E3 Ubiquitin Ligase CHIP Inhibits the Interaction between Hsp90β and MAST1 to Repress Radiation Resistance in Non-Small-Cell Lung Cancer Stem Cells

Figure 3

Overexpression of CHIP inhibits the NSCLC stem cell properties and radiation resistance. (a) RT-qPCR analysis for CHIP expression in the CD133⁺ NSCLC and A549 cells treated with lentiviral oe-CHIP. (b) RT-qPCR analysis for the expression of stemness-related transcription factors (Oct4, SOX2, and Nanog) in CD133⁺ A549 cells in response to CHIP overexpression alone or combined with 4 Gy radiation. (c) Sphere formation analysis after 7 days of sphere culture in CD133⁺ A549 cells in response to CHIP overexpression alone or combined with 4 Gy radiation. (d) Cell survival analyzed by colony formation assay in CD133⁺ A549 cells exposed to different doses of radiation after 9 days of culture. (e) Annexin V/PI double staining for detection of apoptosis of CD133⁺ A549 cells in response to CHIP overexpression alone or combined with 4 Gy radiation. (f) Immunofluorescence detection of the number of γ-H2AX foci in CD133⁺ A549 cells in response to CHIP overexpression alone or combined with 4 Gy radiation. (g) The quantitation of tumor volume in nude mice subcutaneously injected with CD133⁺ A549 cells in response to CHIP overexpression alone or combined with 4 Gy radiation (). (h) Statistics of tumor weight of nude mice in response to CHIP overexpression alone or combined with 4 Gy radiation (). . Cell experiments were independently repeated three times.

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)