Trypsin

Endostar was provided by Simcere Pharmaceutical Group (Nanjing, China). RPMI-1640 medium and trypsin were provided by Nanjing KeyGen Biotech Co., Ltd. (Nanjing, China). Fetal bovine serum (FBS) was purchased from Gibco (Thermo Fisher Scientific, Inc., Waltham, MA, USA). Antibodies against CD31 (cat. no. ab28364), hypoxia-inducible factor-1α (HIF-1α; cat. no. ab2185), MMP-2 (cat. no. ab37150), MMP-9 (cat. no. ab38898), VEGF (cat. no. ab46154), IFN-γ (cat. no. ab198801), IL-17 (cat. no. ab79056) and CD86 (cat. no. ab119857) were provided by Abcam (Cambridge, MA, USA). Mouse ELISA kits for tumor necrosis factor-α (TNF-α; MK1169), IFN-γ (MK1205), IL-17 (MK1445), α-fetoprotein (AFP; MK2689) and cyclic adenosine 5′-phosphate (cAMP; MK2893) were ordered from Wuhan Boster Biological Technology, Ltd. (Wuhan, China). A monoclonal β-actin antibody (sc-47778) was bought from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). The secondary antibody (horseradish peroxidase conjugated-anti-rabbit IgG) was obtained from Invitrogen (A-11034; Thermo Fisher Scientific, Inc.).

Hydrogen peroxide (China National Medicines Co. Ltd., Shanghai, China), the cytotoxicity and caspase-3 activity detection kit (Biyuntian Institute of Biotechnology, Nantong, China), the electrochemiluminescence (ECL) immunoblotting substrate kit (Merck Millipore, Burlington, MA, USA), antibody to DK2 (Abcam, Cambridge, MA, USA), microplate reader (Bio-Rad, Hercules, CA, USA), 1640 medium and fetal bovine serum (FBS) (Gibco, Thermofisher Scientific, Waltham, MA, USA), and trypsin, the Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) cell apoptosis detection kit (Nanjing KeyGen Biotech Co. Ltd., Nanjing, China) were obtained from the respective companies. The other reagents were of analytical grade purity. A 680 full-automatic microplate reader (Bio-Rad, Hercules, CA, USA), an AE31EF-INV inverted microscope (Motic Microscopy, Tokyo, Japan), a CO₂ cell incubator (Sanyo Co., Tokyo, Japan), and a C6 flow cytometer (Beckman Coulter, Brea, CA, USA) were used for the assays.

Human cervical cancer HeLa cells were maintained in high-glucose Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum, and were incubated in a 37°C incubator with a 5% CO₂ atmosphere, under saturated humidity (2406-2 CO₂ incubator; Heraeus, Hanau, Germany). A high pressure sterilization autoclave (ES 315; Tomy Company, Ltd., Tokyo, Japan was used to prepare the phosphate-buffered saline (PBS) buffer solution for cell passage cultivation. The cells were passaged with 0.25% trypsin (Nanjing KeyGen Biotech Co., Ltd.). The cells used in the present study were in the logarithmic phase of growth at 24 h subsequent to incubation. β-HIVS (2.3 mg) was dissolved in dimethyl sulfoxide (592 µl) and the product (10 mM) was stored at −20°C prior to use. A thermostatic bath (W201B; Senco Technology Co., Ltd., Shanghai, China) was used to rewarm the frozen cell solutions.

N, N-Dimethylformamide (DMF) was dried overnight with calcium hydride (98%, Aladdin), then distilled to use. Dasatinib (DAS) (99%) and O-(1H-benzotriazol-1-yl)- N,N,N',N'-tetramethyluronium tetrafluorob-orate (TBTU) (98%) and sodium diethyldithiocarbamate (98%) were obtained from Aladdin (Shanghai, China). Cisplatin (CP) was obtained from Shandong platinum source chemical Co. Ltd (Jinan, China). Anhydrous ethanol, hydrogen peroxide (30%), anhydrous ether, DMF, triethylamine (TEA), succinic anhydridedichloromethane (DCM), anhydrous methanol and pyridine were all purchased from Sinopharm Chemical Reagent Co., Ltd (Shanghai, China). Cell culture medium, trypsin and fetal bovine serum were bought from KeyGEN BioTECH (Nanjing, China). Poly(2-hydroxyethyl methacrylate) (PHEMA) and 3-(4, 5-dimethylthiazol-2-yl)−2, 5-diphenyl tetrazolium bromide (MTT) were bought from Sigma-Aldrich. Male ICR mice (SPF, 18–22 g) were obtained from Animal Center of Anhui Medical University (Hefei, China).