Prmi 1640

The human OSCC cell lines used in this study were HN6, OSC4, SCC15, SCC25 and CAL27. The human gastric adenocarcinoma cell line (MGC803) expressing CREPT stably was applied as positive control [6 (link)], and the normal human oral keratinocyte cell line (HOK) as normal control[10 (link), 20 (link), 21 ]. SCC25, CAL27 and HOK were obtained from the American type culture collection (ATCC). HN6 and SCC15 were kindly provided by Beijing Institute of Dental Research, Stomatological Hospital and School of Stomatology.MGC803 and OSC4 weregifted by Department of Gastroenterology & Hepatology, Chinese PLA General Hospital. The HN6, SCC15 and CAL27 cells were incubated in DMEM medium (Hyclone, Logan, UT) containing 10% fetal bovine serum (FBS; Gibco, USA). SCC25 cells were grown in DMEM/Ham’s F-12 medium supplemented with 20% FBS (HyClone, Logan, USA), hydrocortisone (40 ng/mL) and sodium pyruvate (1 mM). The OSC4 and MGC803 cells were incubated in PRMI-1640 (Gibco, USA) containing 10% FBS (Gibco, USA). The HOK cells were incubated in oral keratinocyte medium (OKM; ScienCell Research Laboratories, Carlsbad, CA.USA) containing growth supplements as previously described [21 , 22 (link)].

The human breast cancer cell lines are BT-549, MCF-7, MDA-MB-231, MDA-MB-468, SK-BR-3, T47D, YCC-B1, and ZR-75-1, normal mammary epithelial cell lines HMEC and MCF-10A were acquired from American Type Culture Collection. Breast cancer cell lines were cultured in PRMI-1640 (Gibco BRL, Karlsruhe, Germany) supplemented with 10% fetal bovine serum (FBS) (Invitrogen, Carlsbad, CA), 100 U/mL penicillin, and 100 μg/mL streptomycin (Gibco BRL, Karlsruhe, Germany) at 37°C with 5% CO₂. The tamoxifen-resistant MCF-7 (MCF-7R) breast cancer cell was a gift from Dr. Lin29 (link) (Shantou Affiliated Hospital of Sun Yat-Sen University) and was cultured in RPMI-1640 without phenol red and L-glutamine (Biological industries, Haemek, Israel) containing 10% charcoal-stripped FBS (Biological industries, Haemek, Israel) and 1 μM 4-hydroxytamoxifen (Sigma-Aldrich, MO, USA). Human breast cancer specimens and paired normal tissues were collected at the First Affiliated Hospital of Chongqing Medical University. Each patient provided informed consent. The acquisition of human specimens was authorized by the Institutional Ethics Committee of the First Affiliated Hospital of Chongqing Medical University.

Bone marrow from leukemia patients and healthy donors was obtained after informed consent in accordance with the guidelines of the first hospital affiliated to Xiamen university, and the study protocols were approved by the Ethics Committee. Human leukemia cell lines K562, Kasumi-1, and THP-1, and human embryonic kidney cell line 293T were used in this study and maintained, respectively, in PRMI 1640 and DMEM medium (Gibco, Carlsbad, CA, USA) supplemented with 10% heat-inactivated fetal bovine serum, l-glutamine, and antibiotics at 37 °C in 5% CO₂.

Lithium chloride (LiCl, a conventional GSK-3β inhibitor) was purchased from Sigma. Human colon cancer cell line SW480 was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). Cells were maintained in PRMI 1640 (Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (Gibco), 100 U/mL penicillin, and 100 μg/mL streptomycin (Invitrogen) and kept in a humidified atmosphere at 37°C with 5% CO₂ in air.

The human OSCC cell lines used were Tca8113, OSC-4, SCC1, SCC2, SCC4, SCC9, CAL-27, UM1, and UM2. The human hepatocellular carcinoma cell lines used were SMMC7721, HCCLM3, HUH7, and HEPG2. The human breast cancer cell lines used were MCF7, BT474, SKBR3, HCC1187, and HCC1143. The normal human liver cell line LO2, the normal human oral keratinocyte cell line HOK, and the normal human breast cell MCF-10A were purchased from American type culture collection (Manassas, VA, USA). The Tca8113, CAL-27, MCF 10A, SMMC7721, HCCLM3, HEPG2, and BT474 cells were incubated in DMEM medium (Hyclone, Logan, UT) containing 10 % FBS (Invitrogen, Carlsbad, CA) at 37 °C with 5 % CO2. The OSC-4, SCC2, SCC9, LO2, HCC1143, and SKBR3 cells were incubated in PRMI-1640 (Gibco, American) containing 10 % FBS. The SCC-4, MCF7, and HUH7 cells were incubated in DMEM: F12 containing 10 % FBS. The HOK cells were incubated in OKM containing growth factor. Approximately 5 % of Tca8113 cells were plated into each well of 12-well plates at least 24 h before transfection to achieve 30–50 % confluency.

Human embryonic kidney 293T (HEK-293T) cells and human leukemic monocyte (THP-1) cells were obtained from the Institute of Medical Biology (Kunming, China) and propagated in Dulbecco′s modified Eagle′s medium (DMEM, Thermo Fisher Scientific, MA, USA), PRMI 1640 (Gibco, MA, USA), respectively, supplemented with 10% (v/v) fetal bovine serum (FBS, Gibco) and penicillin–streptomycin (100 U/mL and 100 µg/mL). African green monkey kidney (Vero) cells obtained from the Institute of Medical Biology and human foreskin fibroblast (HFF) cells were purchased from Cell Resources Center, Shanghai Academy of Sciences, Chinese Academy of Sciences (Shanghai, China) and cultured in minimum Eagle′s medium (MEM, Thermo Fisher Scientific, MA, USA), supplied with 10% FBS and penicillin–streptomycin (100 U/mL and 100 µg/mL). The cells were maintained at 37 °C in a humidified atmosphere containing 5% CO₂.