Hec59

The three human cancer cell lines (HEC59, JHUM3, and AN3CA) were purchased from the Japanese Collection of Research Bioresources Cell Bank, RIKEN Cell Bank, and American Type Culture Collection, respectively. All cell lines were cultured in Dulbecco’s modified Eagle medium, supplemented with 10% fetal bovine serum, 50 IU/mL penicillin, and 50 μg/mL streptomycin.
Tissues from 122 Japanese patients who were diagnosed with endometrial cancer between 2006 and 2014 at Niigata University Medical and Dental Hospital were included in this study. Fresh frozen samples were obtained from primary tumor tissues during surgery prior to chemotherapy. The histologic characteristics of surgically resected specimens were assessed on formalin-fixed and paraffin-embedded hematoxylin and eosin sections by a gynecologic pathologist. A total of 122 samples (109 endometrioid adenocarcinomas, 5 mixed adenocarcinomas, 4 carcinosarcomas, 2 undifferentiated adenocarcinomas, 1 serous adenocarcinoma and 1 unclassified adenocarcinoma of the uterine corpus) were analyzed. The institutional ethics review board at Niigata University approved this study. All patients provided written informed consent for the collection of samples and subsequent analysis. All experiments were performed in accordance with the approved guidelines.

AN3CA (ATCC, HTB-111) and HEC-1A (ATCC, HTB-112) cells were obtained from the American Type Culture Collection. HEC-59 (JCRB1120) and Ishikawa (JCRB1505) cells were purchased from the Japanese Collection of Research Bioresources (JCRB) cell bank. AN3CA cells were cultured and grown in Minimum Essential Medium (MEM) (GIBCO, 41500-034) supplemented with 10% fetal bovine serum (GIBCO, 10437). HEC-59 and Ishikawa cells were cultured in Minimum Essential Medium (MEM) (GIBCO, 41500-034) supplemented with 15% fetal bovine serum. HEC-1A cells were cultured in McCoy’s 5A Medium (SIGMA, M4892) supplemented with 10% fetal bovine serum. All cells were cultured at 37 °C in a humidified atmosphere of 5% CO_2. PQR309 (Bimiralisib; 23441) and KJ-Pyr-9 (19116) were purchased from Cayman Chemical (Ann Arbor, MI). Chloroquine diphosphate salt (C 6628) was purchased from Sigma (St. Louis, MO, USA).