Effects of Glut1 gene silencing on proliferation, differentiation and apoptosis of colorectal cancer cells by targeting the TGF ‐β/PI3K‐AKT‐mTOR signaling pathway

This study aims to investigate the effects of glucose transport l (Glut1) gene on proliferation, differentiation and apoptosis of colorectal cancer (CRC) cells by regulating the TGF‐β/PI3K‐AKT‐mTOR signaling pathway. Immunohistochemistry was conducted to detect the positive Glut1 expression. Normal human CRC epithelial cells (CCD‐18Co) and CRC cell line HCT116 were grouped into the control, blank, negative control (NC), and shGlut1‐1 groups. RT‐qPCR and Western blotting were performed to detect the expressions of Glut1, TGF‐β1, PI3K, AKT, PTEN, mTOR, Bcl‐2 and Bax. Protein expression of phosphorylated‐PI3K (p‐PI3K), p‐S473‐AKT, p‐S389‐S6K1, p‐T70‐4EBP1, Cleaved caspase‐3 and Cleaved‐PARP were detected. MTT assay, flow cytometry and colony formation assay were performed in order to detect cell viability, cell cycle and apoptosis, respectively. The positive expression rate of Glut1 in CRC tissues was 75% ± 8%, while in the adjacent normal tissues it was 0%. In comparison to adjacent normal tissues, CRC tissues had increased Glut1, TGF‐β1, PI3K, AKT, mTOR and Bcl‐2 expressions, and p‐PI3K, p‐S473‐AKT, p‐S389‐S6K1 and p‐T70‐4EBP1 expressions; and decreased PTEN, Bax, Cleaved caspase‐3 and Cleaved‐PARP expressions. On comparison with the blank and NC groups, cells in the shGlut1‐1 group showed decreased Glut1, TGF‐β1, PI3K, AKT, mTOR and Bcl‐2 expressions, and p‐PI3K, p‐S473‐AKT, p‐S389‐S6K1 and p‐T70...
Source: Journal of Cellular Biochemistry - Category: Biochemistry Authors: Tags: Article Source Type: research