Shg44

Human glioma cell lines (U251, U87, SHG‐44, and SHG139) and the normal human astrocytes cell line (NHA) were purchased from Cell Bank of Chinese Academy of Sciences (Shanghai, China) and cultured in DMEM or RPMI 1640 medium containing 10% fetal bovine serum (Gibco, Carlsbad, CA, USA), at 37°C with 5% CO2. All media were additionally supplemented with 100 U/mL penicillin and 100 mg/mL streptomycin (Gibco, Invitrogen, Shanghai, China).

Human glioma cell lines, LN118, SHG44, U87, U251, and A172, were obtained from Cell Bank of Chinese Academy of Sciences (Pudong, Shanghai, China). The cells were grown in the Roswell Park Memorial Institute (RPMI) 1640 medium, supplemented with 10% fetal bovine serum (FBS) (Yunshan Technology, Haidian, Beijing, China), 100 U/ml penicillin G (Yunshan Technology, Haidian, Beijing, China), and 100 μg/ml streptomycin (Sigma, Suzhou, Jiangsu, China). Normal human astrocytes (NHA) were obtained from Weihui Biot (Hangzhou, Zhejiang, China) and were grown in Dulbecco's Modified Eagle's Medium with high glucose and sodium pyruvate.
The siRNAs used to knockdown ZNF667-AS1 were purchased from GEMA GENE (Pudong, Shanghai, China). The siRNA sequences were as follows: si-ZNF667-AS1: CTACCACAGCTTCCATG; si-ZNF667-AS1-2: GCCCACTGTATTCAACA. Cell transfection was conducted by using the LipofectamineTM 2000 transfection reagent (Invitrogen, Guangzhou, Guandong, China) according to the manufacturer's instructions. After transfection for 24 h, the knockdown efficiency was evaluated by RT-PCR.

The human glioma cell lines U251, LN229, T98G, A172, and SHG44 were obtained from the Cell Bank of Chinese Academy of Sciences (Shanghai, China). Normal Human Astrocyte (NHA) cells were purchased from the American Type Cell Culture Collection (ATCC; Rockville, MD, USA). All cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; HyClone; GE Healthcare Life Sciences, Logan, UT, USA) with 10% fetal-bovine serum (Gibco; Thermo Fisher Scientific, Inc., Los Angeles CA, USA) in a 37 °C, 5% CO₂ humidified atmosphere.
GBM samples and adjacent normal brain tissues were collected from 50 patients who underwent tumor resection at Harbin Medical University Cancer Hospital between December 2012 and December 2016 Additional file 3: Table. S1. GBM patients had not received radiotherapy or chemotherapy treatment prior to surgery. All tissue samples were pathologically confirmed and immediately snap-frozen in liquid nitrogen until RNA extraction. Written informed consent to the use of the tissue samples for research purposes was obtained from each patient. All procedures were conducted in accordance with the principles outlined in the Declaration of Helsinki, and all applicable international, national and/or institutional guidelines for the care and use of animals were followed. The study protocol was approved by the Ethics Committee of Harbin Medical University (Harbin, China).

The human glioma cell line of SHG-44 was obtained from Cell Bank of Chinese Academy of Sciences (Shanghai, China). Cells was cultured in DMEM supplemented with 10% (v/v) FBS and antibiotics (50 U/ml penicillin and 100 µg/ml streptomycin, Invitrogen) at 37°C in 5% CO₂. WZY-321 was constructed by our lab and its chemical structure was shown in Fig. 1A.

We purchased human cortical astrocyte cell line, HA1800, human glioma cell lines (T98G, SHG44, U87MG, and U251MG), and human umbilical vein endothelial cells (HUVECs) from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). The cells were cultured in DMEM medium (Thermo Fisher Scientific) containing 10% FBS and 100 U/mL penicillin/streptomycin in a humidified incubator at 37° C and 5% CO₂.

The glioma cell lines SHG-44 and U87 were purchased from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). All cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 50 IU/ml penicillin and 50 mg/ml streptomycin (Gibco, Grand Island, NY, United States) under the conditions of 37°C in a humidified atmosphere with 5% CO₂.