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. 2024 Feb 13;71:103087. doi: 10.1016/j.redox.2024.103087

Fig. 2.

Fig. 2

METTL17 is highly expressed in CRC and knockdown of METTL17 inhibits CRC cell proliferation, migration, invasion, and oncogenic signatures in vitro.

A. The mRNA levels of METTL17 were up-regulated in colon adenocarcinoma (COAD) compared to adjacent normal tissues based on TCGA analysis. Normal tissue (N), n = 41. Tumor tissues (T), n = 415.

B and C. The protein levels of METTL17 were measured in 12 pairs of human CRC tumor tissues (T) and the corresponding surrounding non-tumorous tissues (N) from 12 patients (abbreviated to P#1–12) (B). Relative protein level of METTL17 normalized to GAPDH was quantified by Image J (C). n = 12 per group. The METTL17 knockdown (#3) and Scramble (Sc) RKO cells were used as control.

D. METTL17 was highly expressed in human CRC cell lines compared to human normal colonic epithelial cell line (NCM460) detected by western blotting.

E. The knockdown efficiency of METTL17 in RKO, SW620, and DLD1 cells was confirmed by Western blot assays. Sc, Scramble. #3, shMETTL17#3. #5, shMETTL17#5.

F. Crystal violet staining assays showed that knockdown of METTL17 reduced cell proliferation of RKO, SW620, and DLD1. n = 5–6 per group. shM17#3, shMETTL17#3. shM17#5, shMETTL17#5.

G. Knockdown of METTL17 inhibited cell migration, invasion, and colony formation in RKO, SW620, and DLD1 cells measured through trans-well migration and invasion assays followed by crystal violet staining. The relative levels respectively were counted from Supplementary Fig. 3A, n = 4 per group.

H, I and J. Knockdown of METTL17 caused cell cycle arrest at G0/G1 phase in RKO cells (H) (n = 3 per group), and GSEA analysis revealed disruptions in cell cycle phase transition (I) and DNA replication (J) in METTL17-deficient RKO cells. GOBP, Gene Ontology Biological Process. FDR, false discovery rate.

K. GSEA analysis of RNA-Seq data identified that METTL17 knockdown in SW620 cells suppressed multiple oncogenic pathways.

L. Exogenous overexpression of METTL17 via lentivirus rescued the inhibited cell proliferation in METTL17-deficient SW620 cells. n = 5–6 per group. M17-WT, human wide type METTL17.

Data are shown as mean ± SD. ##p < 0.01, ###p < 0.001 compared to Scramble group, and **p < 0.01, ***p < 0.001 compared to Scramble group or the indicated two groups, based on two-sided Student's t-test or one-way ANOVA.