Figure 2
CMA promotes breast cancer cell growth and survival. (A) The stable inhibitory efficiency of shRNAs against LAMP2A in MDA-MB-231 and MDA-MB-468 cells was detected. (B) Western blotting detected LAMP2A overexpression by transient transfection in two breast cancer cell lines. (C) The LAMP2B mRNA level was determined in MDA-MB-231 and MDA-MB-468 cells by qRT-PCR. (D) Purified GAPDH protein was incubated with intact lysosomes isolated from the stable LAMP2A knockdown and overexpression cells and then harvested, fractionated and immunoblotted with a GAPDH antibody. (E) The cell growth rate was determined by an MTT assay at indicated timepoints in MDA-MB-231 cells and MDA-MB-468 cells. (F) A colony formation assay was performed to detect the proliferative capability of the breast cancer cells. The cells were seeded onto 6-well plates and allowed to form colonies for two weeks. All values are expressed as the mean ± SD of three different experiments; *P < 0.05 and **P < 0.01, shLAMP2A versus shControl at the same timepoint; # P < 0.05, LAMP2A overexpression group versus the control group at the same timepoint. (G) 1 × 107 shLAMP2A or shControl MDA-MB-231 cells were subcutaneously injected into nude mice. The LAMP2A protein levels in the tumors at the time of resection were determined by immunoblots (left), and the size of the tumors was monitored by the standard formula length × width × width × 0.5 (right) (n = 5; *P < 0.05, shLAMP2A or shControl).
