Caski

Twenty-four cases of cervical carcinoma samples and paratumor tissues were collected from Binzhou Central Hospital. Written informed consent for participation was obtained from patients before participation in this study. The ethical approval was obtained from the ethics committee of the Binzhou Central Hospital. The research conforms to the Code of Ethics of the World Medical Association (Declaration of Helsinki) printed in the British Medical Journal (July 18, 1964). The cervical cancer cell lines, including C-33 A, C-4-I, SiHa and CaSki, were purchased from the Chinese Academy of Sciences Cell Bank of Type Culture Collection (CBTCCCAS, Shanghai, People’s Republic of China). The human cervical surface epithelial cell, HcerEpic was obtained from Nanjing Cobioer (Nanjing, Jiangsu, People’s Republic of China). The HEK-293T cell was purchased from GuangZhou Jennio (Guangzhou, Guangdong, People’s Republic of China). The SiHa and CaSki cells were cultured in 1,640 media supplemented with 10% FBS (Thermo Fisher Scientific, Carlsbad, CA, USA), 100 μg/mL streptomycin/penicillin. The C-33 A, C-4-I, HcerEpic and HEK-293T cells were cultured using DMEM media containing 10% FBS (Thermo Fisher Scientific). Cells were cultured in an incubator containing 95% air and 5% CO₂ at 37°C.

Human cervical squamous cell carcinoma cell lines (Ca-Ski, Hela, SiHa and c-33A) were obtained from Shanghai Institute of Biochemistry and Cell Biology (Shanghai, China). A non-cancerous ectocervical epithelial cell line (Ect1/E6E7) was obtained from American Type Culture Collection (ATCC) and maintained in Gibco^® Keratinocyte-Serum Free medium with 0.1 ng/ml human recombinant epidermal growth factor, 0.05 mg/ml bovine pituitary extract, and 0.4 mM calcium chloride. Ca-Ski cells were cultured in the Gibco^® RPMI-1640 medium and the other cells were cultured in the Gibco^® DMEM medium (Thermo Fisher Scientific, Inc.) supplemented with 10% Gibco FBS (Thermo Fisher Scientific, Inc.) and 1% penicillin-streptomycin. All cell lines were incubated at 37˚C in an incubator in an atmosphere containing 5% CO₂. The cells were treated with 30 µM Tat-BECN1 (Selleck Chemicals), a potent and specific autophagy inducer via activating Beclin1 (19 ,20 (link)), or 200 ng/ml TNF-related apoptosis-inducing ligand (TRAIL; Merck KGaA).

The immortalized human cervical epithelial cell line H8 and four human CC cell lines (Caski, ME180, C-33A and Siha) were all bought from BeNa Culture Collection (Suzhou, Jiangsu, China). Dulbecco’s Modified Eagle Medium (DMEM), purchased from Sigma (St Louis, MO, USA) was used to culture the H8, C-33A and Siha cell lines. Roswell Park Memorial Institute 1640 (RPMI 1640) medium, purchased from Thermo Fisher Scientific (Waltham, MA, USA), was applied to culture Caski cell lines. McCOY’s 5A medium, purchased from Thermo Fisher Scientific, was used to culture ME180 cell lines. 10% Fetal bovine serum (FBS), streptomycin (100 mg/mL) and penicillin (100 unit/mL), all purchased from Gibco (Waltham, MA, USA), were supplemented into the culture medium. And the cell lines used in the present study were all cultured in a incubator (containing 5% CO₂) maintained at 37°C.