Bel 7402 cells

Bel-7402 cells were purchased from the American Type Culture Collection (ATCC; USA) and cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (Invitrogen, USA) containing 10% fetal bovine serum (Sigma, USA), 100 U/mL penicillin, and 50 μg/mL streptomycin in 20 cm² tissue-culture flasks (Falcon, USA). Cells were maintained at 37°C in humidified incubators with 5% CO₂ and passaged every 3 days.
New Zealand rabbits (weight 1.8–2.5 kg), imprinting control region (ICR) mice (weight 18–22 g, males and females of equal number) and Sprague Dawley rats (weight 180–200 g) were purchased from the Shanghai Laboratory Animal Center, Chinese Academy of Sciences, People’s Republic of China (PRC). Six-week-old nude male BALB/c mice (weight 18–20 g; Shanghai Laboratory Animal Center) were housed in laminar flow cabinets under specific pathogen-free conditions and provided with water and food ad libitum. All animals were cared for and handled according to the recommendations of the National Institutes of Health’s Guide for the Care and Use of Laboratory Animals. Animal protocols were approved by the Shanghai Medical Experimental Animal Care Committee.

HepG2 cells and Bel-7402 cells (American Type Culture Collection, Manassas, VA, USA) were cultured in RPMI-1640 medium containing 10% fetal bovine serum in 5% CO₂ at 37°C as a monolayer culture. As previously described,^{10 (link)–12 (link, link)} a liquid overlay technique was used to obtain tumour multicellular spheroids. Briefly, a cell suspension was seeded at 2 x 10⁵ cells in each culture flask coated with 2% agarose (Sigma-Aldrich, St Louis, MO, USA) before cell plating. Tumour multicellular spheroids were obtained after incubation for 4 days and the formation process of multicellular spheroids was observed by means of optical microscopy (AE2000LED inverted microscope; Motic, Xiamen, China).

Tetraethyl orthosilicate (TEOS, 99 wt.%), cetyltrimethylammonium bromide (CTAB), N,N’-dicyclohexylcarbodimide (DCC, >98%), 2-N-morpholino-ethanesulfonic acid (MES), 4-dimethylaminopyridine (DMAP) and N-hydroxysuccinimide (NHS, >98%) were purchased from Aladdin. 2-cyanoethyltriethoxysilane (CETES), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide-HCl (EDC, >98%), epsilon-aminocaproic acid (ACA), 2-hydroxyethyl trimethylammonium chloride (ChCl), Fluorescein isothiocyanate (Flu) and acetylcholine esterase (AChE) were purchased from Sigma-Aldrich. p-Sulfonatocalix[6]arene (p-SC₆A) was obtained from TCI Development Co., Ltd. (Shanghai, China). 1,2,3,4-Tetrahydro-5-aminoacridine hydrochloride (Tacrine, Tac) was purchased from Heowns Biochem. All these reagents were used as received without further purification. Deionized water (18.2 MΩ cm) used for all experiments was obtained from a Milli-Q system (Millipore, Bedford, MA).
C57BL6/J mice were obtained from the Experimental Animal Center of the Chinese Academy of Medical Sciences. Bel7402 cells were obtained from American Type Culture Collection. RPMI 1640 and fetal bovine serum (FBS) were purchased from Gibco BRL. (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was bought from Sigma-Aldrich.