Fig. 2

NEAT1 enhances invasive and sphere-forming capabilities and confers paclitaxel chemoresistance in aggressive EC cells. (a) RNA sequencing was used to measure the expression levels of one NEAT1 isoform across 545 TCGA EC patient samples and two PVT1 isoforms across 290 TCGA kidney cancer patient samples obtained from the ISOexpresso platform. Each color represents a specific isoform. (b) qRT-PCR was used to analyze NEAT1 expression in the normal endometrial epithelial cell line EM, the serous EC cell line SPAC-1-L, invasive HI cells, sphere-forming HEC-50 cells, TX-resistant HEC-50 cells and parental HEC-50 cells. (c) The efficiency of NEAT1 expression knockdown in SPAC-1-L and HI cells by two shRNAs was verified using qRT-PCR analysis. (d-f) The effects of NEAT1 expression knockdown on cell colony formation (d), cell invasion (e) and sphere formation (f) were examined using colony formation, Matrigel invasion and sphere formation assays, respectively, in SPAC-1-L and HI cells. (g, h) Cell survival was examined by a cell viability assay in SPAC-1-L (g) and HI (h) cells transfected with or without NEAT1-specific shRNAs and treated with TX. (i) Representative images from colony formation, invasion and sphere formation assays using HI cells transfected with or without NEAT1-specific shRNAs are shown. (j) A heat map of qRT-PCR data shows the expression of the indicated genes in SPAC-1-L and HI cells transfected with or without NEAT1-specific shRNAs. (K, L) Growth curves (k) and quantification of the weight (l) of subcutaneous control or NEAT1-silenced SPAC-1-L xenografts treated for 24 days with vehicle or TX are shown. (m) Representative IHC images of Ki-67 expression in tumors from the mice described in K and L are shown. Scale bar: 50 μm. *P < 0.05