Activation of P27kip1-cyclin D1/E-CDK2 pathway by polysaccharide from Phellinus linteus leads to S-phase arrest in HT-29 cells

• Published in: Chemico-biological interactions Vol. 206; no. 2; pp. 222 - 229
• Main Authors: Zhong, Shi, Ji, Dong-Feng, Li, You-Gui, Lin, Tian-Bao, Lv, Zhi-Qiang, Chen, Hua-Ping
• Format: Journal Article
• Language: English
• Published: Ireland Elsevier Ireland Ltd 25.11.2013
• Subjects: alanine transaminase; Animals; apoptosis; aspartate transaminase; autophagy; blood serum; body weight; Caco-2 Cells; cell cycle; Cell Line; cell proliferation; Cell Proliferation - drug effects; colorectal neoplasms; Cyclin D1 - metabolism; Cyclin E - metabolism; cyclin-dependent kinase; Cyclin-Dependent Kinase 2 - metabolism; Cyclin-Dependent Kinase Inhibitor p27 - metabolism; Cyclins/CDK complexes; Fungi - chemistry; gene expression regulation; HT29 Cells; Humans; kidneys; liver; mechanism of action; Mice; P27kip1; Phellinus linteus; Polysaccharide; Polysaccharides - chemistry; Polysaccharides - isolation & purification; Polysaccharides - pharmacology; proteins; reverse transcriptase polymerase chain reaction; S Phase Cell Cycle Checkpoints - drug effects; S-phase arrest; Signal Transduction - drug effects; toxicity; Western blotting; P27kip1; Cyclins/CDK complexes; Polysaccharide; S-phase arrest
• ISSN: 0009-2797; 1872-7786
• DOI: 10.1016/j.cbi.2013.09.008

P1 induced S phase arrest in HT-29 cells. •Proteoglycan (P1) from Phellinus linteus inhibits HT-29 cell proliferation in vitro and in vivo.•P1-induced S phase arrest but no apoptosis and autophagy in HT-29 cells.•P1 activated P27kip1-cyclin D1/E-CDK2 pathway in HT-29 cells. Our previous study showed that polysaccharide (P1) from Phellinus linteus exhibits a significant inhibitive activity on human colorectal carcinoma cells (HT-29). However its novel molecular mechanism remains unknown. To obtain insights into P1’s mechanism of action, we examined its effects on cell proliferation in vitro and in vivo, cell cycle distribution, apoptosis, autophagy, and expression of several cell cycle interrelated proteins in HT-29 cells. Interestingly, we found that volume and weight of the solid tumor significantly decreased in P1 (200mg/kg)-treated mice compared with the control. However, slightly increased the body weight of the P1 treated tumor-bearing mice, with no significant increased ALT, AST levels in serum and LPO concentration in liver and kidney indicated that P1 has no toxicity to mammals at a dose of 200mg/kg. Furthermore, P1 caused a significantly dose-dependent increase in the S-phase cell cycle, but no apoptosis and autophagy in HT-29 cells. RT-PCR and Western blot results showed significantly down-regulated expressions of cyclin D1, cyclin E, and CDK2, as well as increased expressions of P27kip1 in P1 (100μg/mL)-treated HT-29 cells. These results suggested that the activation of P27kip1-cyclin D1/E-CDK2 pathway is involved in P1-induced S-phase cell cycle arrest in HT-29 cells.