Fig. 2
Ectopic expression of SNHG1 alone was sufficient to drive to the malignant transformation of normal human bladder urothelial cell and to enhance the anchorageindependent growth of human BMIBC cell lines. A, C, E UROtsa (A), U5637 (C) and T24T (E) cells were stably transfected with a plasmid constitutively expressing human SNHG1 and real-time PCR was performed to identify the stable transfectants. B, D, F UROtsa(SNHG1) and UROtsa(Vector) (B), U5637(SNHG1) and U5637(Vector) (D), T24T(SNHG1) and T24T(Vector) (F) cells underwent an anchorage-independent assay, the number of colonies was counted, and the results were presented as colonies per 50,000 or 10,000 cells, with bars representing the mean ± SD. In (B), An asterisk (*) indicates a significant increase compared to the medium control in UROtsa(Vector) cells, and the symbol (#) denotes a significant increase compared to UROtsa(Vector) cells treated with EGF (p < 0.01). In (A, C-F), an asterisk (*) signifies a significant increase compared to cells transfected with Vector. G and I Cell extracts from various U5637 (G) or T24T (I) transfectants were analyzed by real-time PCR to identify the knockdown efficiency of shRNA targeting SNHG1. H and J U5637(shSNHG1#1) cells, U5637(shSNHG1#2) cells versus U5637(Nonsense) cells (H), T24T(shSNHG1#1) cells, T24T(shSNHG1#2) cells versus T24T(Nonsense) cells (J) were subjected to an anchorage-independent soft agar growth assay as detailed in the Materials and Methods section. The number of colonies, each with more than 32 cells, was counted, and the results were presented as colonies/104 cells. Bars show the mean ± SD, and an asterisk (*) signifies a significant decrease compared to UROtsa(Nonsense) transfectants or T24T(Nonsense) transfectants (p < 0.01)