图1 结直肠癌组织中PIGT表达升高

Fig.1 Elevated expression of PIGT in colorectal cancer tissues. A: Pan cancer analysis based on the TNMplot database show that PIGT mRNA levels are significantly upregulated in multiple malignancies, including colorectal cancer (CRC), compared with corresponding normal tissues. B: Validation using the UALCAN database demonstrates that PIGT mRNA levels in primary tumor tissues are significantly higher than those in adjacent normal tissues in both colon adenocarcinoma (COAD) and rectal adenocarcinoma (READ). C: Analysis using the GEPIA database shows that PIGT mRNA expression is significantly increased in COAD and READ samples compared with normal controls. *P<0.05.

图2 PIGT高表达与中晚期结直肠癌患者不良预后相关

Fig.2 High PIGT expression is associated with poor prognosis in mid-to-late stage colorectal cancer patients.

图3 PIGT在结直肠癌细胞系中表达上调

Fig.3 PIGT expression is upregulated in colorectal cancer cell lines. A: qPCR analysis of PIGT mRNA levels in two colorectal cancer cell lines (HCT116 and HCT8) and the normal colorectal mucosal cell line NCM460. B: Western blotting analysis of PIGT protein expression in the same cell lines. All data represent the Mean±SD from 3 independent experiments. *P<0.05 vs NCM460.

图4 PIGT在HCT116和HCT8细胞中的表达被有效敲低

Fig.4 PIGT expression is efficiently knocked down in HCT116 and HCT8 cells (n=3). ***P<0.001.

图5 PIGT敲低抑制结直肠癌细胞增殖能力

Fig.5 Knockdown of PIGT inhibits proliferation of colorectal cancer cells. A: Knockdown of PIGT significantly reduces the number and size of colonies compared with the sh-NC control group. B: PIGT knockdown markedly decreases the number of colony spheres formed in soft agar. ***P<0.001 (n=3).

图6 PIGT敲低抑制结直肠癌细胞的迁移与侵袭能力

Fig.6 Knockdown of PIGT attenuates migration and invasion of CRC cells. A: Wound healing assay in HCT116 and HCT8 cells transfected with sh NC or sh PIGT. Knockdown of PIGT significantly delays wound closure compared with the control group. B: Transwell migration assay. PIGT knockdown markedly reduces the number of cells migrating through the polycarbonate membrane. C: Matrigel invasion assay. PIGT knockdown similarly impairs the ability of cells to penetrate the Matrigel barrier. ***P<0.001 (n=3).

图7 PIGT敲低抑制结直肠癌细胞的血管形成能力

Fig.7 Knockdown of PIGT impairs tube formation in CRC cells (n=3). ***P<0.001.

图8 PIGT敲低可下调效应分子表达,从而逆转CRC细胞的EMT

Fig.8 PIGT knockdown reverses EMT by downregulating its effector molecules in CRC cells. A: Western blotting of N cadherin, E cadherin, Vimentin, MMP9, and VEGF-C protein levels in HCT116 cells after PIGT knockdown. GAPDH was used as a loading control. B: Western blotting and quantification of the same proteins in HCT8 cells. ***P<0.001 (n= 3).

图9 功能富集分析揭示了与PIGT相关的关键通路与生物学过程

Fig.9 Functional enrichment analysis reveals key pathways and biological processes associated with PIGT. A: Number of differentially expressed genes upon PIGT knockdown. B: GO enrichment analysis of the differentially expressed genes. C: KEGG pathway enrichment analysis of the differentially expressed genes.

图10 PIGT调控NF-κB信号通路的蛋白质水平验证

Fig.10 Protein level validation of PIGT-mediated regulation of NF‑κB signaling. A: Western blotting of P65 and phosphorylated P65 (P-P65) in HCT116 cells after PIGT knockdown. B: Western blotting analysis of P65 and P-P65 in HCT8 cells after PIGT knockdown. ***P<0.001 (n=3).

图11 脂多糖逆转 PIGT 敲低对结直肠癌细胞 NF-κB 通路及 EMT 的抑制作用

Fig.11 LPS reverses PIGT knockdown-induced inhibition of NF-κB pathway and EMT in colorectal cancer cells. A: Western blotting of P P65, E cadherin, N cadherin, vimentin, MMP9, and VEGF-C in HCT116 cells in sh NC, sh PIGT, and sh PIGT+LPS (1 μg/mL LPS, 24 h) groups. B: Western blotting analysis of the same proteins in HCT8 cells. *P<0.05, **P<0.01, ***P<0.001 (n=3).

图 12 敲低 PIGT 抑制裸鼠皮下结直肠癌肿瘤生长

Fig.12 Knockdown of PIGT inhibits growth of colorectal cancer xenografts in nude mice. A: Representative images of xenograft tumors derived from HCT116 cells transfected with sh NC or sh PIGT. B: Western blotting of PIGT protein expression in tumor tissues, confirming efficient PIGT knockdown in vivo. **P<0.01, ***P<0.001.