Matrine promotes colorectal cancer apoptosis by downregulating shank-associated RH domain interactor expression

Figure 1 Flow chart. CRC: Colorectal cancer; IHC: Immunohistochemistry.

Figure 2 Proteomic profiles and bioinformatic analysis of colorectal cancer patients. A: Hierarchical cluster analysis shows the global differentially expressed proteins. Upregulated (red) and downregulated (green) differentially expressed proteins in the CRC group compared to the normal group; B: Principal component analysis shows the protein expression in the cancer tissues (green) and normal tissues (orange) of colorectal cancer (CRC) patients; C: Volcano plot shows the fold changes of identified proteins. Significantly upregulated or downregulated proteins are represented in red and blue, respectively, while proteins without significant changes are shown in gray; D: Gene Ontology analysis shows the enrichment results for differentially expressed proteins; E: Kyoto encyclopedia of genes and genomes analysis shows that differentially expressed proteins are significantly enriched in multiple pathways, ^cP < 0.001. CRC: Colorectal cancer; PCA: Principal component analysis; HCA: Hierarchical cluster analysis; GO: Gene ontology; KEGG: Kyoto encyclopedia of genes and genomes; PI3K-Akt: Phosphatidylinositol 3-kinase serine/threonine kinase; ECM: Extracellular matrix.

Figure 3 The expression of shank-associated RH domain interactor, metalloproteinase inhibitor 1, nucleolar complex protein 2 homolog, and DNA ligase 4 proteins and its association with tumor-node-metastasis and overall survival in colorectal cancer. A: The expression of 4 proteins among tumor-node-metastasis (TNM) stages in 52 colorectal cancer (CRC) patients; B: The expression of 4 proteins in TCGA; C: The expression of 4 proteins among TNM stages in TCGA; D: The association between expression of 4 proteins and overall survival in TCGA; E: The immunohistochemistry of shank-associated RH domain interactor in 93 CRC tissues and 35 normal tissues. ^aP < 0.05; ^bP < 0.01; CRC: Colorectal cancer; HC: Health control; SHARPIN: Shank-associated RH domain interactor; TIMP1: Metalloproteinase inhibitor 1; NOC2 L: Nucleolar complex protein 2 homolog; LIG4: DNA ligase 4.

Figure 4 Matrine induces apoptosis and regulates apoptotic proteins by downregulating shank-associated RH domain interactor. A: The apoptosis rate of colorectal cancer (CRC) cells in 4 groups [control group (con group), SHARPIN knockdown group (siSHARPIN group), matrine, siSHARPIN + matrine]; B: The expression of caspase 3, caspase 8, B-cell lymphoma-2 (Bcl-2) associated X protein, Bcl-2 associated agonist of cell death, Bcl-2 proteins in CRC cells in 4 groups; C: The suppressing effect of matrine on shank-associated RH domain interactor (SHARPIN) in CRC cells. ^aP < 0.05; ^bP < 0.01; ^cP < 0.05 vs con,^dP < 0.01 vs con; ^eP < 0.05 vs siSHARPIN,^fP < 0.01 vs siSHARPIN; ^gP < 0.05 vs matrine,^hP < 0.01 vs matrine; Con: Control group; siSHARPIN: SHARPIN knockdown group; matrine: Matrine group; siSHARPIN + matrine: Matrine and SHARPIN knockdown group; CRC: Colorectal cancer; SHARPIN: Shank-associated RH domain interactor; Bcl-2: B-cell lymphoma-2; BAD: Bcl-2 associated agonist of cell death; BAX: Bcl-2 associated X protein.

Figure 5 Matrine impedes proliferation, invasion and migration in colorectal cancer by downregulating shank-associated RH domain interactor. A: The viability of colorectal cancer (CRC) cells using the cell counting kit-8 assay in 4 groups [control (con), si-Shank-associated RH domain interactor (SHARPIN), matrine, siSHARPIN + matrine]; B: The invasion cells of CRC in 4 groups; C: The wound healing of CRC cells in 4 groups. ^cP < 0.05 vs con,^dP < 0.01 vs con; ^eP < 0.05 vs siSHARPIN,^fP < 0.01 vs siSHARPIN; ^gP < 0.05 vs matrine,^hP < 0.01 vs matrine; Con: Control group; SHARPIN: Shank-associated RH domain interactor; siSHARPIN: SHARPIN knockdown group; matrine: Matrine group; siSHARPIN + matrine: Matrine and SHARPIN knockdown group; CRC: Colorectal cancer.