Fig. 2. PVT1 knockdown inhibited the stemness and TMZ resistance of glioma cells. (A) The expression level of PVT1 determined by RT-qPCR, was significantly up-regulated in glioma cells T98G, A172, U87, U251 and U373 compared with that in human astrocyte normal cells HEB. GAPDH was used for normalization. (B) The expression level of PVT1 in U87 and U251 cells after transfected with PTV1 shRNA and NC shRNA were detected by RT-qPCR. GAPDH was used for normalization. The results revealed that PVT1-shRNA expression vector was successfully constructed. (C) Cell proliferation was assessed by CCK8 and the results of CCK8 revealed that PVT1 knockdown inhibited glioma cell proliferation. (D) PVT1 knockdown inhibited glioma cell migration, as indicated by wound healing assay. (E) PVT1 knockdown inhibited glioma cell invasion, as indicated by transwell assay. (F) The rate of CD133+ U87 and U251 cells was suppressed by PVT1 knockdown, as indicated by flow cytometry assay. (G) The protein level of ELF4 and stemness factors (SOX2, Oct4, Nanog) in glioma cells were decreased after PVT1 knockdown, as indicated by western blot. GAPDH was used for normalization. (H) PVT1 knockdown inhibited resistance of U87 and U251 cells to TMZ, as detected by CCK8 assay. Data are presented as Mean±SD of three independent experiments. *p<0.05, **p<0.01, ***p<0.001.
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