hydroxysafflor-yellow-a and Colorectal-Neoplasms

In recent years, the clinical treatment of colorectal cancer(CRC) has made great progress, but chemoresistance is still one of the main reasons for reducing the survival rate of patients with colorectal cancer. Therefore, ameliorating chemotherapy resis-tance is an urgent problem to be solved. The purpose of this study was to investigate the regulatory role and related molecular mechanisms of hydroxysafflor yellow A(HSYA) in colorectal cancer cell proliferation, migration, and 5-fluorouracil(5-FU) chemoresistance. In this study, HCT116 and HT-29 cells were used as research subjects. Firstly, methyl thiazolyl tetrazolium(MTT) assay and colony formation assay were used to detect and analyze the effect of HSYA on the proliferation of CRC cells. Secondly, the effect of HSYA on the cell cycle in CRC cells was analyzed by cell cycle assay. Furthermore, the effect of HSYA on the migration of CRC cells was analyzed by wound-healing assay and Transwell assay. Based on the above, the influences of HSYA on 5-FU chemoresistance of CRC cells and related molecular mechanisms were explored and analyzed. The results showed that HSYA significantly inhibited the proliferation and migration of CRC cells, and arrested the cell cycle in G_0/G_1 phase. In addition, HSYA significantly ameliorated the chemoresistance of CRC cells to 5-FU. The results of acridine orange staining and Western blot showed that the autophagy activity of CRC cells in the HSYA and 5-FU combined treatment group was significantly higher than that in the 5-FU single drug treatment group. As compared with the 5-FU single drug treatment group, the phosphorylation levels of protein kinase B(Akt) and mammalian target of rapamycin(mTOR) in the HSYA and 5-FU combined treatment group were significantly reduced, indicating that the Akt/mTOR signaling pathway in the combined treatment group was down-regulated in CRC cells. In conclusion, HSYA may upregulate autophagy activity through the Akt/mTOR signaling pathway, thereby inhibiting the proliferation and migration of CRC cells and ameliorating the chemoresistance to 5-FU.

Topics: Autophagy; Cell Line, Tumor; Cell Proliferation; Colorectal Neoplasms; Drug Resistance, Neoplasm; Fluorouracil; Humans; Proto-Oncogene Proteins c-akt; TOR Serine-Threonine Kinases

The natural compound Hydroxysafflor yellow A (HSYA) has been demonstrated to exert anti-cancer effect on multiple cancers. This study aimed to clarify the role of HSYA in inhibiting colorectal cancer (CRC) in vitro and the underlying mechanisms. Different concentrations of HSYA (0, 25, 50, and 100 μM) was exposed to HCT116 CRC cells, then cell proliferation, apoptosis, migration, and invasion were estimated by colony formation assay, TUNEL staining, wound-healing, and transwell assays, respectively. Western blotting assay was utilized to observe the expression of proteins involved in cell apoptosis, migration, and peroxisome proliferator-activated receptor γ (PPARγ)/PTEN/Akt signaling, including PCNA, Bax, Bcl-2, cleaved-caspase3, E-cadherin, N-cadherin, vimentin, PPARγ, and phosphorylated (p)-Akt. HCT116 cells that treated with 100 μM HSYA were also pre-treated with PPARγ antagonist, GW9662, or knockdown with PPARγ using short hairpin (sh)-RNA, to down-regulate PPARγ expression. Then, the above functional analysis was repeated. Results demonstrated that HSYA (25, 50 and 100 μM) significantly reduced HCT116 cell viability, but had no effect on the cell viability of human normal intestinal epithelial cell HIEC. HSYA also inhibited colony formation, migration, and invasion but promoted apoptosis of HCT116 cell in a concentration-dependent manner. Besides, the PPARγ/PTEN/Akt signaling was activated upon HSYA treatment. Finally, GW9662 and PPARγ knockdown blocked all the effects of HSYA on HCT116 cells. In conclusion, HSYA could exhibit anti-cancer effect on CRC via activating PPARγ/PTEN/Akt signaling, thereby inhibiting cells proliferation, migration, and invasion in vitro.

Topics: Apoptosis; Cell Line, Tumor; Cell Movement; Cell Proliferation; Chalcone; Colorectal Neoplasms; Down-Regulation; Humans; Neoplasm Invasiveness; PPAR gamma; Proto-Oncogene Proteins c-akt; PTEN Phosphohydrolase; Quinones; Signal Transduction