Siakt

The human (BCa) cell lines MDA-MB-453 and MDA-MB-231 were obtained from American Type Culture Collection (Rockville, MD, USA) and cultured following the manufacturer’s recommendations. MDA-MB-453 and MDA-MB-231 cells are negative for nuclear progesterone receptor (nPR). The cell counting kit-8 (CCK8) was purchased from BestBio Biotechnology (Shanghai, China).
The plasmid pEGFP/C-BCRP harboring full-length BCRP cDNA was constructed and stored in our laboratory; this plasmid was transfected into MDA-MB-453 and MDA-MB-231 cells to establish the MDA-MB-453/BCRP and MDA-MB-231/BCRP stable cell lines as previously reported (11 (link)). The siAkt was purchased from GenePharma (Shanghai, China) and Akt inhibitor of perifosine was obtained from MedChemExpress (Shanghai, China).

PCR production of PTEN ampliation was cloned into pmirGLO vector (Promega). MiR-20a mimic, inhibitor and miR-NC were synthesized by GenePharma Co.Ltd. (Suzhou, China). LncRNA PTENP1 pcDNA3.1 vector (PTENP1, Invitrogen, CA, USA), LV-NC, LV-PTENP1, siPTENP1, shPTENP1, siAkt, siSCR and shSCR were obtained from GenePharma Co.Ltd. (Suzhou, China). The transfection assay was conducted with Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) for incubation. The transfected efficiency was measured by qRT-PCR.