Table 1.
The primers related to FAM50A.
Fig 1.
High expression of FAM50A in CRC.
(A) Panoramic analysis of FAM50A gene expression in tumors and normal tissues (unpaired) from the TCGA database. (B) Panoramic analysis of FAM50A gene expression in tumors and corresponding normal tissues (paired). (C) Expression of FAM50A in tumor tissues and normal tissues (unpaired). (D) Expression of FAM50A in paired tumor and normal tissues. (E) IHC showed high expression of FAM50A in CRC tumor cells (upper panel: 40X; lower panel: 200X). (F) Normal cells showed only weak staining for FAM50A (upper panel: 40X; lower panel: 200X). * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, ns: not significant.
Table 2.
Patient baseline data involving correlation between the clinical characteristics and pathological staging in CRC patients.
Fig 2.
Correlation of FAM50A expression with clinical and pathological features of CRC.
(A) Expression of FAM50A in normal and cancer tissues. (B) ROC curve analysis identified the different expression between the two groups. (C) Expression of FAM50A in male and female patients with CRC. (D) Expression of FAM50A in different age groups. (E) Expression of FAM50A in relation to tumor size. (F) Expression of FAM50A in relation to tumor location. (G) FAM50A expression in relation to cancer site. (H) FAM50A expression in relation to histological differentiation (M, moderate differentiation; P, poor differentiation). (I) FAM50A expression in relation to the depth of tumor invasion. (J) FAM50A expression according to lymph node metastasis. (K) FAM50A expression according to the presence of distant metastasis. (L) FAM50A expression according to TNM stage. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, ns: not significant.
Fig 3.
Cox regression analysis in CRC.
(A) Overall survival rate in CRC patients (n = 145). (B) OS in female and male groups. (C) OS in different age groups. (D) OS related to tumor size. (E) OS related to tumor location. (F) OS of ddifferent cancer sites. (G) OS interrelated with differentiation. (H-K) OS in T, N, M and TNM stages. (L) OS in FAM50A high and low groups. * P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001 and ns: no significance.
Table 3.
Univariate and multivariate Cox proportional hazards regression analysis in CRC patients.
Fig 4.
Forest plot of prognostic risk factors for CRC.
(A) Univariate Cox regression analysis forest plot. (B) Multivariate Cox regression analysis forest plot.
Fig 5.
Nomogram to display the prognosis value of FAM50A in CRC.
(A-E) represent the results of prognostic proportional risk analysis in T, N, M and TNM stages and FM50A expression. (F) Nomogram constructed with different variables. (G) Calibration curve of nomogram.
Fig 6.
Knockdown of FAM50A inhibited CRC cells proliferation ability in vitro.
(A) Detection of FAM50A expression in CRC tumor cell lines and normal cell by using RT-qPCR and WB. (B) Verification of knockdown efficiency in SW480. (C) Verification of knockdown efficiency in HCT-8. (D) Detection of cell proliferation ability by CCK-8 assays in SW480. (E) CCK-8 assays in HCT-8. (F) EdU staining to identify the changes in cell proliferation after FAM50A knockdown in SW480 and HCT-8. (G) Plate colony formation assay to identify changes in cell proliferation in FAM50A knockdown cells. sh means knockdown, * P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001 and ns: no significance.
Fig 7.
Overexpression of FAM50A promoted cell proliferation ability in RKO.
(A) Evaluation of the high expression efficiency for FAM50A. (B) CCK-8 assays in RKO. (C) EdU stained cells in overexpression cell compared with control RKO cell. (D) The colony forming ability assay of RKO.
Fig 8.
Knocking down FAM50A leads to S phase arrest in CRC cells.
(A-B) Cell cycle assays were performed on SW480 and HCT-8 cells after FAM50A was knocked down (up: SW480, down: HCT-8). (C-D) Western blot assays were conducted to detect the expression of cell cycle and cell cycle-related proteins in CRC cells.