Caski

Human colon cancer cell lines: SW 948 (ATCC, no. CCL-237) and SW620 (ATCC, no. CCL-227), human osteosarcoma cell lines: Saos-2 (ATCC, HTB-85) and HOS (ATCC, CRL-1543), human cervical cancer cell lines: SiHa (ATCC, HTB-35) and CaSki (ATCC, CRM-CRL-1550), human normal cells: epithelial cells of the large intestine (CoTr): CCD 841 CoTr (ATCC, CRL-1807), and osteoblasts (hFOB): hFOB 1.19 (ATCC, CRL-11372) were used in these studies. Both normal cell lines were transformed with thermo-labile virus SV-40. All cell lines, purchased from Sigma-Aldrich (St. Louis, MO, USA), were maintained in appropriate media recommended by ATCC, namely, colon cancer cell lines: L-15 medium; Saos-2 cell line: McCoy’s 5A Modified Medium; HOS and SiHa cell lines: MEM Medium; CaSki cell line: RPMI medium; hFOB cell line: 1:1 mixture of DMEM without phenol red and Ham’s F12 medium, and CoTr: DMEM Medium. All media were supplemented with fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μg/mL streptomycin (Sigma Aldrich), except the medium for hFOB, which contained 0.3 mg/mL of G418. The cell cultures were cultivated in standard conditions at 37 °C, 95% humidity, and with 5% CO₂, except the SW 620 cell line cultivated without CO_2, and hFOB and CoTr cultivated at a permissive temperature of 34 °C.

A panel of cervical cancer cell lines including SiHa and CaSki was purchased from ATCC (Supplementary Table S1). SiHa (PIK3CA-wt) and CaSki (PIK3CA-wt/PIK3CA-E545K) cell lines were cultured in MEM and RPMI (Invitrogen), respectively, supplemented with 10% (v/v) fetal bovine serum (FBS, Hyclone III, Invitrogen) and penicillin (50 units/ml) and streptomycin (50 μg/ml). HeLa cells (PIK3CA-WT) were cultured in DMEM with 5% (v/v) serum and antibiotics as above. Stable cell lines derived from parental HeLa cell line (described below and in Supplementary Material) were cultured in DMEM supplemented with 5% fetal bovine serum, streptomycin and penicillin as above and other antibiotics as indicated below. All cell lines were cultured in a humidified incubator under an atmosphere of 5% CO₂ at 37°C. SiHa, CaSki and HeLa cell lines were authenticated by STR analysis by ATCC January 2016.

Human CC cell lines (SiHa, CaSki, ME-180, C4-1) and human normal cervical cell line (Ect1/E6E7), from ATCC (Rockville, Maryland), were allowed to grow under 37 °C and 5% CO₂ in the DMEM (Invitrogen, Carlsbad, CA). The 1% antibiotics and 10% FBS, both from Invitrogen, were acquired for purpose of cell culture. Besides, 20 mmol/l of LiCl, 10 mM of DMSO and 20 nM of C646 were all purchased from Sigma Aldrich (St. Louis, MI) to treat SiHa and CaSki cells.

The human embryonic kidney cell line HEK293, breast cell lines (HBL100, MCF-7, and MDA-MB-231) and cervical cancer cell lines (HeLa, CaSki, C-33A, and SiHa) were purchased from ATCC (Manassas, VA, USA). Cells were cultured in DMEM or RPMI 1640 supplemented with 10% fetal bovine serum, 100 U/ml penicillin, 100 μg/ml streptomycin, and 2 mM L-glutamine at 37 °C in a 5% CO₂ incubator. Transfection was carried out using Lipofectamine 2000 according to the manufacturer’s instructions (Invitrogen, Carlsbad, CA, USA).