Fig. 7

The excessive telomerase or TRF2 abrogates the inhibitory effect of MEG3 on the growth of human liver cancer stem cells. A MEG3 was by reverse RT-PCR, and TERT was detected by Western blotting. β-actin was used as an internal reference gene. B CCK8 assay for cell proliferation capacity. C Colony formation ability assay. C (a) Photograph of colonies. C (b) Analysis of cell colony formation rate. D The formation rate of sphere was measured. F Tumorigenesis test in vivo. F (a) Photographs of transplanted tumors (xenograft). F (b, c) 4% formalin-fixed, paraffin-embedded transplanted tumor tissue sections (4 μm) were subjected to immunohistochemical staining for anti-PCNA. Comparison of PCNA positive rates of transplanted tumors. E MEG3 was detected by RT-PCR, and the expression of TRF2 was detected by Western blotting. β-actin was used as an internal reference gene. F Cell proliferation ability was determined by CCK8 method. G (a) Photograph of plate colonies. G (b) The analysis of colony formation rate. H The assay for sphere formation ability. I (a) photograph of a transplanted tumor (xenograft). I (b) The comparison of xenograft tumors size (g). I (c) The comparison of xenograft tumor appearance time. I (d) Comparison of PCNA positive rates. J Schematic diagram of the molecular mechanism of MEG3 affecting the growth of human liver cancer stem cells