si1-MTHFD2. (*P<0.05, **P<0.01, ***P<0.001, compared to the control group). Abbreviations: MTHFD2, Methylenetetrahydrofolate dehydrogenase 2; CRC, colorectal cancer.
Figure 4
Figure 4
Overexpression of MTHFD2 in CRC cell RKO and SW-480 promoted cell proliferation and migration. (A) MTHFD2 mRNA and protein expression levels were measured in RKO and SW-480 cells after transfection with the MTHFD2 vector. (B) Colony-forming experiment showed the cell proliferative capacity of RKO and SW-480 cells after overexpression of MTHFD2. (C) MTT assay analysis showed the cell growth of RKO and SW-480 cells after overexpression of MTHFD2. (D) Transwell cell migration assay showed the cell migration ability of RKO and SW-480 cells after overexpression of MTHFD2. (*P<0.05, **P<0.01, ***P<0.001, compared with the control group). Abbreviations: MTHFD2, Methylenetetrahydrofolate dehydrogenase 2; CRC, colorectal cancer.
Figure 5
Figure 5
Overexpression of MTHFD2 in CRC cell RKO and SW-480 inhibited cell apoptosis, and increased the M phase of cell cycle. (A) The cell cycle was measured by flow cytometry in RKO and SW-480 cells after transfection with the MTHFD2 vector. (B) Western blot showed the expression of G0/G1-S-related genes. (C) Cell apoptosis was measured by flow cytometry in RKO and SW-480 cells after the overexpression of MTHFD2. (*P<0.05, **P<0.01, ***P<0.001, compared with the control group). Abbreviations: MTHFD2, Methylenetetrahydrofolate dehydrogenase 2; CRC, colorectal cancer.
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