Fig. 1

Expression of TMCO1 mRNA in cervical cancer and its relationship with survival A: Differential expression of TMCO1 in cervical cancer and adjacent normal tissues. B: Survival curves of different TMCO1 mRNA expression levels in cervical cancer."

Fig. 2

Overexpression of TMCO1 promotes the proliferation of cervical cancer HeLa cells A: A TMCO1 lentiviral expression vector was infected with the cervical cancer cell line HeLa to obtain a cell line stably expressing TMCO1. The cell line infected with the EV was used as a control, and the expression of TMCO1 was confirmed by IB. B: The stably transfected TMCO1 cells and control cells (EV) were subjected to CCK-8 cell proliferation assay. C, D: The stably transfected TMCO1 cells and control cells (EV) were seeded into 6-well plates at 1 000 cells/well, and the number of clones formed was counted after 10 days of culture. E, F: The stably transfected TMCO1 cells and control cells (EV) were labeled with EdU to detect the number of cells undergoing active DNA synthesis. *: P<0.05, **: P<0.01. EV: Empty vector."

Fig. 3

Knocking down TMCO1 inhibited the proliferation of cervical cancer HeLa cells A: Two independent shRNAs targeting the TMCO1 gene sequence were designed and cloned into the pLKO.1 lentiviral vector, which was used to infect the cervical cancer cell line HeLa. After puromycin selection, cells with stable knockdown of TMCO1 were obtained, and cells infected with the empty vector (pLKO.1) virus were used as controls. The cells were collected and the knockdown of TMCO1, as well as the phosphorylation of cell cycle inhibitory protein p27 and histone H3 were detected by IB. B, C: The above TMCO1 stable knockdown cells and control cells were seeded into 6-well plates at 1 000 cells/well, and the number of clones formed was counted after 10 days of culture. D, E: The above TMCO1 stable knockdown cells and control cells were EdU labeled to detect the number of cells undergoing active DNA synthesis. *: P<0.05, **: P<0.01, ***: P<0.001, compared with empty vector (pLKO.1)."

Fig. 4

TMCO1 promoted the migration of cervical cancer HeLa cells A, B: The stably transfected TMCO1 cells and control cells (EV) obtained in Figure 2 were inoculated into chambers according to the experimental steps for transwell cell migration assay. After 24 hours, the cells were fixed with formaldehyde and stained with crystal violet, and the number of migrated cells was observed and photographed under a microscope and the number of migrated cells was counted. C, D: The TMCO1 stable knockdown cells and control cells (pLKO.1) in Figure 3 were inoculated into the chamber according to the experimental steps for the transwell cell migration experiment. After 24 hours, the cells were fixed with formaldehyde and stained with crystal violet. The number of migrated cells was observed and photographed under a microscope, and the number of migrated cells was counted. ***: P <0.001, compared with empty vector (EV or pLKO.1)."

Fig. 5

TMCO1 regulated cell adhesion and signaling transduction A: The Flag-TMCO1 cells and control cells (EV) in Figure 2 were collected, cell lysates were prepared and trypsinized, and then analyzed by liquid chromatography-tandem mass spectrometry. Data were collected for principal component analysis, and it can be seen that the two Flag-TMCO1 stable expression cell line samples and the three control cell (EV) samples were clustered together, indicating that the stable expression of TMCO1 caused significant changes in the proteomic level. B: The differentially expressed proteins obtained by quantitative analysis of the protein expression profile data obtained in (A) were analyzed and a volcano plot was drawn, and representative proteins with significant differences were annotated. C, D: GSEA-KEGG pathway enrichment analysis was performed on the differentially expressed proteins in the Flag-TMCO1 stable expression cells and control cells (EV) protein expression groups, showing that pathways related to extracellular matrix-adhesion and PI3K-AKT signaling were significantly upregulated in Flag-TMCO1 stable expression cells, while ribosome-related pathways were downregulated."